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Patent Analysis of

Use of the chromosome 19 microRNA cluster (C19MC) for treating microbial disease and promoting authophagy

Updated Time 12 June 2019

Patent Registration Data

Publication Number

US10000755

Application Number

US15/417717

Application Date

27 January 2017

Publication Date

19 June 2018

Current Assignee

UNIVERSITY OF PITTSBURGH - OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION

Original Assignee (Applicant)

UNIVERSITY OF PITTSBURGH - OF THE COMMONWEALTH SYSTEM OF HIGHER EDUCATION

International Classification

C07H21/02,C12N15/113,C07H21/04,A61K48/00

Cooperative Classification

C12N15/1131,C12N15/1132,C12N15/1133,C12N2310/141,C12N15/113

Inventor

SADOVSKY, YOEL,CANDRILLI, CAROLYN COYNE

Patent Images

This patent contains figures and images illustrating the invention and its embodiment.

US10000755 Use chromosome 19 1 US10000755 Use chromosome 19 2 US10000755 Use chromosome 19 3
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Abstract

It is disclosed herein that cultured primary placental human trophoblast (PHT) cells are highly resistant to infection by a number of disparate viruses, and confer this resistance to non-placental recipient cells by exosome-mediated delivery of microRNAs (miRs). PHT cells express high levels of unique, primate-specific miRNAs, expressed from the chromosome 19 miRNA cluster (C19MC). It is further disclosed herein that C19MC miRNAs are packaged within PHT-derived exosomes and attenuate viral replication in recipient cells by inducing autophagy. Thus, provided herein are methods of inhibiting, treating or preventing microbial infections by administering one or more miRs of the C19MC. Also provided are methods of inducing autophagy in a cell by contacting the cell with one or more miRs of the C19MC.

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Claims

1. A method of inducing autophagy in a cell, comprising contacting the cell with an effective amount of one or more miRs encoded by the C19MC, wherein the one or more miRs comprise miR-517-3p, miR-516b-5p, miR-512-3p, or any combination thereof, thereby inducing autophagy in the cell.

2. The method of claim 1, wherein the one or more miRs comprises miR-517-3p, miR-516b-5p, and miR-512-3p.

3. The method of claim 1, wherein the one or more miRs comprises miR-517-3p, miR-1323, miR-516b-5p, miR-525-5p, miR-512-3p and miR-515-3p.

4. The method of claim 1, wherein the method is an in vitro method.

5. The method of claim 4, wherein contacting the cell with the one or more miRs encoded by the C19MC comprises transfecting a nucleic acid molecule encoding the C19MC or a biologically active portion thereof.

6. The method of claim 5, wherein the nucleic acid molecule comprises a vector.

7. The method of claim 1, wherein the method is an in vivo method and contacting the cell comprises administering to a subject an effective amount of the one or more inhibitory miRs encoded by the C19MC.

8. The method of claim 7, wherein administering the one or more inhibitory miRs to the subject comprises administering a nucleic acid molecule encoding the C19MC or a biologically active portion thereof.

9. The method of claim 8, wherein the nucleic acid molecule comprises a vector.

10. The method of claim 7, wherein the subject suffers from a disease associated with a deficiency in autophagy.

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Claim Tree

  • 1
    1. A method of inducing autophagy in a cell, comprising
    • contacting the cell with an effective amount of one or more miRs encoded by the C19MC, wherein the one or more miRs comprise miR-517-3p, miR-516b-5p, miR-512-3p, or any combination thereof, thereby inducing autophagy in the cell.
    • 2. The method of claim 1, wherein
      • the one or more miRs comprises
    • 3. The method of claim 1, wherein
      • the one or more miRs comprises
    • 4. The method of claim 1, wherein
      • the method is an in vitro method.
    • 7. The method of claim 1, wherein
      • the method is an in vivo method and contacting the cell comprises
See all independent claims <>

Description

FIELD

This disclosure concerns the use of microRNAs encoded by the primate-specific chromosome 19 miRNA cluster (C19MC) for the prophylaxis and/or treatment of microbial infection and diseases associated with autophagy.

BACKGROUND

Strategies to reduce the risk of fetal infection are of critical importance during pregnancy, where maternal to fetal transmission of microbes can have devastating consequences to the developing embryo, ranging from fetal infection, induced preterm delivery, structural or functional congenital anomalies, miscarriages and stillbirths (Ornoy and Tenenbaum, Reprod Toxicol 21, 446-457, 2006; Silingardi et al., Am J Forensic Med Pathol 30, 394-397, 2009; Euscher et al., Obstet Gynecol 98, 1019-1026, 2001). Additionally, pathogenic infections can compromise maternal health and jeopardize the pregnancy even in the absence of fetal transmission. The physical barrier interfacing the maternal and fetal blood systems within human hemochorial placenta villi include the trophoblast bilayer, basement membrane, stromal cells and fetal capillary endothelial cells. The multinucleated, terminally differentiated villous syncytiotrophoblasts are bathed directly in the maternal blood, and mediate the crucial exchange of gases, nutrients, and waste products between the mother and fetus, produce crucial hormones, and immunologically guard the developing fetus. These cells, along with the less differentiated cytotrophoblasts, constitute the first line of feto-placental defense against invading microbes.

Intrauterine transmission of viruses is likely to occur by at least four potential routes: (a) transmission across the placental villous trophoblasts by hematogenous spread or ascending infection, (b) placental transfer of infected macrophages from the maternal blood, (c) transfer of viruses via paracellular routes and/or (d) transmission of viruses from the infected maternal endothelial microvasculature to endovascular extravillous cytotrophoblasts. In general, little is known regarding the defense mechanisms employed by placental trophoblasts to defend against viral infections. Additionally, as antiviral therapeutics are generally ineffective in preventing intrauterine viral infections, elucidating the nature of these mechanism(s), as well as the underpinnings of viral counter-measures, is critical for designing therapeutic strategies aimed at preventing fetal and maternal viral disease.

Mammalian cells utilize diverse defense mechanisms to combat microbial pathogens. One crucial mechanism is the induction of autophagy, an evolutionarily conserved lysosomal degradation pathway that has been associated with an array of cellular functions, including cell death (Beaulation and Lockshin, J Morphol 154:39-57, 1977; Liang et al., Nature 402:672-676, 1999), tumorigenesis (Qu et al., J Clin Invest 112:1809-1820, 2003), and neurodegeneration (Hara et al., Nature 441:885-889, 2006; Komatsu et al., Nature 441:880-884, 2006). Autophagy also degrades intracellular foreign microbial invaders (a process sometimes referred to as xenophagy or virophagy). The cascade of events that culminate in autophagy begin with the formation of a double membrane organelle, the autophagosome, and ends in the degradation of engulfed material via the fusion of autophagosomes with late endosomes and/or lysosomes. The degradation of microbes via the fusion of autophagosomes with lysosomes is a key component in the antimicrobial effects of autophagy, yet the sequestration of viruses into autophagosomes can also direct MHC class II presentation (English et al., Nat Immunol 10:480-487, 2009), the production of antiviral type I interferons downstream of toll-like receptor 7 engagement (Lee et al., Science 315:1398-1401, 2007), and even altered T-cell signaling (Nedjic et al., Nature 455:396-400, 2008). It is becoming clear that autophagy functions at the crossroads of many aspects of cell survival, and is likely a fundamental component of antiviral signaling.

SUMMARY

It is disclosed herein that microRNAs (miRs) of the C19MC cluster promote viral resistance and induce autophagy of recipient cells.

Provided herein is a method of inhibiting or treating a microbial infection in a subject by selecting a subject with a microbial infection or at risk for contracting a microbial infection; and administering to the subject a therapeutically effective amount of one or more miRs encoded by the C19MC, thereby inhibiting or treating the microbial infection. In some embodiments, the one or more miRs are administered by administering a nucleic acid molecule encoding the entire C19MC or a biologically active portion thereof, for example a portion that encodes one or more miRs that promote viral resistance and induce autophagy. In non-limiting examples, the one or more miRs include miR-517-3p, miR-1323, miR-516b-5p, miR-525-5p, miR-512-3p, or miR-515-3p, or any combination thereof.

Also provided herein is a method of inducing autophagy in a cell by contacting the cell with an effective amount of one or more miRs encoded by the C19MC, thereby inducing autophagy in the cell. In some embodiments, the method is an in vitro method. In other embodiments, the method is an in vivo method that includes administering to a subject an effective amount of one or more miRs encoded by the C19MC, or a nucleic acid molecule encoding the C19MC or a biologically active portion thereof. In non-limiting examples, the subject suffers from a disease associated with a deficiency in autophagy and/or a disease that may be ameliorated by enhancing autophagy.

The foregoing and other objects, features, and advantages of the invention will become more apparent from the following detailed description, which proceeds with reference to the accompanying figures.

BRIEF DESCRIPTION OF THE DRAWINGS

FIGS. 1A-1E: Conditioned primary human trophoblast (PHT) medium and exosomes confer viral resistance to recipient cells. (FIG. 1A) PHT or non-PHT cells were infected with a panel of viruses, including coxsackievirus B (CVB), poliovirus (PV), vesicular stomatitis virus (VSV), vaccinia virus (VV), herpes simplex virus-1 (HSV-1), or cytomegalovirus (CMV). Non-PHT cells were as follows: HeLa (CVB, PV), U2OS (VSV, HSV-1, and VV), and human foreskin fibroblasts (HFF, CMV). Shown are the percent infected cells (assessed by immunofluorescence (IF); *p<0.0001). (FIG. 1B) Non-PHT recipient cells were exposed for 24 h to non-conditioned or conditioned PHT medium, and then infected with CVB, VSV, HCV, or VV. Non-PHT cells were as follows: HFF (CVB), U2OS (VSV, VV), and Huh 7.5 (HCV). Shown are the percent of infected cells, assessed by IF (CVB, VSV), luciferase assay (HCV), or RT-qPCR (VV); *p<0.05, **p<0.005. (FIG. 1C, left) Cells were exposed to non-conditioned or conditioned PHT medium for 24 h, then infected with VSV or CVB. (FIG. 1C, right) Primary cells were infected with VSV following exposure to non-conditioned or conditioned PHT medium (*p<0.05, **p<0.005). (FIG. 1D) Conditioned PHT medium was subjected to heat inactivation or sonication prior to 24 h exposure to Vero cells, then infected with VSV. Percent infection assessed as in (A); (*p<0.0001). (FIG. 1E) U2OS cells were exposed for 24 h to non-conditioned, conditioned, exosome-depleted conditioned medium, exosomes purified from PHT, JEG-3, or from three preparations of murine dendritic cell (DC), and then infected with VSV. Percent infection assessed as in (A); (*p<0.0005), each PHT exosome preparation was derived from a different placental preparation.

FIGS. 2A-2E: PHT and exosomal C19MC miRNAs confer viral resistance to recipient cells. (FIG. 2A) U2OS cells stably expressing control- or C19MC-bacterial artificial chromosome (BAC) were infected with VSV (infection levels assessed by RT-qPCR, *p<0.0001). (FIG. 2B) U2OS cells were transfected with C19MC miRNA mimics that represent the miRNA sub-groups detailed in Table 2 or control mimics, and then infected with VSV (shown as percent infected cells, assessed by IF; *p<0.05, **p<0.001). (FIG. 2C) U2OS cells, transfected with mimics of the six highest expressed C19MC miRNAs, scrambled control, or non-C19MC (miR-1, -424, -630, -720) miRNA mimics, were infected with VSV (infection level assessed by IF or qPCR; *p<0.0005). (FIG. 2D) U2OS cells, transfected with mimics of the top three antiviral C19MC miRNAs or with scrambled control mimics, were infected with VSV (infection assessed by RT-qPCR; *p<0.05, **p<0.0001). (FIG. 2E) U2OS cells, transfected with scrambled control or miR-517-3p mimic, were infected with VV or HSV-1; infection assessed as in (D) (*p<0.0001).

FIGS. 3A-3D: PHT-derived exosomes induce autophagy in recipient cells. (FIG. 3A) U2OS cells transfected with mRFP-LC3b were exposed to non-conditioned-, PHT conditioned-, exosome-depleted conditioned PHT medium, or purified PHT exosomes for 24 h, and LC3b punctae formation was assessed by confocal microscopy. Shown are confocal micrographs (top) and quantification of mRFP-LC3b punctae per cell (bottom) (*p<0.0001). (FIG. 3B, top) Electron micrographs of cells exposed to non-conditioned or conditioned PHT medium (Vero), exosome-depleted conditioned PHT medium (Vero), or purified PHT exosomes (U2OS). Arrows denote autophagosomes. Bar=500 nm. (FIG. 3B, bottom) Quantification of electron micrographs of cells exposed to non-conditioned (Vero and U2OS), conditioned PHT media samples (Vero and U2OS), exosome-depleted conditioned medium (Vero), or purified PHT exosomes (U20S) (*p<0.0001). (FIG. 3C) U2OS cells transfected with mRFP-LC3b were exposed to non-conditioned or conditioned PHT medium in the absence or presence of 3-methyladenine (3-MA) for 8 h, and LC3b punctae formation was assessed by confocal microscopy. Shown are confocal micrographs (top) and quantification of mRFP-LC3b punctae (bottom) (*p<0.0005). (FIG. 3D, top) Immunofluorescence images of VSV entry into U2OS cells transiently transfected with mRFP-LC3b exposed to non-conditioned (left) or conditioned (right) PHT medium (inset, 5× magnification). (FIG. 3D, bottom) Quantification of the extent of colocalization between VSV particles and mRFP-LC3B positive punctae (*p<0.0001).

FIGS. 4A-4D: C19MC miRNAs induce autophagy. (FIG. 4A, top) Electron micrographs of U2OS cells transfected with scrambled control or the six highest expressed C19MC miRNA mimics (Table 2). Black arrows denote autophagosomes and/or autolysosomes. Bar=500 nm. (FIG. 4A, bottom) Quantification of electron micrographs shown at top (*p<0.005), or in PHT cells. (FIG. 4B) U2OS cells were transfected with mRFP-LC3b and either scrambled control or the six highest expressed C19MC miRNA mimics. Shown are confocal micrographs (top) and quantification of mRFP-LC3b punctae per cell (bottom) (*p=0.0005). (FIG. 4C, left) Electron micrographs of U2OS cells transfected with scrambled control or the most potent antiviral miRNA mimics. Black arrows denote autophagosomes and/or autolysosomes. Bar=500 nm. (FIG. 4C, right) Quantification of adjacent electron micrographs (*p<0.005). (FIG. 4D, top) A representative immunoblot for p62 or GAPDH in U2OS cells stably transfected with either control Del- or C19MC-BAC. (FIG. 4D, bottom) Densitometry of p62 levels (normalized to GAPDH) from three independent immunoblots as described above (*p<0.05).

FIGS. 5A-5C: Suppression of autophagy restores C19MC-medated antiviral effects. (FIG. 5A) U2OS cells transfected with scrambled control or miRNA mimics of the six most prevalent C19MC miRNA mimics. Cells were exposed to 3-MA before and during VSV infection. Relative VSV RNA was analyzed by RT-qPCR (*p<0.0005). (FIG. 5B, top) U2OS cells stably expressing control- or C19MC-BAC transfected with scrambled control siRNA or beclin-1 siRNA for 72 h were infected with VSV, and relative infection was determined by RT-qPCR (*p<0.05, determined using ANOVA with Bonferroni correction). (FIG. 5B, bottom) Immunoblots for beclin-1 or actin in cells transfected as described above. (FIG. 5C) PHT cells were treated with 3-MA for 60 min prior to infection with green fluorescent protein (GFP)-VSV (in the presence of 3-MA). Relative VSV RNA was analyzed by RT-qPCR (*p<0.005). Data are representative of four independent experiments.

FIGS. 6A-6H: Medium from different preparations of PHT cells confers an antiviral effect on recipient cells. (FIG. 6A) Tissue culture infectious dose 50 (TCID50) assays for VSV in Vero cells pretreated for 24 h with non-conditioned medium (top, in triplicate) or three independent preparations of conditioned PHT medium (bottom). Cells were infected in the indicated dilution of virus in the presence of non-conditioned or conditioned medium for approximately 40-45 h and then stained with crystal violet. (FIG. 6B) Vero cells were exposed to non-conditioned (Non-cond) or conditioned (Cond) medium isolated from BeWo cells for 24 h and then infected with VSV. Shown is the percent of infected cells (as assessed by IF). (FIG. 6C) VSV was incubated in non-conditioned or conditioned PHT medium (in the absence of cells) for 1 h at 37° C. then a plaque assays performed. Shown are VSV titers (in pfu/mL). (FIG. 6D, left) U2OS cells were exposed to non-conditioned (Non-cond) or conditioned media (Cond) from two independent PHT preparations and infected with VSV. Relative VSV RNA was assessed by RT-qPCR (*p<0.0001). (FIG. 6D, right) Caco-2 or Vero cells were exposed to conditioned (Cond) medium isolated from four independent preparations of PHT cells for 24 h prior to infection with VSV. Shown is the percent of infected cells (as assessed by IF; *p<0.0005). (FIG. 6E) Huh7.5 cells were exposed to non-conditioned or conditioned medium isolated from four independent preparations of PHT cells for 24 h prior to infection with HCV. Shown is percent infection as assessed by luciferase assay (*p<0.005, **p<0.0005). (FIG. 6F) U2OS cells exposed to non-conditioned or conditioned PHT medium were infected with VSV or VV for approximately 6 h. Relative VSV or VV (early gene rpo35 or early gene GFP) RNA was assessed by RT-qPCR (*p<0.0001). (FIG. 6G) HFF cells were exposed to non-conditioned (Non-cond) or conditioned (Cond) PHT media for 24 h before and during infection with CMV. Shown is the percent of infected cells (assessed by IF; *p<0.05). (FIG. 6H) U2OS cells stably expressing control- or C19MC-BAC were infected with CMV, and infection levels assessed by RT-qPCR. Data are shown as fold-change over control (*p<0.0001). In all panels, data are displayed as mean±SD, and are representative of experiments performed a minimum of three times.

FIGS. 7A-7D: Medium from PHT cells induces autophagy in recipient cells. (FIG. 7A) Vero cells were transfected with mRFP-LC3b and at 24 h post-transfection were exposed for 24 h to either non-conditioned (Non-cond) or conditioned medium isolated from four independent PHT preparations. Cells were exposed to rapamycin (Rap) as a positive control. Shown are the levels of autophagic induction as determined by quantification of mRFP-LC3b positive punctae by confocal microscopy (*p<0.0001). (FIG. 7B) Vero and U2OS cells were transfected with mRFP-LC3b and then exposed to non-conditioned (Non-cond) or conditioned PHT medium (Cond) 24 h post-transfection. Cells were exposed to rapamycin (Rap) as a positive control. Shown are the levels of autophagic induction as determined by quantification of mRFP-LC3b positive punctae by confocal microscopy (*p<0.0001). (FIG. 7C) Relative mRNA levels in U2OS cells exposed to non-conditioned or conditioned PHT medium for 24 h, and analyzed using autophagy or toll-like receptor (TLR)-targeted RT-qPCR arrays. (FIG. 7D) U2OS cells stably expressing a control- or C19MC-BAC were transfected with mRFP-LC3b, fixed after 48 h, and analyzed for mRFP-LC3b punctae by confocal microscopy (*p<0.0001).

FIG. 8 is a schematic of the genomic organization of the primate-specific C19MC.

FIG. 9 is a graph showing inhibition of human immunodeficiency virus (HIV) replication by medium from PHT cells. TZM-bl cells were pre-incubated with PHT conditioned medium or control medium for 24 h prior to infection with HIV-1. Cells were then infected with serial dilutions of HIV for 48 h, and infection levels were assessed by Tat-induced luciferase reporter gene expression, in triplicate. The results represent two independent experiments.

FIG. 10 is a graph showing inhibition of HIV replication in cells transfected with control (scrambled), miR-517-3p (517), or miR-516-5p (516) mimics. TZM-bl cells were transfected for 48 h prior to infection with HIV-1. Cells were then infected with serial dilutions of HIV for 48 h, and infection levels were assessed by Tat-induced luciferase reporter gene expression, in triplicate. The results represent one independent experiment, performed in triplicate.

SEQUENCE LISTING

The nucleic and amino acid sequences listed in the accompanying sequence listing are shown using standard letter abbreviations for nucleotide bases, and three letter code for amino acids, as defined in 37 C.F.R. 1.822. Only one strand of each nucleic acid sequence is shown, but the complementary strand is understood as included by any reference to the displayed strand. The Sequence Listing is submitted as an ASCII text file, created on Jan. 23, 2017, 217 KB, which is incorporated by reference herein. In the accompanying sequence listing:

SEQ ID NOs: 1-59 are nucleotide sequences of miRNAs found in the C19MC.

SEQ ID NO: 60 is the nucleotide sequence of the human genomic DNA insert in the BAC RP11-1055017 clone containing region q13-42 of chromosome 19.

SEQ ID NOs: 61-82 are qPCR primers.

DETAILED DESCRIPTION

I. Abbreviations

BAC bacterial artificial chromosome

C19MC chromosome 19 microRNA cluster

CMV cytomegalovirus

Ct cycle threshold

CTB cholera toxin B

CVB coxsackievirus B

DAPI 4′,6-diamidino-2-phenylindole

DC dendritic cell

ELISA enzyme-linked immunosorbent assay

EM electron microscopy

FBS fetal bovine serum

GFP green fluorescent protein

hCG human chorionic gonadotropin

HCV hepatitis C virus

HFF human foreskin fibroblast

HIV human immunodeficiency virus

HSV herpes simplex virus

IF immunofluorescence

IFN interferon

ISRE interferon stimulated responsive element

3-MA 3-methyladenine

miR microRNA

miRNA microRNA

MOI multiplicity of infection

PEI polyethylenimine

PHT primary human trophoblasts

PV poliovirus

RNAi RNA interference

RT-qPCR reverse transcriptase quantitative polymerase chain reaction

TCID50 tissue culture infectious dose 50

TLR toll-like receptor

VSV vesicular stomatitis virus

VV vaccinia virus

YFP yellow fluorescent protein

II. Terms and Methods

Unless otherwise noted, technical terms are used according to conventional usage. Definitions of common terms in molecular biology may be found in Benjamin Lewin, Genes V, published by Oxford University Press, 1994 (ISBN 0-19-854287-9); Kendrew et al. (eds.), The Encyclopedia of Molecular Biology, published by Blackwell Science Ltd., 1994 (ISBN 0-632-02182-9); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8).

In order to facilitate review of the various embodiments of the disclosure, the following explanations of specific terms are provided:

Administration: To provide or give a subject an agent, such as a therapeutic agent (e.g. a nucleic acid molecule or a microRNA), by any effective route. Exemplary routes of administration include, but are not limited to, injection (such as subcutaneous, intramuscular, intradermal, intraperitoneal, intrathecal and intravenous), oral, intraductal, sublingual, rectal, transdermal, intranasal, vaginal and inhalation routes.

Autophagy: A lysosomal degradation pathway that is one of the primary mechanisms for maintaining cellular homeostasis. Autophagy, which means “to eat oneself,” is a self-cannibalization pathway that is known to be anti-microbial, functioning as a key innate immune pathway to degrade intracellular foreign microbial pathogens by a process termed antimicrobial autophagy or xenophagy. A diverse group of RNA and DNA viruses, bacteria and protozoa are sensitive to autophagy. Autophagy is also known to be deficient in a number of human diseases, such as inflammatory bowel disease, Crohn's disease, alcoholic liver disease, Parkinson's disease, Alzheimer's disease, heart disease, diabetes and obesity.

Chromosome 19 miRNA cluster (C19MC): A unique cluster of 46 primate-specific miRNA genes encoded by an approximately 100 kb region on chromosome 19 (19q13.41). The genomic organization of C19MC is shown in FIG. 8. A list of the mature miRNAs (miRs) encoded by the genes in the cluster, as well as their sequences, is provided in section V below. In the context of the present disclosure, a “biologically active portion” of the C19MC refers to any portion of the cluster (or any single miR or combination of multiple miRs encoded by the C19MC) that confers increased viral resistance and/or autophagy in a cell. In the context of the present disclosure, an “inhibitory miR” is a miR encoded by the C19MC with anti-microbial activity (and/or the ability to induce autophagy in a cell). Thus, in some examples, an inhibitory miR is a miR that increases viral resistance and/or induces autophagy in a cell. In some embodiments, the inhibitory miR does not inhibit CMV (or does not increase viral resistance to CMV).

Contacting: Placement in direct physical association; includes both in solid and liquid form. As used herein, “contacting” is used interchangeably with “exposed.” In some cases, “contacting” includes transfecting, such as transfecting a nucleic acid molecule into a cell.

Exosomes: Small (30-120 nm) endosome-derived membrane vesicles. Exosomes are enriched in miRNAs.

Isolated: An “isolated” biological component (such as a nucleic acid molecule, miRNA, protein, or cell) has been substantially separated or purified away from other biological components in the cell, blood or tissue of the organism, or the organism itself, in which the component naturally occurs, such as other chromosomal and extra-chromosomal DNA and RNA, proteins and cells. Nucleic acid molecules and proteins that have been “isolated” include those purified by standard purification methods. The term also embraces nucleic acid molecules and proteins prepared by recombinant expression in a host cell as well as chemically synthesized nucleic acid molecules and proteins.

Microbial infection: Infection by any type of microorganism, including viral, bacterial, fungal and protozoan infections. In some embodiments disclosed herein, the microbial infection is caused by any one of the following viruses, bacteria, fungi or protozoans:

Examples of viruses include, but are not limited to those in the following virus families: Retroviridae (for example, human immunodeficiency virus (HIV), human T-cell leukemia viruses; Picornaviridae (for example, poliovirus, hepatitis A virus, enteroviruses, human coxsackie viruses, rhinoviruses, echoviruses, foot-and-mouth disease virus); Caliciviridae (such as strains that cause gastroenteritis, including Norwalk virus); Togaviridae (for example, alphaviruses (including chikungunya virus, equine encephalitis viruses, Simliki Forest virus, Sindbis virus, Ross River virus, rubella viruses); Flaviridae (for example, hepatitis C virus, dengue viruses, yellow fever viruses, West Nile virus, St. Louis encephalitis virus, Japanese encephalitis virus, Powassan virus and other encephalitis viruses); Coronaviridae (for example, coronaviruses, severe acute respiratory syndrome (SARS) virus; Rhabdoviridae (for example, vesicular stomatitis viruses, rabies viruses); Filoviridae (for example, Ebola virus, Marburg virus); Paramyxoviridae (for example, parainfluenza viruses, mumps virus, measles virus, respiratory syncytial virus); Orthomyxoviridae (for example, influenza viruses); Bunyaviridae (for example, Hantaan viruses, Sin Nombre virus, Rift Valley fever virus, bunya viruses, phleboviruses and Nairo viruses); Arenaviridae (such as Lassa fever virus and other hemorrhagic fever viruses, Machupo virus, Junin virus); Reoviridae (e.g., reoviruses, orbiviurses, rotaviruses); Birnaviridae; Hepadnaviridae (hepatitis B virus); Parvoviridae (parvoviruses); Papovaviridae (papilloma viruses, polyoma viruses, BK-virus); Adenoviridae (adenoviruses); Herpesviridae (herpes simplex virus (HSV)-1 and HSV-2; cytomegalovirus; Epstein-Barr virus; varicella zoster virus; and other herpes viruses, including HSV-6); Poxviridae (variola viruses, vaccinia viruses, pox viruses); and Iridoviridae (such as African swine fever virus); Astroviridae; and unclassified viruses or agents (for example, the etiological agents of spongiform encephalopathies, the agent of delta hepatitis (thought to be a defective satellite of hepatitis B virus).

Examples of bacterial pathogens include, but are not limited to: Helicobacter pylori, Escherichia coli, Vibrio cholerae, Borelia burgdorferi, Legionella pneumophilia, Mycobacteria sps (such as. M. tuberculosis, M. avium, M. intracellulare, M. kansaii, M. gordonae), Staphylococcus aureus, Neisseria gonorrhoeae, Neisseria meningitidis, Listeria monocytogenes, Streptococcus pyogenes (Group A Streptococcus), Streptococcus agalactiae (Group B Streptococcus), Streptococcus (viridans group), Streptococcus faecalis, Streptococcus bovis, Streptococcus (anaerobic sps.), Streptococcus pneumoniae, pathogenic Campylobacter sp., Enterococcus sp., Haemophilus influenzae, Bacillus anthracis, corynebacterium diphtheriae, corynebacterium sp., Erysipelothrix rhusiopathiae, Clostridium perfringers, Clostridium tetani, Enterobacter aerogenes, Klebsiella pneumoniae, Pasturella multocida, Bacteroides sp., Fusobacterium nucleatum, Streptobacillus moniliformis, Treponema pallidium, Treponema pertenue, Leptospira, Bordetella pertussis, Shigella flexnerii, Shigella dysenteriae and Actinomyces israelli.

Examples of fungal pathogens include, but are not limited to: Cryptococcus neoformans, Histoplasma capsulatum, Coccidioides immitis, Blastomyces dermatitidis, Chlamydia trachomatis and Candida albicans.

Other pathogens (such as parasitic pathogens) include, but are not limited to: Plasmodium falciparum, Plasmodium vivax, Trypanosoma cruzi and Toxoplasma gondii.

MicroRNA: MicroRNAs (also known as miRNAs and miRs) are short RNA sequences expressed from longer transcripts found in the genomes of animals, plants and viruses and at least one single-celled eukaryote (Molnár et al., Nature 447:1126-1129, 2007; Zhao et al., Genes Dev. 21:1190-1203, 2007). MicroRNAs regulate the expression of target genes by binding to complementary sites in the target gene transcripts to cause translational repression or transcript degradation (Pillai et al., Trends Cell Biol. 17:118-126, 2007). These small RNA molecules have been implicated in a number of biological processes related to development, cell proliferation, apoptosis, metabolism, morphogenesis and diseases (Kloosterman and Plasterk, Dev. Cell 11:441-450, 2006).

A gene encoding a microRNA is transcribed to form a primary transcript microRNA (pri-miRNA), which is processed to form a short stem-loop molecule, termed a precursor microRNA (pre-miRNA), followed by endonucleolytic cleavage to form the mature microRNA. Mature microRNAs are approximately 19-24 nucleotides in length and are partially complementary to the 3′UTR (or other regions such as introns, exons or 5′UTR) of one or more target messenger RNAs (mRNAs).

A nomenclature scheme has been well established for microRNAs (Griffiths-Jones et al., Nucleic Acids Res. 34:D140-D144, 2006; Ambros et al., RNA 9:277-279, 2003; Griffiths-Jones, Nucleic Acids Res. 32:D109-D111, 2004). For example, a microRNA name includes a three or four letter species prefix, such as “hsa” for Homo sapiens, and a numeric suffix, such as “150,” resulting in a complete name of “hsa-miR-150.” Mature miRNA sequences expressed from more than one hairpin precursor molecule are distinguished by “-1” and “-2” (such as miR-6-1 and miR-6-2). Related hairpin loci expressing related mature microRNA sequences have lettered suffixes (such as miR-181a and miR-181b). In some cases, mature miRNAs from both the 5′ and 3′ arms of the hairpin precursor are identified, which are designated “3p” or “5p” (such as miR-768-3p and miR-768-5p).

MicroRNA gene product sequences are well described throughout the scientific and patent literature and are available online through miRBase (world wide web at mirbase.org), provided by the University of Manchester (previously provided by the Sanger Institute). The miRBase registry provides the nucleotide sequences of all published animal, plant and viral microRNAs (Griffiths-Jones et al., Nucleic Acids Res. 36:D154-D158, 2008). Provided by miRBase are the sequences of precursor microRNAs (stem-loop miRNAs), mature miRNAs and minor microRNA species (miR*). Precursor miRNAs predominantly express one species of miRNA, referred to as the mature miRNA. However, minor miRNA sequences have also been detected and are referred to as miR**, which are sometimes identified by their “3p” or “5p” annotation, as described above.

In the context of the present disclosure, administering a “miR” to a subject or contacting a cell with a “miR” encompasses administration or contacting with a pri-miRNA, pre-miRNA or mature miRNA, or a nucleic acid molecule encoding a pri-miRNA, pre-miRNA or mature miRNA.

Operably linked: A first nucleic acid sequence is operably linked with a second nucleic acid sequence when the first nucleic acid sequence is placed in a functional relationship with the second nucleic acid sequence. For instance, a promoter is operably linked to a coding sequence if the promoter affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary to join two protein-coding regions, in the same reading frame.

Pharmaceutically acceptable vehicles: The pharmaceutically acceptable carriers (vehicles) useful in this disclosure are conventional. Remington's Pharmaceutical Sciences, by E. W. Martin, Mack Publishing Co., Easton, Pa., 15th Edition (1975), describes compositions and formulations suitable for pharmaceutical delivery of one or more therapeutic compounds, molecules or agents (such as a miR or vector encoding a miR).

In general, the nature of the carrier will depend on the particular mode of administration being employed. For instance, parenteral formulations usually comprise injectable fluids that include pharmaceutically and physiologically acceptable fluids such as water, physiological saline, balanced salt solutions, aqueous dextrose, glycerol or the like as a vehicle. For solid compositions (for example, powder, pill, tablet, or capsule forms), conventional non-toxic solid carriers can include, for example, pharmaceutical grades of mannitol, lactose, starch, or magnesium stearate. In addition to biologically-neutral carriers, pharmaceutical compositions to be administered can contain minor amounts of non-toxic auxiliary substances, such as wetting or emulsifying agents, preservatives, and pH buffering agents and the like, for example sodium acetate or sorbitan monolaurate.

Preventing, treating or ameliorating a disease: “Preventing” a disease refers to inhibiting the full development of a disease. “Treating” refers to a therapeutic intervention that ameliorates a sign or symptom of a disease or pathological condition after it has begun to develop. “Ameliorating” refers to the reduction in the number or severity of signs or symptoms of a disease.

Promoter: A promoter is an array of nucleic acid control sequences that directs transcription of a nucleic acid. A promoter includes necessary nucleic acid sequences near the start site of transcription, such as in the case of a polymerase II type promoter (a TATA element). A promoter also optionally includes distal enhancer or repressor elements which can be located as much as several thousand base pairs from the start site of transcription. Both constitutive and inducible promoters are included (see e.g., Bitter et al., Methods in Enzymology 153:516-544, 1987).

Sequence identity/similarity: The identity/similarity between two or more nucleic acid sequences, or two or more amino acid sequences, is expressed in terms of the identity or similarity between the sequences. Sequence identity can be measured in terms of percentage identity; the higher the percentage, the more identical the sequences are. Sequence similarity can be measured in terms of percentage similarity (which takes into account conservative amino acid substitutions); the higher the percentage, the more similar the sequences are. Homologs or orthologs of nucleic acid or amino acid sequences possess a relatively high degree of sequence identity/similarity when aligned using standard methods. This homology is more significant when the orthologous proteins or cDNAs are derived from species which are more closely related (such as human and mouse sequences), compared to species more distantly related (such as human and C. elegans sequences).

Methods of alignment of sequences for comparison are well known in the art. Various programs and alignment algorithms are described in: Smith & Waterman, Adv. Appl. Math. 2:482, 1981; Needleman & Wunsch, J. Mol. Biol. 48:443, 1970; Pearson & Lipman, Proc. Natl. Acad. Sci. USA 85:2444, 1988; Higgins & Sharp, Gene, 73:237-44, 1988; Higgins & Sharp, CABIOS 5:151-3, 1989; Corpet et al., Nuc. Acids Res. 16:10881-90, 1988; Huang et al. Computer Appls. in the Biosciences 8, 155-65, 1992; and Pearson et al., Meth. Mol. Bio. 24:307-31, 1994. Altschul et al., J. Mol. Biol. 215:403-10, 1990, presents a detailed consideration of sequence alignment methods and homology calculations.

The NCBI Basic Local Alignment Search Tool (BLAST) (Altschul et al., J. Mol. Biol. 215:403-10, 1990) is available from several sources, including the National Center for Biological Information (NCBI) and on the internet, for use in connection with the sequence analysis programs blastp, blastn, blastx, tblastn and tblastx. Additional information can be found at the NCBI web site.

Recombinant: A recombinant nucleic acid molecule is one that has a sequence that is not naturally occurring or has a sequence that is made by an artificial combination of two otherwise separated segments of sequence. This artificial combination can be accomplished by chemical synthesis or by the artificial manipulation of isolated segments of nucleic acid molecules, such as by genetic engineering techniques.

Subject: Living multi-cellular vertebrate organisms, a category that includes human and non-human mammals.

Therapeutically effective amount: A quantity of a specified composition, pharmaceutical or therapeutic agent (such as a miR or nucleic acid molecule encoding a miR) sufficient to achieve a desired effect in a subject, or in a cell, being treated with the agent. The effective amount of the agent will be dependent on several factors, including, but not limited to the subject being treated, the disease or condition being treated, and the manner of administration of the therapeutic composition. In some embodiments of the present disclosure, the therapeutically effective amount (or effective amount) of a miR is the amount required to increase viral resistance or induce autophagy.

Transduce, transform or transfect: To introduce a nucleic acid molecule into a cell, such as a miR or a vector encoding a miR. These terms encompass all techniques by which a nucleic acid molecule can be introduced into a cell, including but not limited to, transduction with viral vectors, transfection with plasmid vectors, liposomal-mediated transfection and introduction of naked DNA by electroporation and particle gun acceleration. A transfected or transformed cell is a cell into which has been introduced a nucleic acid molecule by molecular biology techniques. In some examples, the nucleic acid molecule becomes stably replicated by the cell, for example by incorporation of the nucleic acid molecule into the cellular genome, or by episomal replication. In other examples, the nucleic acid molecule is transiently expressed in the cell.

Vector: A vector is a nucleic acid molecule allowing insertion of foreign nucleic acid without disrupting the ability of the vector to replicate and/or integrate in a host cell. A vector can include nucleic acid sequences that permit it to replicate in a host cell, such as an origin of replication. A vector can also include one or more selectable marker genes and other genetic elements. An expression vector is a vector that contains the necessary regulatory sequences to allow transcription and translation of inserted gene or genes. In some embodiments herein, the vector is a plasmid vector. In other embodiments, the vector is a viral vector.

Unless otherwise explained, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. The singular terms “a,”“an,” and “the” include plural referents unless context clearly indicates otherwise. Similarly, the word “or” is intended to include “and” unless the context clearly indicates otherwise. Hence “comprising A or B” means including A, or B, or A and B. It is further to be understood that all base sizes or amino acid sizes, and all molecular weight or molecular mass values, given for nucleic acids or polypeptides are approximate, and are provided for description. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, suitable methods and materials are described below. All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including explanations of terms, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting.

III. Introduction

Described herein is the finding that cultured primary human trophoblasts (PHT) are highly resistant to infection by diverse and unrelated viruses. The studies disclosed herein also determined that conditioned PHT culture medium confers resistance to viral infections in permissive non-placental cells, indicating that trophoblast-associated viral resistance is transmissible, and is transferred to recipient cells. It was found that a unique cluster of primate-specific microRNAs (miRNAs), which are highly expressed in human trophoblasts from the human chromosome 19 miRNA cluster (C19MC) (Noguer-Dance et al., Hum Mol Genet 19, 3566-3582, 2010), are packaged within PHT-derived exosomes and confer this viral resistance to recipient cells. It is further shown herein that PHT cells exhibit high rates of resting autophagy, a process involved in the maintenance of cellular homeostasis and an effective cellular countermeasure to suppress viral infections. PHT-derived exosomes and several C19MC miRNAs robustly induce autophagy in non-placental recipient cells, which is required for their resistance to viral infection. Unlike the other viruses that were tested, CMV infection is greatly enhanced by C19MC miRNAs. These findings illuminate a previously unknown pathway employed by human trophoblasts to suppress viral infections and confer viral resistance to non-placental recipient cells, suggesting a novel mechanism for shielding the placenta and maternal-derived recipient cells against viral infections during pregnancy.

IV. Overview of Several Embodiments

Provided herein is a method of inhibiting or treating a microbial infection in a subject. In some embodiments, the method includes selecting a subject with a microbial infection, or at risk for contracting a microbial infection, and administering to the subject a therapeutically effective amount of one or more microRNAs (miRs) encoded by the chromosome 19 miRNA cluster (C19MC). In some embodiments, the method includes direct administration of the one or more miRs encoded by the C19MC. In other embodiments, administering the one or more miRs encoded by the C19MC comprises administering a nucleic acid molecule encoding the C19MC or a biologically active portion thereof. In some examples, the nucleic acid molecule comprises a vector, such as a plasmid vector or a viral vector.

In some embodiments, the microbial infection is a viral infection. The viral infection can be an infection caused by any type of virus. In some examples, the virus is an RNA virus. RNA viruses include, for example, coxsackieviruses (e.g. coxsackievirus A and coxsackievirus B), poliovirus, vesicular stomatitis virus, human immunodeficiency virus, hepatitis C virus, rubella virus and morbilliviruses (such as measles virus). In particular examples, the RNA virus is a coxsackievirus, poliovirus, vesicular stomatitis virus, human immunodeficiency virus or hepatitis C virus. In other examples, the virus is a DNA virus. DNA viruses include, for example, vaccinia virus, herpes simplex viruses (HSV-1 and -2), Epstein-Barr virus, hepatitis B virus, parvovirus and varicella zoster. In particular examples, the DNA virus is a vaccinia virus or a herpes simplex virus. In some embodiments, the virus is not cytomegalovirus (CMV).

In some embodiments, the microbial infection is a bacterial infection. In some examples, the bacteria is Staphylococcus aureus, Group A Streptococcus, Listeria monocytogenes, Bacillus anthracis, Burkholderia pseudomallei, Helicobacter pylori, Salmonella enterica or Vibrio cholerae.

In some embodiments, the microbial infection is a parasitic infection. In some examples, the parasite is the protozoan parasite Toxoplasma gondii.

In some embodiments, inhibiting the microbial infection comprises preventing the microbial infection.

In some embodiments, the method comprises inhibiting or preventing intrauterine transmission of the microbial infection.

In some embodiments of the disclosed methods, administration is extrauterine. In other embodiments, administration is intrauterine.

In some examples, the one or more miRs, or nucleic acid molecule encoding the C19MC or biologically active portion thereof, is administered prophylactically to prevent infection. In other examples, the one or more miRs, or nucleic acid molecule encoding the C19MC or biologically active portion thereof, is administered to treat an existing infection.

The one or more miRs administered to the subject can be any single miR or any combination of two or more miRs that are encoded by the C19MC. Similarly, if the subject is administered a nucleic acid molecule comprising the C19MC or biologically active portion thereof, the subject can be administered the entire C19MC or a portion that encodes a single or multiple miRs. The miR genes included in the C19MC are shown in FIG. 8 and listed in Table 1. Table 1 also lists 58 unique mature miR sequences encoded by the miR genes.

In some embodiments, the subject is administered a single miR. In other embodiments, the subject is administered at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, or at least 55 miRs. In another embodiment, the subject is administered all miRs encoded by the cluster. In particular examples, the miRs are mature miRs.

In other embodiments, the subject is administered a nucleic acid molecule comprising all miR genes of the C19MC (see Table 1). In another embodiment, the subject is administered a nucleic acid molecule encoding a single miR encoded by the C19MC. In yet other embodiments, the subject is administered a nucleic acid molecule encoding at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50, or at least 55 miRs of the C19MC. In particular examples, the miRs are mature miRs.

In some embodiments, the subject is administered the entire C19MC or a nucleic acid molecule encoding the entire C19MC.

In some embodiments, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-517-3p, miR-516b-5p or miR-512-3p. In particular examples, the biologically active portion of the C19MC, or the one or more miRs comprises miR-517-3p, miR-516b-5p, and miR-512-3p. In other particular examples, the biologically active portion of the C19MC, or the one or more miRs comprises miR-517-3p, miR-1323, miR-516b-5p, miR-525-5p, miR-512-3p and miR-515-3p.

In some examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-512-3p, miR-516b, miR-517-3p, or any combination of two or more thereof. In particular examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-512-3p, miR-516b, miR- and 517-3p. In further examples, a single miR selected from miR-512-3p, miR-516b, miR- and 517-3p is administered to the subject.

In other examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-512-3p, miR-515-3p, miR-516b, miR-517-3p, miR-525-5p and miR-1323. In yet other examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-1323, miR-512-3p, miR-512-5p, miR-515-3p, miR-515-5p, miR-516b, miR-517-3p, miR-517c, miR-518a-5p, miR-518b, miR-518e, miR-519c-3p, miR-519d, miR-520c-3p, miR-520h and miR-525-5p.

In some examples, the nucleotide sequence of the one or more miRs is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to one or more of SEQ ID NOs: 1-59. In non-limiting examples, the nucleotide sequence of the one or more miRs comprises or consists of one or more of SEQ ID NOs: 1-59.

Further provided herein is a method of inducing autophagy in a cell. In some embodiments, the method includes contacting the cell with an effective amount of one or more miRs encoded by the C19MC. In some embodiments, the method includes directly contacting the cell with the one or more miR molecules. In other embodiments, contacting the cell with the one or more miRs encoded by the C19MC comprises contacting the cell with a nucleic acid molecule encoding the C19MC or a biologically active portion thereof. In some examples, the nucleic acid molecule comprises a vector, such as a plasmid vector a viral vector.

In some embodiments, the method is an in vitro method. In some examples of the in vitro method, the cell is a primary cell. In other examples, the cell is an immortalized cell.

In other embodiments, the method is an in vivo method and contacting the cell comprises administering to a subject an effective amount of one or more miRs encoded by the C19MC, or a nucleic acid molecule encoding the C19MC or a biologically active portion thereof. In some embodiments, the subject suffers from a disease associated with a deficiency in autophagy and/or a disease that can be ameliorated by stimulating autophagy. In particular examples, the disease is inflammatory bowel disease, Crohn's disease, alcoholic liver disease, Parkinson's disease, Alzheimer's disease, heart disease, diabetes or obesity.

The one or more miRs contacted with the cell (in vitro or in vivo) can be any single miR or any combination of two or more miRs that are encoded by the C19MC. Similarly, if the cell is contacted with a nucleic acid molecule encoding the C19MC or biologically active portion thereof, the cell can be contacted with the entire C19MC or a portion that encodes a single or multiple miRs. In some embodiments, the cell is contacted with a single miR. In other embodiments, the cell is contacted with at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50 or at least 55 miRs. In another embodiment, the cell is contacted with all miRs encoded by the C19MC. In particular examples, the miRs are mature miRs.

In other embodiments, the cell is contacted with a nucleic acid molecule comprising all miR genes of the C19MC (see Table 1). In another embodiment, the cell is contacted with a nucleic acid molecule encoding a single miR of the C19MC. In yet other embodiments, the cell is contacted with a nucleic acid molecule encoding at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 11, at least 12, at least 13, at least 14, at least 15, at least 16, at least 17, at least 20, at least 25, at least 30, at least 35, at least 40, at least 45, at least 50 or at least 55 miRs of the C19MC. In particular examples, the miRs are mature miRs.

In some embodiments of the methods of inducing autophagy, the subject is administered the entire C19MC or a nucleic acid molecule encoding the entire C19MC.

In some embodiments of the methods of inducing autophagy, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-517-3p, miR-516b-5p or miR-512-3p. In particular examples, the biologically active portion of the C19MC, or the one or more miRs comprises miR-517-3p, miR-516b-5p, and miR-512-3p. In other particular examples, the biologically active portion of the C19MC, or the one or more miRs comprises miR-517-3p, miR-1323, miR-516b-5p, miR-525-5p, miR-512-3p and miR-515-3p.

In some examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-512-3p, miR-516b, miR-517-3p, or any combination of two or more thereof. In particular examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-512-3p, miR-516b, and miR-517-3p. In further examples, a single miR selected from miR-512-3p, miR-516b, and miR-517-3p is contacted with the cell.

In other examples of the methods of inducing autophagy, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-512-3p, miR-515-3p, miR-516b, miR-517-3p, miR-525-5p and miR-1323. In yet other examples, the biologically active portion of the C19MC, or the one or more miRs, comprises miR-1323, miR-512-3p, miR-512-5p, miR-515-3p, miR-515-5p, miR-516b, miR-517-3p, miR-517c, miR-518a-5p, miR-518b, miR-518e, miR-519c-3p, miR-519d, miR-520c-3p, miR-520h and miR-525-5p.

In some examples of the methods of inducing autophagy, the nucleotide sequence of the one or more miRs is at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to one or more of SEQ ID NOs: 1-59. In non-limiting examples, the nucleotide sequence of the one or more miRs comprises or consists of one or more of SEQ ID NOs: 1-59.

In some embodiments of the methods disclosed herein, the one or more miRs, or the nucleic acid molecule encoding the C19MC or biologically active portion thereof, is administered to the subject or contacted with the cell using a liposomal formulation, a cationic lipid or a polypeptide carrier.

In some embodiments of the disclosed methods, the nucleic acid molecule encoding the C19MC or biologically active portion thereof comprises a vector. In some examples, the vector is a plasmid vector. In other examples, the vector is a viral vector. Viral vectors can be of, for example, adenovirus, adeno-associated virus, retrovirus, herpes virus or vaccinia virus origin. Viral vectors can include modified versions of the viruses, such as replication deficient viruses. Suitable vectors, such as gene therapy vectors, are well known in the art. In some examples, the miR is expressed from recombinant circular or linear DNA plasmids using any suitable promoter. Suitable promoters for expressing RNA from a plasmid include, for example, the U6 or H1 RNA pol III promoter sequences, or a cytomegalovirus promoter. Selection of other suitable promoters is within the skill in the art. The recombinant plasmids of the present disclosure can also comprise inducible or regulatable promoters for expression of the miR.

In some embodiments, such as when the miR is administered as a naked nucleic acid molecule, the miR includes modifications, such as nucleotide modifications to increase nuclease resistance, or other modifications to enhance delivery and/or activity of the miR.

The disclosed methods comprise administering a therapeutically effective amount, or contacting a cell with an effective amount, of at least one miR encoded by the C19MC. In some embodiments, the miR is a variant or biologically-active fragment of the miR encoded by the C19MC. Thus, the miR that is administered to a subject or contacted with a cell can be identical to an endogenous (wild-type) miR (including a pri-miRNA, pre-miRNA or mature miRNA) that is encoded by the C19MC, or it can be a variant or biologically-active fragment thereof. As defined herein, a “variant” of a miR refers to a miRNA that has less than 100% identity to a corresponding wild-type miR and possesses one or more biological activities of the corresponding miR. Examples of such biological activities include, but are not limited to, inhibition of expression of a target RNA molecule (e.g., inhibiting translation of a target RNA molecule, modulating the stability of a target RNA molecule, or inhibiting processing of a target RNA molecule), promoting viral resistance or inducing autophagy. The miR variants include species variants and variants that are the consequence of one or more mutations (e.g., a substitution, a deletion, an insertion) in a miR gene. In certain embodiments, the variant is at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at about 99% identical to a corresponding wild-type miR (such as one of the miRs listed in Table 1 or set forth herein as SEQ ID NOs: 1-59).

As used herein, a “biologically-active fragment” of a miR refers to an RNA fragment of a miR that possesses one or more biological activities of a corresponding wild-type miR. As described above, examples of such biological activities include, but are not limited to, inhibition of expression of a target RNA molecule, repressing protein translation, promoting viral resistance or inducing autophagy. In certain embodiments, the biologically-active fragment is at least about 9, at least about 11, at least about 13, at least about 15, at least about 17 or at least about 19 nucleotides in length.

V. Placental miRNAs from Chromosome 19 miRNA Cluster (C19MC)

Akin to other tissues, trophoblast differentiated functions are controlled by transcription factors, epigenetic modifiers and post-transcriptional influences that ultimately shape mRNA and protein expression. These pathways include small modulatory RNAs that interact with target gene 3′UTRs (or other regions of the genes) and promote RNA degradation and translational repression (Ghildiyal et al., Nat Rev Genet 10:94-108, 2009; Huntzinger and Izaurralde, Nat Rev Genet 12:99-110, 2011; Krol et al., Nat Rev Genet 11:597-610, 2010; Friedman et al., Genome Res 19:92-105, 2009; Thomas and Lieberman, Nat Struct Mol Biol 17:1169-1174, 2010; Vickers et al., Nat Cell Biol 13:423-433, 2011; Herranz and Cohen, Genes Dev 24:1339-1344, 2010; Bartel, Cell 136:215-233, 2009; Carthew and Sontheimer, Cell 136:642-655, 2009). Among all small RNAs, previous studies have shown that human trophoblasts produce primarily miRNAs throughout pregnancy, as well as other small RNAs (piRNAs, snRNAs, and snoRNAs) (Mouillet et al., Placenta 31:781-784, 2010; Luo et al., Biol Reprod 81:717-729, 2009; Mouillet et al., Birth Defects Res A Clin Mol Teratol 91:737-743, 2011; Barad et al., Genome Res 14:2486-2494, 2004; Pineles et al., Am J Obstet Gynecol 196:261, 2007). Many of these miRNAs are stably released into the maternal circulation, suggesting a miRNA-based mechanism for fetal-maternal communication (Mouillet et al., Placenta 31:781-784, 2010; Chim et al., Clin Chem 54:482-190, 2008).

Approximately 30-40% of placental miRNA species are expressed from defined miRNA clusters (Luo et al., Biol Reprod 81:717-729, 2009; Liang et al., Genomics 8:166, 2007; Bortolin-Cavaille et al., Nucleic Acids Res 37:3464-3473, 2009). Unique among these clusters is a primate-specific, large miRNA cluster (˜100 kb, 46 highly-related miRNA genes), expressed from chromosomal region 19q13.41 (Bentwich et al., Nat Genet 37:766-770, 2005). MiRNA members of this chromosome 19 miRNA cluster (C19MC) are expressed throughout human pregnancy, and nearly exclusively in the placenta (Luo et al., Biol Reprod 81:717-729, 2009; Liang et al., Genomics 8:166, 2007; Bortolin-Cavaille et al., Nucleic Acids Res 37:3464-3473, 2009; Chiu et al., Clin Chem 52:313-316, 2006). Although the C19MC genomic sequence contains many primate-specific Alu repeats, which may mediate gene rearrangement (Zhang et al., Mol Biol Evol 25:1493-1502, 2008; Lehnert et al., PLoS One 4:e4456, 2009), secondary structure conservation and low SNP frequency of C19MC suggests that this cluster is evolutionarily stable. Selected placenta-specific miRNA species are found in the maternal blood throughout pregnancy. Their levels are not uniform, and rapidly decline in the first 24 h postpartum (Ng et al., Proc Natl Acad Sci USA 100:4748-4753, 2003; Gilad et al., PLoS One 3:e3148, 2008).

A schematic of the genomic organization of C19MC is shown in FIG. 8. A list of the specific miRNA genes included in this cluster is provided in the table below. The names and sequences of the mature forms of each miRNA gene are also shown. Many miR genes encode more than one mature product (the “5p” and “3p” products from the 5′ and 3′ arms, respectively, of the hairpin precursor). In some instances, two different miR genes encode a mature product with the same sequence (for example, see hsa-miR-512-1 and hsa-miR-512-2). All sequences shown in Table 1 were obtained from miRBase (world wide web at mirbase.org) on Feb. 27, 2012. The present disclosure contemplates the use of any microRNA product (a pri-mRNA, pre-miRNA or mature RNA) encoded by any of the genes listed in Table 1.


TABLE 1
miRNAs of the chromosome 19 miRNA cluster
SEQ ID
Gene
Mature miRNA
Sequence of Mature miRNA
NO:
hsa-mir-512-1
hsa-miR-512-5p
CACUCAGCCUUGAGGGCACUUUC
 1
hsa-miR-512-3p
AAGUGCUGUCAUAGCUGAGGUC
 2
hsa-mir-512-2
hsa-miR-512-5p
CACUCAGCCUUGAGGGCACUUUC
 1
hsa-miR-512-3p
AAGUGCUGUCAUAGCUGAGGUC
 2
hsa-mir-1323
hsa-miR-1323
UCAAAACUGAGGGGCAUUUUCU
 3
hsa-mir-498
hsa-miR-498
UUUCAAGCCAGGGGGCGUUUUUC
 4
hsa-mir-520e
hsa-miR-520e
AAAGUGCUUCCUUUUUGAGGG
 5
hsa-mir-515-1
hsa-miR-515-5p
UUCUCCAAAAGAAAGCACUUUCUG
 6
hsa-miR-515-3p
GAGUGCCUUCUUUUGGAGCGUU
 7
hsa-mir-519e
hsa-miR-519e-5p
UUCUCCAAAAGGGAGCACUUUC
 8
hsa-miR-519e-3p
AAGUGCCUCCUUUUAGAGUGUU
 9
hsa-mir-520f
hsa-miR-520f
AAGUGCUUCCUUUUAGAGGGUU
10
hsa-mir-515-2
hsa-miR-515-5p
UUCUCCAAAAGAAAGCACUUUCUG
 6
hsa-miR-515-3p
GAGUGCCUUCUUUUGGAGCGUU
 7
hsa-mir-519c
hsa-miR-519c-5p
CUCUAGAGGGAAGCGCUUUCUG
11
hsa-miR-519c-3p
AAAGUGCAUCUUUUUAGAGGAU
12
hsa-mir-1283-1
hsa-miR-1283
UCUACAAAGGAAAGCGCUUUCU
13
hsa-mir-520a
hsa-miR-520a-5p
CUCCAGAGGGAAGUACUUUCU
14
hsa-miR-520a-3p
AAAGUGCUUCCCUUUGGACUGU
15
hsa-mir-526b
hsa-miR-526b-5p
CUCUUGAGGGAAGCACUUUCUGU
16
hsa-miR-526b-3p
GAAAGUGCUUCCUUUUAGAGGC
17
hsa-mir-519b
hsa-miR-519b-5p
CUCUAGAGGGAAGCGCUUUCUG
18
hsa-miR-519b-3p
AAAGUGCAUCCUUUUAGAGGUU
19
hsa-mir-525
hsa-miR-525-5p
CUCCAGAGGGAUGCACUUUCU
20
hsa-miR-525-3p
GAAGGCGCUUCCCUUUAGAGCG
21
hsa-mir-523
hsa-miR-523-5p
CUCUAGAGGGAAGCGCUUUCUG
22
hsa-miR-523-3p
GAACGCGCUUCCCUAUAGAGGGU
23
hsa-mir-518f
hsa-miR-518f-5p
CUCUAGAGGGAAGCACUUUCUC
24
hsa-miR-518f-3p
GAAAGCGCUUCUCUUUAGAGG
25
hsa-mir-520b
hsa-miR-520b
AAAGUGCUUCCUUUUAGAGGG
26
hsa-mir-518b
hsa-miR-518b
CAAAGCGCUCCCCUUUAGAGGU
27
hsa-mir-526a-1
hsa-miR-526a
CUCUAGAGGGAAGCACUUUCUG
28
hsa-mir-520c
hsa-miR-520c-5p
CUCUAGAGGGAAGCACUUUCUG
29
hsa-miR-520c-3p
AAAGUGCUUCCUUUUAGAGGGU
30
hsa-mir-518c
hsa-miR-518c-5p
UCUCUGGAGGGAAGCACUUUCUG
31
hsa-miR-518c-3p
CAAAGCGCUUCUCUUUAGAGUGU
32
hsa-mir-524
hsa-miR-524-5p
CUACAAAGGGAAGCACUUUCUC
33
hsa-miR-524-3p
GAAGGCGCUUCCCUUUGGAGU
34
hsa-mir-517-3p
hsa-miR-517-5p
CCUCUAGAUGGAAGCACUGUCU
35
hsa-miR-517-3p
AUCGUGCAUCCCUUUAGAGUGU
36
hsa-mir-519d
hsa-miR-519d
CAAAGUGCCUCCCUUUAGAGUG
37
hsa-mir-521-2
hsa-miR-521
AACGCACUUCCCUUUAGAGUGU
38
hsa-mir-520d
hsa-miR-520d-5p
CUACAAAGGGAAGCCCUUUC
39
hsa-miR-520d-3p
AAAGUGCUUCUCUUUGGUGGGU
40
hsa-mir-520g
hsa-miR-520g
ACAAAGUGCUUCCCUUUAGAGUGU
42
hsa-mir-516b-2
hsa-miR-516b-5p
AUCUGGAGGUAAGAAGCACUUU
43
hsa-miR-516b-3p
UGCUUCCUUUCAGAGGGU
44
hsa-mir-526a-2
hsa-miR-526a
CUCUAGAGGGAAGCACUUUCUG
28
hsa-mir-518e
hsa-miR-518e-5p
CUCUAGAGGGAAGCGCUUUCUG
45
hsa-miR-518e-3p
AAAGCGCUUCCCUUCAGAGUG
46
hsa-mir-518a-1
hsa-miR-518a-5p
CUGCAAAGGGAAGCCCUUUC
47
hsa-miR-518a-3p
GAAAGCGCUUCCCUUUGCUGGA
48
hsa-mir-518d
hsa-miR-518d-5p
CUCUAGAGGGAAGCACUUUCUG
49
hsa-miR-518d-3p
CAAAGCGCUUCCCUUUGGAGC
50
hsa-mir-516b-1
hsa-miR-516b-5p
AUCUGGAGGUAAGAAGCACUUU
43
hsa-miR-516b-3p
UGCUUCCUUUCAGAGGGU
44
hsa-mir-518a-2
hsa-miR-518a-5p
CUGCAAAGGGAAGCCCUUUC
47
hsa-miR-518a-3p
GAAAGCGCUUCCCUUUGCUGGA
48
hsa-mir-517c
hsa-miR-517-5p
CCUCUAGAUGGAAGCACUGUCU
35
hsa-miR-517c-3p
AUCGUGCAUCCUUUUAGAGUGU
51
hsa-mir-520h
hsa-miR-520h
ACAAAGUGCUUCCCUUUAGAGU
52
hsa-mir-521-1
hsa-miR-521
AACGCACUUCCCUUUAGAGUGU
38
hsa-mir-522
hsa-miR-522-5p
CUCUAGAGGGAAGCGCUUUCUG
53
hsa-miR-522-3p
AAAAUGGUUCCCUUUAGAGUGU
54
hsa-mir-519a-1
hsa-miR-519a-5p
CUCUAGAGGGAAGCGCUUUCUG
55
hsa-miR-519a-3p
AAAGUGCAUCCUUUUAGAGUGU
56
hsa-mir-527
hsa-miR-527
CUGCAAAGGGAAGCCCUUUC
57
hsa-mir-516a-1
hsa-miR-516a-5p
UUCUCGAGGAAAGAAGCACUUUC
58
hsa-miR-516a-3p
UGCUUCCUUUCAGAGGGU
59
hsa-mir-1283-2
hsa-miR-1283
UCUACAAAGGAAAGCGCUUUCU
13
hsa-mir-516a-2
hsa-miR-516a-5p
UUCUCGAGGAAAGAAGCACUUUC
58
hsa-miR-516a-3p
UGCUUCCUUUCAGAGGGU
59
hsa-mir-519a-2
hsa-miR-519a-3p
AAAGUGCAUCCUUUUAGAGUGU
56

Placental Exosomes

A significant fraction of plasma miRNA is packaged in exosomes. These nanoparticles function as “cargo vehicles” (Valadi et al., Nat Cell Biol 9:654-659, 2007; Simpson et al., Proteomics 8:4083-4099, 2008; Skog et al., Nat Cell Biol 10:1470-1476, 2008) to transfer nucleic acids, proteins, lipids and other biomolecules to proximal and distant tissues (Thery et al., Nat Rev Immunol 9:581-593, 2009; Raposo et al., J Exp Med 183:1161-1172, 1996; Alvarez-Erviti et al., Nat Biotechnol 29:341-345, 2011). Exosomes belong to a large family of intracellular and extracellular microvesicular bodies, ranging in size between >100 nm (apoptotic blebs and microparticles) to approximately 30-120 nm (exosomes). Exosomes can fuse intracellularly with lysosomes to destroy content or fuse with plasma membranes to release exosomal content via exocytosis or ectocytosis (Thery et al., Nat Rev Immunol 2:569-579, 2002; Pan and Johnstone, Cell 33:967-978, 1983; Keller et al., Immunol Lett 107:102-108, 2006). Exosomes are defined by size, cup-shaped form, sucrose gradient buoyancy (1.13-1.19 g/mL), and a detergent-resistant, lipid raft-rich membrane bilayer of proteins, cholesterol, and sphingolipids. Some of the membrane proteins are common, such as cytoskeletal proteins, chaperones, and tetraspanins, including CD9, CD63, CD81, and others specific to the exosome's cell of origin (Simpson et al., Proteomics 8:4083-4099, 2008; Thery et al., Nat Rev Immunol 2:569-579, 2002). Surface proteins may determine exosome specificity to target cells, which they enter via endocytosis-based internalization (Morelli et al., Blood 104:3257-3266, 2004), cell membrane fusion (Denzer et al., J Immunol 165:1259-1265, 2000), or receptor-ligand interactions (Admyre et al., Eur J Immunol 36:1772-1781, 2006).

Thus, exosomes act as a form of communication among different cell types, with potentially striking consequences to recipient cells (e.g., induction of apoptosis, mediated by the FAS ligand binding to FAS receptors). Relevant to the present disclosure, exosomes were recently found to be enriched for miRNAs (Valadi et al., Nat Cell Biol 9:654-659, 2007; Eldh et al., PLoS One 5:e15353, 2010), akin to virally-mediated intercellular transfer of genetic material, with potential beneficial or harmful consequences (Eldh et al., PLoS One 5:e15353, 2010). The human placenta is known to express microvesicular bodies of diverse sizes, mainly shed as syncytiotrophoblast microparticles, implicated in preeclampsia-related placental apoptosis. Production of exosomes has been studied in first trimester trophoblasts, where exosomes likely contribute to the establishment of maternal immune tolerance, possibly via impaired T-cell signaling, down-regulation of NK cell receptor NKG2D, and enhanced apoptotic pathways through FasL, TRAIL, and PD-L (Luo et al., Biol Reprod 81:717-729, 2009; Taylor et al., J Immunol 176:1534-1542, 2006; Hedlund et al., J Immunol 183:340-351, 2009; Mincheva-Nilsson and Baranov, Am J Reprod Immunol 63:520-533, 2010). The production and function of placental exosomes after the first trimester of human pregnancy, and the role of miRNAs packaged in these exosomes, has not been previously studied.

VI. Administration of miRNAs

A nucleic acid molecule encoding C19MC, or a biologically active portion thereof (including a single miR or multiple miRs), can be administered to a subject in need of treatment using any suitable means known in the art. Nucleic acid-based therapeutic agents can be administered to a subject by any suitable route. In some examples, the nucleic acid molecules are administered using an enteral or parenteral administration route. Suitable enteral administration routes include, for example, oral, rectal, or intranasal delivery. Suitable parenteral administration routes include, for example, intravascular administration (such as intravenous bolus injection, intravenous infusion, intra-arterial bolus injection, intra-arterial infusion and catheter instillation into the vasculature); subcutaneous injection or deposition, including subcutaneous infusion (such as by osmotic pumps); direct application to the tissue of interest, for example by a catheter or other placement device (e.g., a suppository or an implant comprising a porous, non-porous, or gelatinous material); and inhalation. In some cases, suitable administration routes are injection, infusion and direct injection into a target tissue.

In the context of the present disclosure, a miR or a nucleic acid molecule encoding C19MC, or a biologically active portion thereof, can be administered to the subject either as naked RNA or DNA in combination with a delivery reagent, or can be encoded by a recombinant plasmid or viral vector. Recombinant plasmids and viral vectors including sequences that express the C19MC or biologically active portion thereof, and techniques for delivering such plasmids and vectors to target cells, are well known in the art.

In some embodiments, liposomes are used to deliver the nucleic acid molecule to a subject. Liposomes can also increase the blood half-life of nucleic acids. Suitable liposomes for use with the present disclosure can be formed from standard vesicle-forming lipids, which generally include neutral or negatively charged phospholipids and a sterol, such as cholesterol. The selection of lipids is generally guided by consideration of several factors, such as the desired liposome size and half-life of the liposomes in the blood stream. A variety of methods are known in the art for preparing liposomes (see, for example, Szoka et al., Ann. Rev. Biophys. Bioeng. 9:467, 1980; and U.S. Pat. Nos. 4,235,871; 4,501,728; 4,837,028; and 5,019,369). In some embodiments, polymers can be used to deliver a nucleic acid molecule to a subject. Cationic lipids and polymers that can be used to deliver therapeutic nucleic acid molecules have been described (see, for example, Zhang et al., J Control Release. 123(1):1-10, 2007; Vorhies et al., Methods Mol Biol. 480:11-29, 2009; and U.S. Patent Application Publication No. 2009/0306194). Polypeptide carriers can also be used to administer nucleic acid molecules, such as miRs, to a subject (see, for example, Rahbek et al., J. Gene Med. 10:81-93, 2008).

Nucleic acid molecules can be administered in any suitable manner, preferably with pharmaceutically acceptable carriers. Pharmaceutically acceptable carriers are determined in part by the particular composition being administered, as well as by the particular method used to administer the composition. Accordingly, there is a wide variety of suitable formulations of pharmaceutical compositions of the present disclosure.

Preparations for parenteral administration include sterile aqueous or non-aqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils.

Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present such as, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like.

Formulations for topical administration may include ointments, lotions, creams, gels, drops, suppositories, sprays, liquids and powders. Conventional pharmaceutical carriers, aqueous, powder or oily bases, thickeners and the like may be necessary or desirable.

Compositions for oral administration include powders or granules, suspensions or solutions in water or non-aqueous media, capsules, sachets, or tablets. Thickeners, flavorings, diluents, emulsifiers, dispersing aids or binders may be desirable.

Administration can be accomplished by single or multiple doses. The dose required will vary from subject to subject depending on the species, age, weight and general condition of the subject, the particular nucleic acid molecule being used and its mode of administration. An appropriate dose can be determined by one of ordinary skill in the art using only routine experimentation.

The following examples are provided to illustrate certain particular features and/or embodiments. These examples should not be construed to limit the disclosure to the particular features or embodiments described.

EXAMPLES

Example 1: Materials and Methods

This example describes the materials and experimental procedures used for the studies described in Example 2.

Cells and PHT Exosome Purification

Primary human trophoblasts (PHT cells) were isolated from normal singleton term placentas using the trypsin-deoxyribonuclease-dispase/Percoll method as described by Kliman et al., with previously published modifications (Kliman et al., Endocrinology 118, 1567-1582, 1986; Nelson et al., Am J Obstet Gynecol 180, 896-902, 1999). Cells were maintained in DMEM (Sigma) containing 10% fetal bovine serum (FBS, Hyclone, Logan, Utah, USA) and antibiotics at 37° C. in a 5% carbon dioxide (CO2)-air atmosphere. Cells were maintained 72 h after plating, with cell quality monitored both morphologically (by microscopy) and by medium human chorionic gonadotropin (hCG) levels, determined by enzyme-linked immunosorbent assay (ELISA, DRG International, Mountainside, N.J.), showing a characteristic increase in medium hCG as cytotrophoblasts differentiate into syncytiotrophoblasts (Nelson et al., Am J Obstet Gynecol 180, 896-902, 1999; Chen et al., J Biol Chem 281, 2764-2772, 2006).

For isolation of PHT exosomes, PHT cells were maintained for 48 h in DMEM containing 10% FBS that was ultracentrifuged at 108,000×g for 10 h to deplete pre-existing FBS exosomes. Exosomes were isolated as described previously (Montecalvo et al., J Immunol 180, 3081-3090, 2008). Briefly, supernatants from 200 million PHT cells were combined and subsequently centrifuged at 300×g for 5 min, 1,200×g for 10 min, and 10,000×g for 30 min. Exosomes were concentrated by centrifugation at 2,500×g for 25 min using a Vivacell 100 filter (BioExpress, Kaysville, Utah, USA; F-2820-100C), then ultracentrifuged at 108,000×g for 1 h, and the exosome pellet was subsequently ultracentrifuged on top of a 30% sucrose density cushion at 108,000×g for 1 h (Lamparski et al., J Immunol Methods 270, 211-226, 2002). The exosomal phase was collected and resuspended in PBS, before ultracentrifugation at 108,000×g for 1 h. The total amount of exosomes was determined by total protein spectrophotometry. Exosome-depleted PHT supernatant was produced by subsequently centrifugation at 300×g for 5 min, 1,200×g for 10 min, 10,000×g for 30 min and 108,000×g for 1.5 h. Exosomes were reconstituted in FBS-exosome depleted complete medium at a ten-fold concentration over conditioned medium.

Human osteosarcoma U2OS, human foreskin fibroblast (HFF), and Huh7.5 cells were cultured in DMEM-H supplemented with 10% FBS and penicillin/streptomycin. Vero African green monkey kidney cells were maintained in DMEM-H supplemented with 5% FBS and penicillin/streptomycin. Caco-2 (ATCC clone) human intestinal epithelial cells were cultured in MEM supplemented with 10% FBS, non-essential amino acids, sodium pyruvate, and penicillin/streptomycin. Immortalized human first trimester extravillous trophoblast cells, provided by CH Graham, Kingston, Ontario, Canada (Graham et al., Exp Cell Res 206, 204-211, 1993) were cultured in RPMI-1640 (Cellgro, Manassas, Va., USA), supplemented with 5% bovine growth serum (HyClone) and antibiotics.

Conditioned media from PHT or other cells were harvested between 48-72 h post plating. Conditioned medium was subjected to sonication or heat-inactivation for 30 min at 65° C. Recipient cells were exposed to conditioned medium for ˜24 h prior to assay.

Viruses

Experiments were performed with vesicular stomatitis virus (VSV), green fluorescent protein (GFP)-tagged VSV, recombinant yellow fluorescent protein (YFP)-tagged vaccinia virus as described (VV) (Moser et al., PLoS Pathog 6, e1000954, 2010), coxsackievirus B3-RD isolate (CVB3-RD) as described (Coyne and Bergelson, Cell 124, 119-131, 2006), poliovirus (PV) as described (Coyne et al., EMBO J 26, 4016-4028, 2007), cytomegalovirus (hCMV Towne strain), cell culture grown hepatitis C virus (HCV) expressing firefly luciferase (HCVcc-luc), or GFP-tagged herpes simplex virus-1 (HSV1, strain KOS) as described (Desai and Person, J Virol 72, 7563-7568, 1998). VSV was expanded by growth on Vero cells and media was harvested. Viral titers were determined by plaque assays as previously described (Bozym et al., Cell Host Microbe. 11:153-166, 2012). Plaque assays were conducted on Vero (VSV and GFP-VSV) or HeLa (CVB) cells. Confluent monolayers were treated with serial dilutions of virus for one hour at 37° C. (VSV) or at room temperature (CVB). Cells were then overlayed with agarose and incubated for 48 h. Plaques were visualized by crystal violet staining and plaques enumerated. HCVcc-luc propagation was performed as described (Liu et al., J. Virol. 83:2011-2014, 2009).

Experiments assessing productive virus infection were performed as follows. PHT cells were infected with CVB, PV, VSV, VV, or HSV-1 for 14-15 h (multiplicity of infection (MOI)=5), or CMV for 24 h. Infections were performed with three individual PHT preparations in duplicate. hCMV infections were performed with two individual PHT preparations in triplicate. For 3-MA experiments assessed by RT-qPCR, PHT cells were infected with GFP-VSV for 15 h at MOI=5. For experiments analyzing immediate early viral gene expression measured by RT-qPCR, PHT cells were infected with CVB, VSV, VV, or HSV-1 for 6-7 h at MOI=1. HeLa cells were infected with CVB or PV at an MOI=5 for 8 h. HFF cells were infected with CMV for 24 h, VSV or CVB (MOI=5) for 15 h. Vero cells were infected with VSV for 6 h (MOI=5). Caco-2 cells were infected with VSV or CVB for 7 h (MOI=5). RL-95 cells were infected with CVB for 15 h (MOI=5). For immunofluorescence, U2OS cells were infected with CVB for 7 h (MOI=5), VSV (MOI=5), VV, or HSV-1 (MOI=1) for 15 h. For RT-qPCR, U2OS cells were infected with CMV, VSV, HSV-1 or VV for 5-6 h (MOI=1). Huh7.5 cells were infected with HCVcc as described previously (Liu et al., J. Virol. 83:2011-2014, 2009).

miRNA Mimics, Plasmids, and Transfections

Mimics for C19MC miRNAs (miRIDIAN) as well as a non-targeting control miRNA mimic were obtained from Thermo-Fisher (Dharmacon, Lafayette, Colo.) as described (Donker et al., Mol. Hum. Reprod. 18(8):417-424, 2012). U2OS cells or other cell lines were transfected with one or multiple miRNA mimics or miRNA mimic negative control (final concentration 6 nM for each miRNA mimic) using DharmaFECT-1 transfection reagent (Thermo Fisher Scientific) or HiPerFect™ (Qiagen) according to manufacturer's instructions. Cells were assayed 48 h post-transfection.

The total concentration of non-targeting control miRNA mimics was adjusted to that of all active miRNA mimics. For siRNA transfections, U2OS cells were reverse transfected using HiPerFect™ transfection reagent (Qiagen). For silencing of beclin-1, 40 nM per well of scrambled non-targeting siRNA (siControl) or beclin-1 siRNA (Cell Signaling, #6222S) were transfected.

Plasmid transfections were performed using X-tremeGENE 9 (Roche) according to manufacturer's protocol. The mRFP-LC3B expression construct was purchased from Addgene (plasmid 21075) and originally constructed by Tamotsu Yoshimori (Kimura et al., Autophagy 3, 452-460, 2007). For experiments with conditioned media and purified exosomes, cells were transfected, exposed to media 24 h later, and fixed 48 h post-transfection. For all other experiments, the cells were assayed 48 h post-transfection.

C19MC BAC Preparation and Transfection

The BAC RP11-1055017 containing 160,970 bp of genomic DNA from region q13-42 of chromosome 19 was obtained from the BACPAC Resource Center located at the Children's Hospital Oakland Research Institute (CHORI) in Oakland, Calif. The BAC clone harbors the entire C19MC miRNA cluster spanning around 100 kb and contains an additional 60 kb of flanking sequences. The nucleotide sequence of the genomic insert in BAC RP11-1055O17 is set forth herein as SEQ ID NO: 60. Recombineering of the BAC was performed as described (Warming et al., Nucleic Acids Res 33, e36, 2005). A GFP::zeocin cassette from the pSELECT-GFPzeo-mcs plasmid (InvivoGen, CA), was PCR amplified and cloned into the Hind III and Bam HI sites of pBluescript II SK(+) (pBS-SK). BAC specific homology arms of ˜500 bp each were PCR amplified and cloned into pBS-SK using the restriction sites flanking the GFP::zeocin cassette (5′arm: Xho I and Hind III; 3′arm: Bam HI and Xba I). The whole targeting cassette was then PCR amplified, gel purified, and electroporated into the recombinogenic SW106 bacterial strain containing the recipient BAC. In addition to the construct that was simply tagged with the GFP::zeocin cassette at the 3′end of the C19MC cluster, a BAC with a deletion of the entire C19MC coding sequence was created and used as a control in transfection experiments. By choosing a 5′ homology arm located upstream of the miRNA cluster and keeping the same 3′ homology arm, the recombination led to the actual deletion (BAC “trimming”) of the entire miRNA locus (˜100 kb). The construct contain ˜60 kb of genomic DNA flanking the GFP::zeocin cassette. Bacteria harboring the BAC with the desired alteration were selected on selective medium with chloramphenicol (12.5 μg/ml) and zeocin (25 μg/ml). The correct BAC constructs, confirmed by using restriction mapping and PCR, were transformed back into DH10B bacterial cells for propagation. BAC DNA for transfection was prepared using the PhasePrep BAC DNA kit following the recommendations from the manufacturer (Sigma).

RNA Isolation, Microarrays and miRNA RT-qPCR

For miRNA analysis, total cellular RNA was purified from cells using miRNeasy Mini Kit (Qiagen, Valencia, Calif.), according to the manufacturer's instructions. Prior to RNA isolation, non-exosomal RNA were degraded using 400 ng/μl RNase at 37° C. for 15 min (Valadi et al., Nat Cell Biol 9, 654-659, 2007). For miRNA analysis, reverse transcription and quantitative PCR (RT-qPCR) of duplicate samples was performed using the miScript PCR system (Qiagen, Valencia, Calif.), following the manufacturer's instructions. Detection of all miRNAs was performed using respective miScript primers (Qiagen, Valencia, Calif.). Dissociation curves were run on all reactions to ensure amplification of a single product. Control H2O samples were included in all RT and PCR reactions. Total RNA input was normalized using RNU6B RNA as an endogenous control. The fold increase relative to control samples was determined by the 2-ΔΔCt method (Livak and Schmittgen, Methods 25, 402-408, 2001). Microarray analysis of C19MC miRNA expression in PHT cells and in PHT exosomes was performed using Agilent's Human miRNA V3 8×15K arrays.

For analysis of C19MC miRNA targets, total cellular RNA was purified from U2OS or HTR8/Sv-Neo cells using miRNeasy Mini Kit (Qiagen), according to the manufacturer's instructions (Agilent Technologies, Santa Clara, Calif.). The quality of RNA was confirmed using 2100 Bioanalyzer (Agilent Technologies, Santa Clara, Calif.). A total of 100 ng of RNA was used for labeling, generating cyanine 3-labeled lineary amplified cRNA. Six hundred ng of cRNA was used for microarray hybridization as per Agilent protocol, performed using Agilent's SurePrint G3 Hmn GE 8×60K human arrays. Array data were extracted using a High-Resolution C scanner (Agilent) and a GE1 107 (September 9) feature extraction protocol (Agilent).

All data were from the three experimental paradigms were log 2 transformed and normalized separately using the cyclic loess normalization method (Wu et al., BMC Bioinformatics 6:309, 2005). Identical probes targeting the same mRNA transcripts were averaged by probe set intensity values. A moderated student's t test, which is based on an empirical Bayesian algorithm, as implemented in the R package “limma” (G. K. Smyth, “Linear models and empirical Bayes methods for assessing differential expression in microarray experiments,” Statistical Applications in Genetics and Molecular Biology, 3:Article 3, 2004) was applied to test, for each gene, if it was differentially expressed between the cells transfected by the empty BAC or by the C19MC BAC. The Storey's q-value method (Storey and Tibshirani, Proc Natl Acad Sci USA 100, 9440-9445, 2003) was used to calculate the adjusted p values for the p values of the moderated t test to control the false discovery rate. Up- or down-regulated genes were subsequently identified that satisfied the following conditions: (1) down (or up) regulation in the C19MC-transfected HTR8 cells with adjusted p values ≤0.05 and log 2 fold change ≤−0.5 (or ≥+0.5), (2) down (or up) regulation in the C19MC-transfected U2OS cells with adjusted p values ≤0.05 and log 2 fold change ≤−0.5 (or ≥+0.5), and (3) down (or up) regulated in the U2OS cells exposed to conditioned medium, with log 2 fold change ≤−0.5 (or ≥+0.5, note that because each group in the U2OS conditioned medium experiment had only one sample, no statistical testing was performed). Finally, the TargetScan miRNA target database (version 6) was searched to identify, among the down regulated genes, those that are predicted targets of at least one of the 4 most abundant C19MC miRNAs: miR-517-3p, miR-517b, miR-516b, and miR-512-3p. All the analyses were performed using the statistical computing program R and its packages (R Development Core Team, 2011. R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0).

For cellular mRNA analysis, total RNA was extracted using TRIreagent (MRC) or RNeasy (Qiagen) according to manufacturer's protocol. RNA samples were treated with RNAse-free DNAse (Qiagen). Total RNA was reverse transcribed using iScript cDNA synthesis kit (Bio-Rad) or RT2 First Strand kit (SABiosciences). For each sample, 0.25-1 μg RNA was used for cDNA synthesis. Real-time PCR as performed using iQ SYBR Green Supermix (Bio-Rad) in a Applied Biosystems StepOnePlus real-time PCR machine according to the manufacturer's instructions. Dissociation curves were run on all reactions to ensure amplification of a single product. Gene expression was calculated using the ΔΔCt values with normalization to human actin


(ACTGGGACGACATGGAGAAAA, SEQ ID NO: 61; 
GCCACACGCAGCTC, SEQ ID NO: 62).

Primers used were as follows: VSV (TGCAAGGAAAGCATTGAACAA, SEQ ID NO: 63; GAGGAGTCACCTGGACAATCACT, SEQ ID NO: 64), GFP (CACATGAAGCAGCACGACTTCT, SEQ ID NO: 65; AACTCCAGCAGGACCATGTGAT, SEQ ID NO: 66), hCMV Towne strain (GCGGTGGTTGCCCAACAGGA, SEQ ID NO: 67; ACGACCCGTGGTCATCTTTA, SEQ ID NO: 68), ATG4C (TAGAGGATCACGTAATTGCAGGA, SEQ ID NO: 69; GTTGTCAAAGCTGAGCCTTCTAT, SEQ ID NO: 70), UVRAG (ATGCCAGACCGTCTTGATACA, SEQ ID NO: 71; TGACCCAAGTATTTCAGCCCA, SEQ ID NO: 72), PIK3C3 (GAACAACGGTTTCGCTCTTTG, SEQ ID NO: 73; GCTTCTACATTAGGCCAGACTTT, SEQ ID NO: 74), Tk (ACCCGCTTAACAGCGTCAACA, SEQ ID NO: 75; CCAAAGAGGTGCGGGAGTTT, SEQ ID NO: 76), VV rpo35 early (GCCAATGAGGGTTCGAGTTC, SEQ ID NO: 77; AACAACATCCCGTCGTTCATC, SEQ ID NO: 78), CVB3 (ACGAATCCCAGTGTGTTTTGG, SEQ ID NO: 79; TGCTCAAAAACGGTATGGACAT, SEQ ID NO: 80), and ISG56 (CAACCAAGCAAATGTGAGGA, SEQ ID NO: 81; GGAGACTTGCCTGGTGAAAA, SEQ ID NO: 82).

Autophagy and toll-like receptor qPCR arrays (SABiosciences) were performed with 1 μg RNA per 96 well plate and subjected to RT-qPCR using SYBR/ROX RT2 qPCR 2× master mix (SABiosciences) according to manufacturer's protocol. Gene expression was defined from the threshold cycle (Ct), and relative expression levels were calculated using SABiosciences RT2 Profiler PCR array analysis automated software.

RNA library construction and miRNA sequencing was performed by Ocean Ridge Biosciences (Palm Beach Gardens, Fla.) using extracted RNA. The small RNA libraries were aligned to the NCBI-37 human reference genome using Bowtie, then intersected with the mature miRNA sequenced annotated by miRBase (v.18) using BEDtools. The miRNA counts in each library were normalized using established algorithms (Anders and Huber, Genome Biol. 11:R106, 2010). The C19MC miRNAs and the non-C19MC miRNAs in the six libraries were Laplace smoothed by adding 1 to the normalized counts, log 2 transformed, and clustered respectively by the agglomerative hierarchical clustering, using the complete linkage method. Heat maps were then generated separately for the clustered C19MC miRNAs and non-C19MC miRNAs.

To quantify the differences in miRNA expression between U2OS cells that were exposed to PHT conditioned medium vs. cells that were exposed to fresh medium, the differential expression test was applied, which assumes that the in all libraries were follow negative binomial distributions (Anders and Huber, Genome Biol. 11:R106, 2010), and a shrinkage estimator was used for the dispersion parameters of the miRNAs. The p-values of the tests were adjusted using the Benjamini and Hochberg's method (Hochberg and Benjamini, J. Roy. Statist. Soc. B. 57:289-300, 1995) to control for false discovery rate. Statistical analyses were performed using statistical computing software R and the DESeq package of R.

Pharmacological Agents

Cells were pre-treated with 3-methyladenine (3-MA; 5 mM, Sigma) for 30-60 min prior to infection, and cells were incubated with drug throughout the duration of infection. For mRFP-LC3B punctae assays, 3-MA was added for 30 min prior to conditioned or non-conditioned media exposure, and was present throughout. Rapamycin (5 μM, Calbiochem) treatment or serum-starvation with Hank's Balanced salt solution (HBSS) for 4 h was used as a positive control for autophagy.

Immunofluorescence and Confocal Microscopy

Cell monolayers were cultured in 8-well chamber slides (LabTek) at 37° C. Cells were then washed and fixed as indicated with either ice cold methanol, 3:1 methanol-acetone, or 4% paraformaldehyde in PBS and permeabilized with 0.25% Triton X-100 in PBS. Mouse anti-VSV-G and mouse anti-hCMV gB were obtained from Santa Cruz Biotechnology, and mouse anti-enterovirus VP1 (NCL-Entero) was purchased from Novacastra Laboratories. Mouse-anti clathrin heavy chain (CHC) and mouse anti-caveolin 1 (Cav1) antibodies were obtained from BD Transduction Laboratories. Rabbit anti-Dynamin II (DynII) was purchased from Abcam. Fixed monolayers were incubated with primary antibody, washed, incubated with Alexa Fluor-488 or -594-conjugated secondary antibodies (Invitrogen), washed, and then mounted with Vectashield (Vector Laboratories) containing 4′,6-diamidino-2-phenylindole (DAPI). Cholera toxin B (CTB) conjugated to Alexa Fluor 488 (8 μg/mL; Invitrogen) and transferrin conjugated to Alexa Fluor 594 (Invitrogen) uptake was performed essentially as previously described (Patel et al., J Virol 83, 11064-11077, 2009).

Images were captured with an IX81 inverted microscope equipped with a motorized stage or with an Olympus Fluoview 1000 laser scanning confocal microscope. Images of infected cells were taken using an Olympus PlanApo 10×/0.40 NA dry or Apo 20×/0.75 NA dry objective, whereas all other images were taken with an Olympus PlanApo 60×/1.42 NA oil objective.

For virus infection assays, cells were fixed and stained for markers of virus infection (CVB and PV (VP1), VSV (VSV-G), hCMV (gB)) or assessed for GFP-expression (VV-GFP, HSV-1-GFP, VV-YFP). A minimum of three independent fields per condition were counted (at least 600 cells total). Infection levels are reported as the percentage of virus positive cells among the total number of cells, determined by DAPI staining. Quantification of percent virus positive cells was performed using ImageJ (National Institutes of Health) analysis. For LC3B autophagy assays, at least twenty individual cells from a minimum of four independent fields were captured per condition. The total number of mRFP-LC3B-positive punctae were quantified per cell using ImageJ analysis with identical settings per condition. Analysis of the extent of VSV and mRFP-LC3b punctate co-localization was performed using ImageJ.

Electron Microscopy

Cells were washed, fixed with 2.5% gluteraldehyde in PBS for 1 h, then processed for electron microscopy as previously described (Gao et al., J Biol Chem 285, 1371-1383, 2010). Sections were imaged using a JEOL JEM 1011 transmission electron microscope (Peabody) using an 80 V fitted with a bottom mount AMT 2k digital camera (Advanced Microscopy Techniques). At least five to ten individual cells were captured per condition. The number of autophagosomes (including amphisomes, autophagosomes, autophagic vacuoles, and autolysosomes) were quantified per cell manually.

Immunoblots

Cells were grown in 6-well plates and lysates were prepared with RIPA buffer (50 mM Tris-HCl [pH 7.4]; 1% NP-40; 0.25% sodium deoxycholate; 150 mM NaCl; 1 mM EDTA; 1 mM phenylmethanesulfonyl fluoride; 1 mg/ml aprotinin, leupeptin, and pepstatin; 1 mM sodium orthovanadate), and insoluble material was precipitated by brief centrifugation. Protein concentration of lysates was determined by BCA protein assay (Thermo Scientific). Lysates containing equal amounts of protein were loaded onto 4-20% Tris-HCl gels (Bio-Rad) and transferred to polyvinylidene difluoride membranes. Membranes were blocked in 5% nonfat dry milk, probed with the indicated antibodies, and developed with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology), and SuperSignal West Pico or Dura, chemiluminescent substrates (Pierce Biotechnology).

For beclin-1 immunoblotting, cells were lysed on ice in a 50 mM Tris-HCl, pH 7.5 buffer that contained 150 mM NaCl and 0.5% NP-40. After centrifugation at 12,000×g at 4° C., the supernatant was subjected to 10% SDS-PAGE, transferred to PVDF membrane, and signal detected using monoclonal antibodies as indicated. Densitometry was performed using Image J.

Reporter Gene Assay

Activation of interferon β (IFNβ) or interferon-stimulated response element (ISRE) promoters was measured by reporter assay. Cells were transfected with 1 μg of DNA/well of a 24 well plate, a 30:1 ratio of IFNβ or ISRE firefly luciferase reporter plasmids to pRL-null (Renilla control) as per manufacturer's protocol. Cells were lysed in 100 μL of lysis buffer and the levels of firefly and Renilla luciferase levels quantified using the Dual-Luciferase Reporter Assay System (Promega) with a dual injector equipped Synergy 2 SL Luminescence Microplate Reader (BioTek). Levels of firefly luciferase were normalized to control Renilla luciferase levels. For poly(I:C) treatment, cells were transfected with 1 μg poly(I:C)/well using XtremeGene-9 for 16 h as per the manufacturer's protocol.

Statistical Analysis

All experiments were performed at least three times, as indicated in the figure legends. Data are presented as mean±standard deviation. Except where specified, Student's t test was used to determine statistical significance for virus infection and autophagy assays when 2 sets were compared, and one-way analysis of variance (ANOVA) with Bonferroni's correction for multiple comparisons were used to determine statistical significance for reporter gene assays. A p<0.05 was determined significant.

Virus Entry Assays

Virus entry assays in PHT cells were performed with CVB and PV as previously described (Coyne and Bergelson, Cell 124, 119-131, 2006; Coyne et al., EMBO J 26, 4016-4028, 2007). VV and HSV-1 internalization assays were performed by incubating PHT cells with virus (MOI 25) at 37° C. until fixation at various time points (30, 60, 90 min). VSV entry assays in U2OS cells exposed to either non-conditioned or conditioned PHT medium for 24 h was performed by incubating cells with virus (MOI=500) for 1 h at 37° C. until fixation in 4% PFA followed by permeabilization in 0.1% Triton X-100. VSV particles were visualized with anti-VSV-G antibody.

Modified TCID50 Virus Titering Assays

Vero or PHT cells were seeded to confluence in 96 well plates. Cells were incubated with serial dilutions of the indicated viruses for approximately 40-45 h, then stained with 0.05% crystal violet (in 10% ethanol). For experiments performed with conditioned medium, Vero cells were incubated in non-conditioned or conditioned medium 24 h prior to incubation with virus. Serial dilutions of virus were made in either non-conditioned or conditioned medium, and cells were incubated and developed with crystal violet as described above.

Neutralizing Virus Plaque Assays

VSV virus stock was diluted 1:20 in either non-conditioned or conditioned PHT medium, then incubated at 37° C. for 1 h. Plaques assays were performed on Vero cells. Plaques were visualized after 36 h by staining with crystal violet.

Example 2: Human Placental Trophoblasts Confer Viral Resistance to Recipient Cells by the Release of miRNAs and the Induction of Autophagy

This example describes the finding that PHTs are highly resistant to infection by a number of different types of viruses. This resistance is mediated by exosomes containing miRNAs encoded by the primate-specific chromosome 19 miRNA cluster (C19MC).

PHT-Derived Exosomes Protect Recipient Cells from Viral Infection

The studies described herein determined that PHT cells were resistant to infection by a panel of viruses, including coxsackievirus B3 (CVB), poliovirus (PV), vesicular stomatitis virus (VSV), vaccinia virus (VV), herpes simplex virus-1 (HSV-1), and human cytomegalovirus (CMV), when compared to non-PHT cells (FIG. 1A). This lack of viral replication was not due to inefficient viral binding and/or entry, or to defects in common endocytic pathways utilized by viruses for their entry, such as clathrin- or caveolar-mediated pathways. It was found that exposure of diverse non-PHT recipient cells for 24 h prior to infection to PHT conditioned medium (isolated from naïve PHT cells 48-72 h post-plating) decreased the replication of CVB, VSV, hepatitis C virus (HCV), and VV (FIG. 6A). The antiviral effect of conditioned PHT medium was also observed in several physiologically relevant fetal and/or maternal primary cells, including human umbilical vein endothelial cells (HUVEC), human uterine microvascular endothelial cells, human placental fibroblasts, and human foreskin fibroblasts (HFF; FIG. 1C, right panel). In contrast, conditioned medium from other cell types, such as immortalized trophoblast BeWo cells, had no effect (FIG. 6B). This effect was not the result of direct neutralization of the virus as conditioned medium had no direct effect on viral titers (FIG. 6C). Furthermore, antiviral effects were observed across multiple conditioned medium samples isolated from independent and unrelated PHT preparations (FIGS. 6D-6F). Together, these data indicate that PHT cells release specific components to the medium, which are capable of conferring viral resistance to non-placental recipient cells.

To better define the component in conditioned medium of PHT cells that is responsible for conferring viral resistance, the conditioned medium was exposed to heat inactivation or RNAse treatment; however, no effect was observed (FIG. 1D). In contrast, repeated freeze-thawing partly attenuated the effect, and sonication completely abolished the antiviral effect of PHT conditioned medium (FIG. 1D). Because exosomes, which function as “cargo nanovesicles” (Valadi et al., Nat. Cell. Biol. 9:654-659, 2007; Skog et al., Nat. Cell. Biol. 10:1470-1476, 2008), are characteristically released from trophoblasts and are sensitive to sonication (Donker et al., Mol. Hum. Reprod. 18(8):417-424, 2012; Taylor et al., J. Immunol. 176:1534-1542, 2006; Montecalvo et al., J. Immunol. 180:3081-3090, 2008; Luo et al., Biol. Reprod. 81:717-729, 2009; Pegtel et al., Proc. Natl. Acad. Sci. U.S.A. 107:6328-6333, 2010), their role in PHT-mediated transfer of viral resistance was examined. It was found that exosomes purified from PHT conditioned medium attenuated VSV infection in recipient cells (FIG. 1E). The antiviral effect was abrogated using exosome-depleted PHT conditioned medium (FIG. 1E). In addition, exosomes isolated from other cell types, such as an immortalized human placental choriocarcinoma cell line (JEG-3) or primary murine dendritic cells, had no effect on viral infection (FIG. 1E). Taken together, these data point to a direct role for PHT-derived exosomes in the transfer of viral resistance to non-placental recipient cells.

C19MC-Associated miRNAs Confer Viral Resistance

The transfer of RNA and/or miRNAs via exosomes may play an important role in exosome-based intercellular communication (Valadi et al., Nat. Cell. Biol. 9:654-659, 2007; Pegtel et al., Proc. Natl. Acad. Sci. U.S.A. 107:6328-6333, 2010; Zhang et al., Mol. Cell. 39:133-144, 2010). The human C19MC is the largest known miRNA cluster, comprising 46 miRNAs that are highly expressed almost exclusively in the human placenta. Moreover, as a group, C19MC miRNAs are also the most abundant miRNA species in trophoblastic exosomes, with a strong correlation between C19MC miRNA levels in PHT cells and in PHT-derived exosomes (Noguer-Dance et al., Hum. Mol. Genet. 19:3566-3582, 2010; Donker et al., Mol. Hum. Reprod. 18(8):417-424, 2012; Taylor et al., J. Immunol. 176:1534-1542, 2006; Mouillet et al., Placenta. 31:781-784, 2010). To date, the function of these miRNAs has remained elusive. To test whether the expression of C19MC miRNAs could induce viral resistance in non-PHT cells, which do not naturally express these miRNAs, U2OS cells were stably transfected with a BAC that contained the entire human C19MC cluster. When compared to U2OS cells transfected with a control BAC (that is deficient for the C19MC expression sequence), cells stably expressing C19MC-BAC or cells exposed to PHT conditioned media expressed a higher level of C19MC miRNAs, as confirmed by RNAseq (Table 3), and exhibited resistance to VSV infection (FIG. 2A). Likewise, transient transfection of U2OS cells with miRNA mimics of 16 C19MC-associated miRNAs (representing highly expressed miRNAs, or the two subfamilies of the C19MC; Lin et al., Comput. Biol. Chem. 34:232-241, 2010) markedly reduced VSV infection (FIG. 2B and Table 2). It was also found that transfection of mimics of the six highest expressed C19MCs (Donker et al., Mol. Hum. Reprod. 18(8):417-424, 2012; Taylor et al., J. Immunol. 176:1534-1542, 2006; Mouillet et al., Placenta. 31:781-784, 2010) attenuated VSV infection, whereas transfection with mimics of the lowest expressed seven had no significant effect (FIG. 2B). To define the impact of individual miRNAs, individual mimics from among the highest expressed C19MC miRNAs were expressed, and a significant inhibition of VSV infection was detected with mimics of miR-517-3p, -516b-5p, and -512-3p, but not with mimics of several non-C19MC-associated miRNAs (miR-1, -424, -630, and -720; FIGS. 2C-2D) Likewise, a mimic of miR-517-3p also attenuated infection by the DNA viruses VV and HSV-1 (FIG. 2E).


TABLE 2
Groups of mimics to C19MC miRNAs used in the
described experiments. The 16 miRNAs are listed in
order of expression level (highest to lowest)
6 highest
7 lowest
Subgroup 1
Subgroup 2
expressed
expressed
16 C19MC
9 C19MC
7 C19MC
C19MC
C19MC
miRNAs
miRNAs
miRNAs
miRNAs
miRNAs
miR-517-3p
miR-517-3p
miR-517-3p
miR-1323
miR-1323
miR-1323
miR-516b-5p
miR-516b-5p
miR-516b-5p
miR-525-5p
miR-525-5p
miR-525-5p
miR-512-3p
miR-512-3p
miR-512-3p
miR-515-3p
miR-515-3p
miR-515-3p
miR-518e
miR-518e
miR-515-5p
miR-515-5p
miR-517c
miR-517c
miR-519c-3p
miR-519c-3p
miR-519c-3p
miR-520h
miR-520h
miR-520h
miR-519d
miR-519d
miR-519d
miR-518b
miR-518b
miR-518b
miR-512-5p
miR-512-5p
miR-512-5p
miR-520c-3p
miR-520c-3p
miR-520c-3p
miR-518a-5p
miR-518a-5p
miR-518a-5p


TABLE 3
Differences in miRNA expression between U2OS cells exposed to
conditioned or non-conditioned medium, analyzed by RNAseq*
Non-
Fold
C19MC
Conditioned
conditioned
change
Log2 fold
p adjusted
miRNA
medium
medium
(CM/FM)
change
p value
(BH)
miR-517-3p
2374.403
765.215
3.103
1.634
2.66E−32
4.68E−30
miR-519a-3p
1738.835
1144.108
1.520
0.604
9.81E−07
4.60E−05
miR-522-3p
1662.638
1245.641
1.335
0.417
0.001083036
0.021856025
miR-1323
1454.781
459.377
3.167
1.663
8.06E−35
3.31E−32
miR-516a-5p
1093.955
635.203
1.722
0.784
7.75E−09
4.34E−07
miR-521
631.942
494.047
1.279
0.355
0.014010033
0.215579384
miR-1283
565.757
341.747
1.655
0.727
4.06E−06
0.000166442
miR-516b-5p
424.862
188.208
2.257
1.175
1.08E−13
9.49E−12
miR-512-3p
337.442
44.576
7.570
2.920
3.44E−38
2.48E−35
miR-524-5p
269.386
37.146
7.252
2.858
2.06E−33
6.34E−31
miR-515-3p
249.009
17.335
14.365
3.844
4.03E−38
2.48E−35
miR-517c-3p
228.786
116.392
1.966
0.975
2.02E−07
1.03E−05
miR-525-5p
216.140
26.002
8.312
3.055
2.73E−30
4.20E−28
miR-520d-3p
200.409
21.050
9.521
3.251
4.39E−30
6.01E−28
miR-520a-3p
141.669
35.908
3.945
1.980
2.43E−14
2.50E−12
miR-518e-3p
122.653
16.097
7.620
2.930
4.67E−19
5.75E−17
miR-519d
113.300
38.385
2.952
1.562
4.05E−09
2.37E−07
miR-518b
105.055
58.196
1.805
0.852
0.000622055
0.013434194
miR-518c-3p
76.726
32.194
2.383
1.253
2.34E−05
0.000823018
miR-518a-5p
70.166
34.670
2.024
1.017
0.002164332
0.040989113
miR-520g
65.535
61.911
1.059
0.082
0.848007306
1
miR-518e-5p
65.366
26.002
2.514
1.330
5.44E−05
0.001594773
miR-519a-5p
65.366
26.002
2.514
1.330
5.44E−05
0.001594773
miR-519b-5p
65.366
26.002
2.514
1.330
5.44E−05
0.001594773
miR-519c-5p
65.366
26.002
2.514
1.330
5.44E−05
0.001594773
miR-522-5p
65.366
26.002
2.514
1.330
5.44E−05
0.001594773
miR-523-5p
65.366
26.002
2.514
1.330
5.44E−05
0.001594773
miR-524-3p
57.113
2.476
23.063
4.527
1.85E−13
1.42E−11
miR-520h
54.554
34.670
1.574
0.654
0.048469819
0.547397683
miR-526b-5p
48.663
28.479
1.709
0.773
0.020670659
0.277189278
miR-515-5p
43.639
34.670
1.259
0.332
0.311332029
1
miR-498
38.128
17.335
2.199
1.137
0.003592299
0.06503118
miR-527
34.719
17.335
2.003
1.002
0.032552612
0.396755106
miR-526a
33.466
8.667
3.861
1.949
0.000139813
0.00374151
miR-519c-3p
32.153
4.953
6.492
2.699
3.16E−06
0.000138861
miR-520a-5p
26.509
11.144
2.379
1.250
0.014724591
0.223777425
miR-518f-5p
25.322
4.953
5.113
2.354
0.000166051
0.004258507
miR-520d-5p
24.485
3.715
6.591
2.721
4.17E−05
0.001426038
miR-518a-3p
24.118
17.335
1.391
0.476
0.300976907
1
miR-525-3p
20.871
14.859
1.405
0.490
0.270466957
1
miR-519e-5p
17.257
4.953
3.484
1.801
0.008673863
0.14013828
miR-518d-5p
16.733
3.715
4.505
2.171
0.005073798
0.086747848
miR-520c-5p
16.733
3.715
4.505
2.171
0.005073798
0.086747848
miR-518c-5p
14.673
7.429
1.975
0.982
0.150512908
1
miR-523-3p
14.119
3.715
3.801
1.926
0.016108611
0.241825612
miR-518d-3p
11.457
1.238
9.252
3.210
0.006515171
0.108380746
miR-517-5p
9.017
1.238
7.282
2.864
0.018394499
0.263298001
miR-512-5p
8.716
8.667
1.006
0.008
1
1
miR-518f-3p
7.288
2.476
2.943
1.557
0.215985658
1
miR-519b-3p
7.132
4.953
1.440
0.526
0.60445537
1
miR-526b-3p
6.258
1.238
5.054
2.338
0.13991013
1
miR-520e
2.789
3.715
0.751
−0.414
0.91188741
1
miR-519e-3p
1.506
0.000
Inf
Inf
0.579448235
1
miR-520b
1.458
1.238
1.177
0.235
1
1
miR-520c-3p
1.157
0.000
Inf
Inf
0.94380054
1
miR-520f
0.428
0.000
Inf
Inf
1
1
miR-516a-3p
0.000
0.000
NA
NA
NA
NA
miR-516b-3p
0.000
0.000
NA
NA
NA
NA
*Counts were normalized by the median of ratio of the observed counts in each library to the geometric mean of the observed counts of all libraries, as described in Methods. Columns of conditioned and non-conditioned medium represent the mean of the normalized miRNAs counts in those samples.

PHT-Derived Exosomes and C19MC-Associated miRNAs Upregulate Autophagy

Mammalian cells utilize diverse defense mechanisms to combat microbial pathogens. One crucial mechanism is the induction of autophagy, an evolutionarily conserved lysosomal degradation pathway that has been associated with an array of cellular functions. Autophagy also degrades intracellular foreign microbial invaders (a process sometimes referred to as xenophagy) and thus serves as an important cellular response to suppress microbial infections. Exposure of U2OS cells to PHT conditioned medium or to purified PHT-derived exosomes markedly stimulated autophagy, as assessed by the formation of mRFP-LC3b-containing punctae and by electron microscopy, whereas conditioned-medium depleted of PHT-exosomes had no effect (FIGS. 3A-3B and FIG. 7A). In contrast, no effect of PHT conditioned medium or C19MC-miRNAs on type I interferon (IFN) signaling was observed in recipient cells, and antiviral activity of conditioned PHT medium was observed in cells that fail to respond to type I IFNs. In addition, PHT cells themselves also do not exhibit enhanced type I IFN signaling.

Autophagy induction was observed in diverse cell types (FIGS. 3A-3B and FIG. 7B), and was absent in cells exposed to exosome-depleted PHT conditioned medium (FIGS. 3A-3B). In addition, PHT conditioned medium induced the upregulation (>3-fold) of several key pro-autophagy transcripts (e.g., ATG4C, UVRAG, and LC3A) while having no effect on other innate immune pathway components (e.g. toll-like receptors, interferon regulatory factors, cytokine-mediated signaling) in U2OS cells exposed to conditioned PHT medium (FIG. 7C and Table 4), further supporting the induction of autophagy. 3-methyladenine (3-MA), an inhibitor of autophagosome biogenesis, inhibited autophagosome formation in recipient cells exposed to conditioned PHT medium (FIG. 3C). Lastly, incoming VSV particles were trafficked to LC3b-positive punctae formed following exposure of cells to conditioned PHT medium, suggesting that the mislocalization or targeting of incoming viral particles to autophagosomes and/or autolysosome might impact viral replication (FIG. 3D).


TABLE 4
Summary of expression changes in autophagy-related transcripts
Gene
Fold-change
Gene
Fold-change
ATG4C
3.2861
MAP1LC3B
1.2209
UVRAG
3.2696
CHUK
1.2054
CCL2
3.1764
HSPA1A
1.1822
DAPK1
3.119
ATG12
1.1791
CTSS
3.1154
RIPK2
1.1446
EIF2AK2
2.5184
ATG16L1
1.1417
RB1
2.3295
PTGS2
1.1354
BNIP3
2.3171
TOLLIP
1.124
MAP1LC3A
2.1561
EIF2AK3
1.1192
PIK3C3
2.1554
IL8
1.1002
BID
2.149
UBE2N
1.0968
AMBRA1
1.9193
MAP3K7
1.0954
ARSA
1.849
TLR4
1.0872
BCL2L1
1.7855
APP
1.0846
PRKRA
1.7526
PPARA
1.0624
ATG4D
1.7454
PELI1
1.0265
SQSTM1
1.7187
PRKAA2
1.0185
NFKBIA
1.6335
FAS
1.0149
ATG4A
1.6288
BECN1
1.0139
LY96
1.4979
HSP90AA1
1.0107
NFKB1
1.45885
HSPD1
0.9957
TBK1
1.3469
CSF2
0.9879
TP53
1.3446
HGS
0.9745
MAP3K1
1.3294
REL
0.9743
DRAM1
1.3204
CXCR4
0.9529
ATG3
1.289
MAPK14
0.9497
AKT1
1.2812
IRF3
0.9437
TLR3
1.2655
TICAM2
0.9347
ATG16L2
1.2457
ATG4B
0.9261
TGFB1
1.2354
ATG5
0.9159
ULK2
1.2352
HRAS
0.9159
SNCA
1.2286
CTSB
0.9132
HMGB1
0.9027
ATG10
0.724
TNFRSF1A
0.901
FADD
0.7218
ELK1
0.8971
BAX
0.7145
IL1B
0.8966
PIK3R4
0.7073
UBE2V1
0.8948
MAP2K3
0.6992
GABARAP
0.8913
HSPA8
0.6945
IKBKB
0.8859
MAPK8
0.68385
PTEN
0.8814
ATG7
0.6794
EIF4G1
0.8792
MAPK81P3
0.6767
GABARAPL2
0.8772
NFRKB
0.6665
MAP4K4
0.8748
TAB1
0.6663
CD180
0.8699
ULK1
0.6621
CLN3
0.8339
TICAM1
0.6597
PRKAA1
0.8324
CDKN1B
0.6285
FAM176A
0.8304
TGM2
0.6192
TRAF6
0.8257
DRAM2
0.6166
MAP2K4
0.8234
RPS6KB1
0.6072
NR2C2
0.8234
RGS19
0.6005
BCL2
0.8218
NFKB2
0.5946
HTT
0.8135
TMEM74
0.576
TLR6
0.809
ATG9A
0.5758
ECSIT
0.804
CASP8
0.57545
BAD
0.8011
JUN
0.5644
BAK1
0.7987
SARM1
0.5564
MYD88
0.7962
NFKBIL1
0.5528
GAA
0.7936
IRAK1
0.5404
IRF1
0.7825
FOS
0.5091
CASP3
0.7749
TP73
0.5061
GABARAPL1
0.7581
TNFSF10
0.4193
HDAC1
0.7541
IRGM
0.2952
RAB24
0.7351
TNF
0.2475
RELA
0.7325
ATG9B
0.2436
IFNA4
0.1286

Because a role for C19MC-associated miRNAs was observed in the induction of viral resistance, it was assessed whether these miRNAs could induce autophagy. Transfection of cells with mimics of six of the highest expressed C19MC miRNAs (FIGS. 4A-4B), the entire C19MC (FIG. 7D) or mimics of individual C19MC miRNAs that attenuated viral infection (FIG. 4C), also stimulated autophagy, as observed by mRFP-LC3b punctate formation or by electron microscopy. Furthermore, C19MC-associated induction of autophagy occurred via the upregulation of autophagic flux, as supported by a decrease in p62 levels in cells expressing the entire C19MC (FIG. 4D).

The Antiviral Effects of C19MC-Associated miRNAs Require Autophagy

An inhibition of viral replication and a pronounced upregulation of autophagy was observed in cells exposed to PHT conditioned medium and in cells expressing C19MC-associated miRNAs. To determine if the antiviral effects of these conditions involved autophagy, autophagy was suppressed by treatment of cells with 3-MA or by RNA interference (RNAi)-mediated silencing of beclin-1, a key factor in autophagic induction (Liang et al., Nature 402:672-676, 1999). Inhibition of autophagy using 3-MA or by RNAi-mediated silencing of beclin-1 expression significantly restored the level of VSV infection in U2OS cells expressing the entire C19MC (FIGS. 5A-5B). Furthermore, addition of 3-MA to PHT cells enhanced VSV infection (FIG. 5C). These data show that the induction of autophagy is critical for the antiviral effect of C19MC miRNAs.

Discussion

The placenta shields the embryo from the spread of a number of diverse pathogens, including viruses. Disclosed herein is the striking finding that placental trophoblasts transfer viral resistance to non-placental cells. Viral resistance is transferrable via trophoblastic conditioned medium, trophoblastic exosomes, or miRNA members of the C19MC primate-specific cluster. It is shown that multiple members of the C19MC primate-specific miRNA cluster, which is localized to chromosome 19q13.41 and expressed by a specific RNA Pol-II primarily in the placenta (Noguer-Dance et al., Hum Mol Genet 19, 3566-3582, 2010; Bortolin-Cavaille et al., Nucleic Acids Res 37, 3464-3473, 2009; Bentwich et al., Nat Genet 37, 766-770, 2005), are packaged within exosomes, which are capable of carrying their nucleic acids and other types of cargo to neighboring or distal targets (Valadi et al., Nat Cell Biol 9, 654-659, 2007). Although the C19MC is the largest known human miRNA cluster, its function remains unknown. Specific members of the C19MC miRNA family are known to be up-regulated in cancers, such as aggressive primitive neuroectodermal brain tumors (miR-517c and miR-520g) (Li et al., Cancer Cell 16, 533-546, 2009), hepatocellular carcinoma (miR-519d) (Fornari et al., J Pathol, doi: 10.1002/path.3995, Jan. 19, 2012 [Epub]), breast cancer (miR-516-3p,miR-520c) (Foekens et al., Proc Natl Acad Sci USA 105, 13021-13026, 20081; Huang et al., Nat Cell Biol 10, 202-210, 2008), prostate cancer (miR-520c) (Yang et al., Int J Clin Exp Pathol 2, 361-369, 2009), and thyroid adenomas (Rippe et al., PLoS One 5, e9485, 2010). Thus, the data described herein are the first to suggest a unique role of C19MC miRNA members in an antiviral response that is transferable to either neighboring cells within the placenta such as villous fibroblasts, macrophages or fetal endothelial cells, and to maternal systemic cells, such as maternal endothelial or immune cells. Whereas the nature of recipient cells and the mechanisms of targeting remain unknown, exosome-mediated delivery of C19MC family members may constitute a powerful evolutionary adaptation by which a developing fetus is protected from viral invaders during pregnancy.

Primary human trophoblasts produce robust levels of miRNAs throughout pregnancy, as well as other small RNAs (piRNAs, snRNAs, and snoRNAs) (Mouillet et al., Placenta. 31:781-784, 2010; Luo et al., Biol. Reprod. 81:717-729, 2009; Mouillet et al., Birth Defects Res. A. Clin. Mol. Teratol. 91:737-743, 2011; Barad et al., Genome. Res. 14:2486-2494, 2004; Pineles et al. Am. J. Obstet. Gynecol. 196(3):e261-266, 2007). Many of these miRNAs, including members of the C19MC, are found in the maternal blood throughout pregnancy and rapidly decline in the first 24h postpartum (Ng et al., Proc. Natl. Acad. Sci. U.S.A. 100, 4748-4753, 2003; Gilad et al., PLoS One. 3:e3148, 2008), suggesting a miRNA-based mechanism for fetal-maternal communication (Mouillet et al., Placenta. 31:781-784, 2010; Chim et al., Clin. Chem. 54:482-490, 2008). The data disclosed herein thus provide evidence for a novel paracrine and/or systemic function of placental trophoblasts—utilizing exosome-mediated transport of a unique set of primate-specific effector miRNAs to directly communicate with maternal cells, and possibly neighboring placental cells, and regulate their immunity to viral infections. It is possible that PHT-derived, C19MC miRNA-containing exosomes specifically target their cargo to a discrete subpopulation of maternal cells, or may aid in the selectively eliciting antiviral responses and upregulating autophagy. Although placental-derived miRNAs are found in low levels in the fetal circulation, it is possible that these miRNAs are sufficient to regulate specific pathways in the developing fetus, such as the induction of autophagy, which is critical for neonatal survival (Kuma et al., Nature. 432:1032-1036, 2004).

The data disclosed herein show that conditioned media from PHT cells, purified PHT-derived exosomes, and miRNA mimics of several members of the C19MC family potently induce autophagy. Autophagy is an important component of host antimicrobial signaling and often functions to restrict viral replication. Although some of the viruses used in the disclosed study (such as CVB (Schlegel et al., J Virol 70, 6576-6588, 1996; Jackson et al., PLoS Biol 3, e156, 2005) and HCV (Dreux et al., Proc Natl Acad Sci USA 106, 14046-14051, 2009)) are thought to benefit from the formation of autophagic vesicles during their replication, these viruses were also sensitive to the antiviral effects of C19MC miRNAs. Unlike the induction of autophagy via an innate immune pathway in response to virus replication, recipient cells exposed to C19MC miRNAs exhibit robust levels of autophagy prior to their first exposure to viruses. Thus, preexisting C19MC-induced autophagosomes, which fuse with lysosomes to become autophagolysosomes, could profoundly impact the ability of incoming viral particles to properly traffic or release their genomes. It was found that PHT cells themselves also exhibit a high level of baseline autophagy which indicates that this mechanism plays a role in conferring viral resistance to these cells. Alternatively or in addition, autophagy may underlie other important functions of placental trophoblasts, such as those related to feto-placental nutrition or neonatal survival.

Recipient cells exposed to C19MC miRNAs exhibit robust levels of autophagy when first exposed to these viruses versus a typical setting in which autophagy would be an innate immune pathway that is upregulated in response to virus replication. Thus, C19MC miRNAs would greatly enhance the formation of autophagosomes, which fuse with lysosomes to become autophagolysosomes at a very early stage in the virus life cycle, which could have profound impacts on the ability of incoming viral particles to properly traffic and/or release their genomes. The high level of constitutive autophagy in primary human trophoblasts also implies that this mechanism confers viral resistance to these cells. Autophagy may underlie other important functions of placental trophoblasts, related to feto-placental nutrition primarily when resources are scarce. For example, autophagy is critically involved in neonatal survival during the period of starvation that occurs immediately post-birth, when the mother's milk supply has not yet been established (Kuma et al., Nature 432, 1032-1036, 2004).

Unlike the other viruses tested in the studies disclosed herein, conditioned PHT medium and expression of C19MC miRNAs significantly enhanced hCMV infection (FIGS. 6G-6H), indicating that while C19MC miRNAs attenuate the replication of many viruses, they may function in a proviral manner to enhance the infection of CMV, and possibly other viruses. The findings disclosed herein (FIG. 1A) and the work of others (Chan et al., Am. J. Pathol. 161:1371-1381, 2002) suggest that PHT cells are resistant to CMV infection, and studies of CMV-infected placentas suggest that CMV specifically targets invasive and endovascular cytotrophoblasts as a means of entry into the fetal compartment (Chan et al., Am. J. Pathol. 161:1371-1381, 2002; Maidji et al., J. Virol. 81:4701-4712, 2007; Maidji et al., Virology 304:53-69, 2002).

The studies disclosed herein are the first to define an unprecedented role for miRNA members of the C19MC in transferrable autophagy-mediated antiviral responses. The results show that placental-associated C19MC miRNAs are robust inducers of autophagy, a beneficial pathway in states of nutrient deprivation and a powerful suppresser of microbial infections. C19MC-derived placental miRNAs, released into the maternal circulation by exosomes, communicate an antiviral signal to maternal host cells, thus providing an unprecedented mechanism to protect the developing embryo.

Example 3: C19MC MicroRNAs Inhibits HIV Replication

This example demonstrates that PHT-conditioned media, as well as particular miRs encoded by the C19MC, also are capable of inhibiting infection by human immunodeficiency virus (HIV).

TZM-bl cells are HeLa cell derivatives that express high levels of CD4 and the HIV co-receptors CXCR4 and CCR5. These cells are stably transfected with LTR-driven firefly luciferase and LTR-driven β-galactosidase cassettes. Infection of TZM-bl cells with HIV-1 and HIV-2 isolates results in the induction of luciferase and β-galactosidase, allowing for the detection and quantification of infection.

TZM-bl cells were pre-incubated with PHT conditioned medium or control medium for 24 h prior to infection with HIV-1. Cells were then infected with serial dilutions of HIV (0, 3250, 7500, 15,000 and 30,000 pg/ml) for 48 h, and infection levels were assessed by Tat-induced luciferase reporter gene expression, in triplicate. As shown in FIG. 9, PHT-conditioned medium inhibited HIV-1 replication, particularly at the higher doses of virus.

In another experiment, TZM-bl cells were transfected with a scrambled control, miR-517-3p, or miR-516-5p mimics for 48 h prior to infection with HIV-1. Cells were then infected with serial dilutions of HIV (1875, 3750, 7500 and 15,000 pg/ml) for 48 h, and infection levels were assessed by Tat-induced luciferase reporter gene expression. As shown in FIG. 10, both miR-517-3p and miR-516-5p inhibited HIV replication at all dilutions of virus that were tested.

In view of the many possible embodiments to which the principles of the disclosed invention may be applied, it should be recognized that the illustrated embodiments are only preferred examples of the invention and should not be taken as limiting the scope of the invention. Rather, the scope of the invention is defined by the following claims. We therefore claim as our invention all that comes within the scope and spirit of these claims.

<160> NUMBER OF SEQ ID NOS: 82

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<400> SEQENCE: 4

uuucaagcca gggggcguuu uuc 23

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aaagugcuuc cuuuuugagg g 21

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<400> SEQENCE: 6

uucuccaaaa gaaagcacuu ucug 24

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<400> SEQENCE: 7

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<400> SEQENCE: 8

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<400> SEQENCE: 9

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<210> SEQ ID NO: 10

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<400> SEQENCE: 10

aagugcuucc uuuuagaggg uu 22

<210> SEQ ID NO: 11

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<400> SEQENCE: 11

cucuagaggg aagcgcuuuc ug 22

<210> SEQ ID NO: 12

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<213> ORGANISM: Homo sapiens

<400> SEQENCE: 12

aaagugcauc uuuuuagagg au 22

<210> SEQ ID NO: 13

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<213> ORGANISM: Homo sapiens

<400> SEQENCE: 13

ucuacaaagg aaagcgcuuu cu 22

<210> SEQ ID NO: 14

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<213> ORGANISM: Homo sapiens

<400> SEQENCE: 14

cuccagaggg aaguacuuuc u 21

<210> SEQ ID NO: 15

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 15

aaagugcuuc ccuuuggacu gu 22

<210> SEQ ID NO: 16

<211> LENGTH: 23

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 16

cucuugaggg aagcacuuuc ugu 23

<210> SEQ ID NO: 17

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 17

gaaagugcuu ccuuuuagag gc 22

<210> SEQ ID NO: 18

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 18

cucuagaggg aagcgcuuuc ug 22

<210> SEQ ID NO: 19

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 19

aaagugcauc cuuuuagagg uu 22

<210> SEQ ID NO: 20

<211> LENGTH: 21

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 20

cuccagaggg augcacuuuc u 21

<210> SEQ ID NO: 21

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 21

gaaggcgcuu cccuuuagag cg 22

<210> SEQ ID NO: 22

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 22

cucuagaggg aagcgcuuuc ug 22

<210> SEQ ID NO: 23

<211> LENGTH: 23

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 23

gaacgcgcuu cccuauagag ggu 23

<210> SEQ ID NO: 24

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 24

cucuagaggg aagcacuuuc uc 22

<210> SEQ ID NO: 25

<211> LENGTH: 21

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 25

gaaagcgcuu cucuuuagag g 21

<210> SEQ ID NO: 26

<211> LENGTH: 21

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 26

aaagugcuuc cuuuuagagg g 21

<210> SEQ ID NO: 27

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 27

caaagcgcuc cccuuuagag gu 22

<210> SEQ ID NO: 28

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 28

cucuagaggg aagcacuuuc ug 22

<210> SEQ ID NO: 29

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 29

cucuagaggg aagcacuuuc ug 22

<210> SEQ ID NO: 30

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 30

aaagugcuuc cuuuuagagg gu 22

<210> SEQ ID NO: 31

<211> LENGTH: 23

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 31

ucucuggagg gaagcacuuu cug 23

<210> SEQ ID NO: 32

<211> LENGTH: 23

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 32

caaagcgcuu cucuuuagag ugu 23

<210> SEQ ID NO: 33

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 33

cuacaaaggg aagcacuuuc uc 22

<210> SEQ ID NO: 34

<211> LENGTH: 21

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 34

gaaggcgcuu cccuuuggag u 21

<210> SEQ ID NO: 35

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 35

ccucuagaug gaagcacugu cu 22

<210> SEQ ID NO: 36

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 36

aucgugcauc ccuuuagagu gu 22

<210> SEQ ID NO: 37

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 37

caaagugccu cccuuuagag ug 22

<210> SEQ ID NO: 38

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 38

aacgcacuuc ccuuuagagu gu 22

<210> SEQ ID NO: 39

<211> LENGTH: 20

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 39

cuacaaaggg aagcccuuuc 20

<210> SEQ ID NO: 40

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 40

aaagugcuuc ucuuuggugg gu 22

<210> SEQ ID NO: 41

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 41

aucgugcauc ccuuuagagu gu 22

<210> SEQ ID NO: 42

<211> LENGTH: 24

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 42

acaaagugcu ucccuuuaga gugu 24

<210> SEQ ID NO: 43

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 43

aucuggaggu aagaagcacu uu 22

<210> SEQ ID NO: 44

<211> LENGTH: 18

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 44

ugcuuccuuu cagagggu 18

<210> SEQ ID NO: 45

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 45

cucuagaggg aagcgcuuuc ug 22

<210> SEQ ID NO: 46

<211> LENGTH: 21

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 46

aaagcgcuuc ccuucagagu g 21

<210> SEQ ID NO: 47

<211> LENGTH: 20

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 47

cugcaaaggg aagcccuuuc 20

<210> SEQ ID NO: 48

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 48

gaaagcgcuu cccuuugcug ga 22

<210> SEQ ID NO: 49

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 49

cucuagaggg aagcacuuuc ug 22

<210> SEQ ID NO: 50

<211> LENGTH: 21

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 50

caaagcgcuu cccuuuggag c 21

<210> SEQ ID NO: 51

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 51

aucgugcauc cuuuuagagu gu 22

<210> SEQ ID NO: 52

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 52

acaaagugcu ucccuuuaga gu 22

<210> SEQ ID NO: 53

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 53

cucuagaggg aagcgcuuuc ug 22

<210> SEQ ID NO: 54

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 54

aaaaugguuc ccuuuagagu gu 22

<210> SEQ ID NO: 55

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 55

cucuagaggg aagcgcuuuc ug 22

<210> SEQ ID NO: 56

<211> LENGTH: 22

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 56

aaagugcauc cuuuuagagu gu 22

<210> SEQ ID NO: 57

<211> LENGTH: 20

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 57

cugcaaaggg aagcccuuuc 20

<210> SEQ ID NO: 58

<211> LENGTH: 23

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 58

uucucgagga aagaagcacu uuc 23

<210> SEQ ID NO: 59

<211> LENGTH: 18

<212> TYPE: RNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 59

ugcuuccuuu cagagggu 18

<210> SEQ ID NO: 60

<211> LENGTH: 160970

<212> TYPE: DNA

<213> ORGANISM: Homo sapiens

<400> SEQENCE: 60

cctttatggt cagtgtttac agggaagatg gagggaaaat tagagtcaga gattttaggt 60

tttatggcag gatttgggga aaagggattc tggtgtctat ggctcacttt gggaaagaca 120

gtttctagtt tctatggcta gccttggggg agaaggaggg tcaggaagaa acttccgctt 180

ctgaagctgc ttcggaagcc tttgttttga ggttatcggt ttctgagccc caatattagc 240

ctgccacagt ctaagctttc aaagcgcctg tcaaactctt gtggatctat gtatttcaaa 300

gaccaaaatc aacagctcac atatggggtc caagaggagg ggggaggtca ctgcacgttg 360

tcacgtgtga gatctttgtt gttgttgttg agatggaatc tcgctctgtc acccaggctg 420

gagtgcagtg gcaccatctc agctcgactg taaccttcac ctcccaggtt ctagcaatta 480

tcctgcctca gcctcccaag tatctggaat tacgggtgcg tgccatcatg ccccgctaat 540

tttttttttt tttttgtatt tttagtagag acgggggttt caccatgttg gccaggccag 600

tctcaaactc ctgaccccaa gggatctgcc cacctcggcc tcccaaagtg ctgggactac 660

aggcatgagc caccgcgcct ggcctcttgt gagatctgat aagaagtcac gcactagaag 720

gaagggaaga gtgtactcca caaccccaaa cgatgcgttt ggaatcattt caaacccaca 780

taacaagagt ttcatacgga attgggacag aacatggccc ctaatgagcc cccctaaata 840

attctcaagg accacttctc caatgacgaa ggagattgag gatttttttt tcctacgttt 900

attggccgtg tggatatttt gcctttgtaa aatatgtgtc caagcctttt gcccatttct 960

ccagtgggta gtatttcctt tgtttttttt ttttgagaca aagtctcact ctgttgccca 1020

ggctggagtg cagtggcctg atctcggctc actgcaaacc ctgcctcctg ggttcatgcc 1080

agtcttctgc ctcagcctcc caagtgctgg gactacaggt gcacaccacc acgccctgct 1140

aattttttgt atttttagta gagatggggt ttcaccatgt tagctaggat ggtctcaatt 1200

tcctgacctc gtaatctgcc cgcctcggcc tcccaaagtg ctgggattac aggcgtgagc 1260

cactgcacct ggcctaaaag aatctttaaa ttgtccatga gcgatggtgc gtgcctgtgg 1320

tcccagctac ttggaggctg agaaggaagg atcccttgag cctgggagtt tgaggcagca 1380

atgagctatg ctcttaccac tgcactccag cctgggtgaa agaggaagaa gacccttaat 1440

aggcggacca cggtggctca cgcctgtaat ctcaatactt tcagaggcct aggtggaaga 1500

attacttgag accaggagtt caagaccaac ctgggcaaca tagcgagacc ctcatctctc 1560

caaaagtttg aaaaagaaaa aaaaaatttt taatgaaaca gaagtcgtta attttatttt 1620

attttctaag acatcttgtt atagcaaaaa gtccttaatt ttaatatagt ctaatttata 1680

tattttcttc attgtaaaaa tttttgggtc ctatctagca aatgttagct tacctctatg 1740

taataaagac attcttctgt gttgtcttct aaaacaaaac attgttttat tttctgattt 1800

taggtctgca gttcatttgt agatttttgt gtctaggatt aggtagggtt aagatcattt 1860

gtggccaggc atggtggctc acaccagtaa tcctaacact ttgggaggct aaggcaggag 1920

gatctcttga gcccaggagt ttcagaccag cctggggaac atagaaagac cctgtctctc 1980

tatttttttt ttcagttaaa tatatacttt taaaaagatc atttgtggtg ctatagttgt 2040

gggtgtttaa tttgtaattt actttgttct gttgatctat ctaatttttt ttttagacgg 2100

attcttcctc tgtcacccag gctggagtgc ttcagcctct caatgccatc atgcccagct 2160

aattttttgg acttgtagta gagacagagt ttcaccatat tgaccaggct ggtctcgaac 2220

tcctggcctt aggtgacccg cccacctcag cctcccaaaa tgctgggatt ataggcatga 2280

gctactgtgt ccagccagat ctgtctaatt tttttttttc tttgagacgc agtctcactc 2340

tgtcactcag gctggagtgc agtggcgtga tcttggctca atgtaacctc cacctcctgg 2400

gttcaagtga ttttcctacc tcagcctcct gagtagctgg gattacaggt gcgtgccacc 2460

acgcctggct aatttttgta tttttagcag agacggggtt ttgccatgtt ggccaggctg 2520

gtctcaaact cctgacttcc agtgatccgc ccaccttggc ctcccaaaat gctaggatta 2580

caggtgcgag ccactgtgcc tggcccagag ctgtctaatt ttaaagaatg tatgaagatt 2640

cagatttagg ttcctgctat tatgttatgg gaaatctttt cctagtttct tctgattaat 2700

ataaatcctt aattttattt ttatttattt ttttactaca gtcctctttt attacttttt 2760

atttttctta attttaatat aatgaaaagt tcgatttttc ctgttatggt tagcattttt 2820

ttcttctgtt taagaggtgt ttcctggctg ggtgtggtgg ctcacacctg caatcctagc 2880

actttgggag gccaaggtgg gaggatcact tgaacccagg agttcaagac cagcttgggc 2940

aacatagtga aacccttgtc tccaattttt ttaaattaaa aattaaaaaa ggcggggctg 3000

ggtgtggtgg ctcacccctg taatcccagc actttgggac gccgaggcgg gtggatcacc 3060

taaggtcagc agttcgagac cagcctgacc aacatggaga aaccctgtct ctactaaaag 3120

tacaaaatta gctgggcgtg gtggtgcatg tctgtaatcc cagctattcg ggaggctgag 3180

gcaggagaat cgcttgaatc caggaggcgg aggttgcagt gacccgagat tgcgccattg 3240

cactccagcc tgggcaacaa gagtgaaact ctgtctcaaa aaaaataaat aaataaaaaa 3300

taaaataaaa aagaagtgtt tccctagcgt gaagacatga agctcttctg taaacttcat 3360

tgttctgcct ttacaattca atctacaacc cactagaatt aatcttccgt atatatagca 3420

tggggtggag gtcaaacacc ttttttccat atggatgttc agctgtccca gtgtcattta 3480

tttaaaagac tgtactggac ctggcatggt ggctcacgcc tgtaatccca gcactttggg 3540

aggctgaggc gggtggatca cttgaggtca ggagttcgag accagcctaa ccaacatggc 3600

aaaaccccat ctctgctaaa aatacaaaat tagccaggtt tggtggtgca tgcctgtaat 3660

tccggctact tgggaaactg aggcgggaga atcccttgaa cctcataggc agaggttgca 3720

atgagccaag atcgcaccat tgcattccag cctgggcaac aagagcgaaa ctctgtctca 3780

aaaaataaga aaaaatagag gccgagaatg gcccttgctg ccaccaacat ggagactttg 3840

taccgtgtcc cgttcttagc gcttgaatgt cccaacctga agctgaagaa gccgccctgg 3900

ctgcacatgc cgttggccat gactatgtat gctctggtgg tggtgtctta cttcctcatc 3960

accagaggaa tcgtttatga tgttacggtt gaaccgccag gtgttggctc tatgactgat 4020

gaacaagggc atcagaggcc agtagctttc ttggcctaca gagtaagtgg acaatattat 4080

tatggaagga cttggatcca gcttcctgtt tacaatggga ggtttaggtt tcataatcct 4140

ggaccgatcg aatgcaccaa atatcccaaa actcaataga tttcttcttc tattcgttgg 4200

attcgtctgt gtcctattga gtttttttca cggctagagt attcgtgaga atgaaactac 4260

cgggctctct gatgggttag agtgccttta agaagaaatc aggctgggtg cagtggctca 4320

cgcctgtaat cccagcactt tgggaggccg aggcgagcgg attatctgag gtcaggagtt 4380

cgagatcagc ctgggcaaca tggtaaaacc ccatccctac tacaaataca aaattagcca 4440

ggcgtggtga cacatgtctg taatcccagc tactcgggag gctgaggcag gagaattgct 4500

tgaacccggg aggcagagga tgcggtgagc cgagatcgca ccattgcact ccagcctgga 4560

caacaagagt aaatctccgt ctcaccaaaa aaaaaaaaaa gaaaaaaaaa aagaaatcag 4620

tgcatactgg atttgctcct gtcaatgaag ttttaaaggc tgtccaatcc tctaatatga 4680

gatgtagaaa agaaggaaga gcagcagtaa aagaaatatc tagtgaaaaa ccaggaagtg 4740

tattgaagct tggactagaa tttcttcttt attaaagaga caaatttatc acagtatttt 4800

cttttcctgc tgaccacatt gctataccaa tgatgatgag tggcattttc ttcttagttt 4860

tttatttctt taagaaaata caagccaggc gcggtggctc acttctgtag tcccagcact 4920

ttgggaggcc aaggtgggca gatcacgagg tcaggagttc cagaccagcc tggccaccat 4980

gttgaaaccc catctctact aaaaatacaa aaattagccg ggcgtggtgg tggcggggca 5040

cctgtaatcc cagctacttg gaaggctgag gcaggagaat cacgtgaacc tgggaggcgg 5100

aggctgcagt gagcctagat cgctgccact gcactccagc ctgggcgaca gagcgagact 5160

ctgtttcaaa aaaaaaaaaa gaaagaaaga aagaaaagaa aatatactgc atacctacaa 5220

ctataatagc aaatatagtg attatttttt acaaccccct taacactttt tggagatgac 5280

atttctgact ttcagaaatt aacataaaat caagaagcaa gattccatga gctgagaact 5340

ctggacagct ggtcagcttt acctacggag ctttggcttt aactagagtg tgtgatggta 5400

gattatttca gataggtatg taagactgct gcctgaacaa taacatgtat gaaaggaaca 5460

gaaataaata ctaattaaaa aacaaaataa gaaaaaataa aaaataaaca ctactatgga 5520

catggtagct catgcctgta atcctagcac tttgggatgc caaggtgggt ggatcgcttg 5580

aaccaaggag ttcaaggagt tgggagacca gtctgggcaa catagcaaga ccccatctct 5640

aaaaaaaaaa aaaaaaaaag agtactaaag actcagagat agaaaaatgg aaattattca 5700

tattgtctcc agattttttt ttttttggac ggagacttgc tctgtcaccc aggctggagt 5760

gcagtggcac tatcttggct cactgcaacc tccacctccc aggttcaagc aattctcctg 5820

cctcagcctc ccaagtagct gggattacag gcatacgcca ccaaacccgg ctaattttgt 5880

acttttagca gagacagggt tttgccatgt ttgtcaggct ggtcttgaac ccctgacctc 5940

aggtgatcca cccgcctcgg cctcccaaag tgctgggatt acaggtgtga gccaccgcgc 6000

ccagccagta gtcttttatc cttgtaacag attaatcttt tctagggaaa tggagacctt 6060

tcattttcat gtccttttta catccctgtt gattgtatct ctgcctccat tctgttgatc 6120

ggaatgacac taacttaatt tgggttctct aaagcagaca ctgatacaaa ggcttcagtt 6180

ccagaggttt atttagaatg tgatcctaaa ccaagaggga ggtagcaggg agagtaagac 6240

aggaaaggat ggggagccaa caccttatct aaggacatgt tgaggccagg cgtggtggct 6300

catgcctgta atcccagcac tttgggaatc cgaggcgagc agatcacaag gtcgggagtt 6360

tgagaccagc ctggccaaca tagtgaaact ctgtctctac taaaaatgca gaaaacaggc 6420

cgggtgcggt ggctcacgcc tgtaatccta gcactttgtg aggccgagac gggcggatca 6480

cctgaggttg ggagttagag accagcctga gcaacatgga gaaaccctgt ctctactaaa 6540

aatacaaaaa ttagccttgc gtggtggtgc atgcttgtaa tcccagctac ttgggaggct 6600

gaggcaggag aatcacttga acctgggagg cggaggttgc gatgagccga gatcgtgcca 6660

ttgcactcca gcctgggcga caagagtgaa acttcatctc aaaaaaaaaa aaaacaaaaa 6720

acagaaaaaa ttagctgggc ctggtggcag acgcctgtgg tcctagctac tcaggaggca 6780

aaggcaggag aattgcttga acctgggagg tggagattgc agtgagccga gatcatgcca 6840

ctgcactcca gcctgggtga cagagtgaga ctctatctca aaaaaaaaaa aaaaaaagga 6900

taagttgctg aagtctgtga agtggacaat gagggcttga ttcctttgaa gcctgttgaa 6960

gactgtttac gcttcctctt atcatcccct gcctcatccc ctgtcacaga gatgaaagac 7020

tgggacattt ctgtgccaaa tttcatccca cattgggtga ggctttccct gagacatgtt 7080

gacttcctgc agttcaaagc tactttcttc tttagacagg gtctcgctct gtcccccagg 7140

ctagagtgct atagcgtgat ctcgcctcac tgcaagctcc gcctcccggg ttcaagccat 7200

tctcctgcct cagcctcccc agtagctggg actacaggca cctgccacct tgcccggctg 7260

tttttttgta tttttagtag agacagggtt tcaccatgtt agccaggatg gtctcgatct 7320

cctgacctca tgatccgccc accttggcct cccaaagtgc tgacattaca ggtgtgagcc 7380

accgtacccg gcctattttc ttctttagac agggtcttcc tctgtcccct aggctggagt 7440

gcagtggtgt gatcttggct cactgcaacc tctgccccct gggttcaaga gattctcctg 7500

cctcagcctc ctgagcagct gggattacag acatgcgtca ctatgcccgg ctaatttttt 7560

gcattttttg gtagagatgg ggtttcacca tgttggtcag gctggtctcg aactcctggc 7620

ctcaagtgat ctgtctgcct tggcctccca aagtgctggg attacaggtg tgagccaccc 7680

cgcccggggc aaagctattt tccattttaa attttttgtt attgttgttt tgagaccaag 7740

tctcactctg ttgcccaggc tggagtgcag tggcatgatc tctaaatttt gtatttttag 7800

tagagacggg ttttcacctt gttgcccagg ctggtctcga attcctgacc tcaggtgatc 7860

cacttgcctc agccgcccaa agtgctggga ttacaggcgt gggccaccat gcccagcctg 7920

ccaaagctat tttctatggg ataaatgaca gcagaaagga ccccagagca aaatactaca 7980

aagatgtatg gcacgtgcat gaggtgagag tatggtagaa tcaagtgagt ctctgctttc 8040

atgagactga acagtgaggc tcaggttaaa attagaggtg tacaagagag tatgatatgg 8100

acatctgcta ctgaacaatc cttgtgcttc ttgaaaatgc tcatttccat attttttatt 8160

ttttattttt tttcagatac aaggtctcac tgtgtcaccc aggctagatt gcaggggcac 8220

aattaaggtt cactaaagtc ccaacccccc gggttcaagc aatcctcctg cctcagcctc 8280

tggagtagct gggactgcag ttgcatgcca cagtgcccgg ctaatttttt tattttttgt 8340

agaggcaagg gcttgctctg ttgcccagat tggtcttgaa ctcctggcct caaatgatcc 8400

tcccacccgt gcctcccaaa gtgctgggat tacaggcata agccactgca cccagcctca 8460

tttctatatg taagttcatt tagtcattaa gtcttgaagc aaagatccag ccacgatcta 8520

tataaaattt tcaatacagg ccaggcatgg tggctcacgt ctataatccc ggcactttgg 8580

gaggctgagg cagactgatc actttaggtc aggagttcca gaccagcctg gccaacatgg 8640

tgaaacgcta atacaaaaaa ttagctgggc atggtggtca gcacctgtaa tcccagctgc 8700

tagggaggtt gaggcaggag aatcggttga acctggaggt agaggttgca gtaagtcaag 8760

atcacgccac tgcactccac cctgggaaac agagcaagac tctgttccaa aaaaaaacaa 8820

aaggcaatct ttcctagaaa aaaatagttt ctgcacattc ttaggcacgc atttgtaggc 8880

tgctatagct atttaaaatt ttcaatacaa tatgtttacc agagcaaact acaatcctca 8940

ttgttatgtg ggccccaaac cattactgat actcatctgt ttcccccacg tcgtctacat 9000

ttcctcatgg tgggccagta cttcattttt tgtaactcat ttgcctgcta gggtgactca 9060

ggcctttgtt tctgagagat ctgagtctgg tgttgctagg aaggaaaagg ttaactagtc 9120

tataatccat tctttctccc aacctggtga tgtctgagga ggcatccctt ggtcaaactt 9180

ctttgtaaac atgctaatca gatctcatgg tatttattag aggaaaacaa ccagagattc 9240

ttatctgtga gatgctttca gcaggacata cttgttcttt cttgatgtct tgcagtaaac 9300

aaggaattta ggggtttatg ggcatgagcc actggaaaat gtcagcgtca catatttagt 9360

taagagagag agagagaggg gctgggcgtg gtggttcatg cctacaatcc cagcattttg 9420

ggagactgag gcaggaagat cacttgagcc cagaagcgta agatcagcct aggtaacata 9480

atgagacctc gtcttattaa gagtctgatt ctaccctccc ttggtgttat ctcagttact 9540

gtacttttac agggctgtgg ttgtgcaact tcctgctggg catggggaat tctctgagtt 9600

ccagacaaga accttcctgc ccaatatgca gcagcaaccc agtctcttca taatgctcgt 9660

ggttgattat ccatgcccgc caactacctc atttctctgc ccagtgcttt atcagcctga 9720

agagcccaca gtcaccaggc agaaaacctt actgtgcttc ttggtgaaag tgttccctct 9780

tcaaaaatga agacatcacg ccagataagg tttggggact agaagcacaa tttcttcaag 9840

tgggtcccta gacaggatat tgataaatgc caattctgtt tcactccctg gtagctacac 9900

ctgtatattc taactactag ggtcacagcc ccttatattg actgttggtt atatgcagat 9960

attgcatctt agaggacagc accccaaacc catgggatgt tatacctggg ctgacacttt 10020

agttgagccc tttaaaaagc cattcctggg ccaagcccgg tggctcacac ctgtaatccc 10080

agcactttgg gaggctgagg cgggcggatc acgaggtcag gagatcgaga ccatcctggc 10140

taacatggtg gaaccctgta tctaccaaaa atacaaaaaa aattagctgg gcctggtggc 10200

aggcgcctgt agtcccagct attcagaagg ctgaggcagg agaatggcgt gaacccagga 10260

ggtggaggtt gcagtgagct gagactgtgc cactgcactc cagcctgggc gacagagtga 10320

gactctgtct caaaaaaata aataaataaa taaataaata aataaataaa aggccattcc 10380

tggctgggca tggtggctca cgcctgtaat cccagcactt tgggaggccg aggagggcgg 10440

atcacctgag gttgggagtt tgagaccagc ctgaccaaca tggagaaacc ctatttctac 10500

taaaaataca aaattagcct ggcatggtgg cacatgccta taatcccagc tacttgggag 10560

gctgaggaag gagaatcgct tgaatccggg aggcagaggt tgcagtgagc agagattgca 10620

ccattgcact ccaacctggg caacgagagt gaaactccat ctcaaaaaaa acaaaaagcc 10680

attcctggcc tggcgcaatg gctcacgcct gtaatcccaa cactttggga ttacgcttgc 10740

caaagtaagt gaattacctg aggtcaggag tttgacacca gcctggccaa catggtgaaa 10800

ccccgtctct actaaaaata cacaattagc tggacatgat ggcaggcgcc tgtaatccca 10860

gttactcagg aggctgaggg aggagaacct cttgaaccca ggaggcaggg gttgcagtga 10920

gccaagattg cgccattgca ctccagccag ggcaacaaga gtgaaactcc gtctcaaaaa 10980

ataaataaat aaataaataa ataataacca ttatgttatc ccatgatggc agctctttct 11040

gtataaggta ataaatatgc gatgtaaaga gtgtatttca tattaatgca ctgattgttg 11100

tacctttttt tttttttttt tttttttttt gagacagggt cttgctctgt cacccaggct 11160

ggagtgcagt ggcgcaatct cgactcactt caacctctgc ctcccaggtt caagcaatcc 11220

ttctgcctca gcctcctgag tagctgggat cacaggtgcc cgccaccacg cccagctaat 11280

ttttgtactt ttttagtaga gacggggttt tgcctgttgc ccaggctggt ctcaaactcc 11340

tgacctcagg tgatctgccc tccctggcct cccaaagggc tgggattaca ggcacaggga 11400

gccaccatgc ccagctgttt ttttgttttg ttttttgttt tttttgagac agagtctcac 11460

tctgtcgccc aggctagagt gcagtggctt gatcttggct cactgcaacc tcggttcact 11520

gaagcctccc agtttcaagc cgtactctta cctcagcctt ctgagtagtt gggattacag 11580

gcgtgagcca ctgtgcccag ccttttgttt tgtattttta gtagagatgg ggtttcacca 11640

tgttggccag gctggtcttg aactcctggc cacaagtgat ccacctgcct tggcctccca 11700

aagtgctggg attacaagga tgttagaaaa tggctgggcc tggtggctta cacctactcg 11760

ggaaactgag gcacgagaat cgcttgaacc cgggagacgg gcattgcagt gagctgagat 11820

caggccattg cactccagcc tgggcgacag tcacaaaaaa aaaaaaaaag aaagaaagaa 11880

agaaagaaaa cacatcaatt ccaggaaata caaatcagat gagatcgggc gtgttcccgg 11940

tggtatggtc acagaaacag gaagcacaaa tcaatcacct ccaccactgc cctctccggg 12000

ggaaaggatg aaatgtaatc aacttaccac taactagtca gtcattcccc ttaaaggaca 12060

atagcagatc aaggtgtcat catgggcttt tttttttttt tttttgagac agggtctcgc 12120

tctgtcgccc agcctagagt gcagtggcac caccatagct caaccttctg ggctcaagcg 12180

atcctcctgc cttagcctct ggagtactag gactataggt gcatgcctcc atgcctggct 12240

aattgaaaaa ttttttttgt agagacagga tttggctttt ttacccagac tcatctcaaa 12300

ctcctggcct caagcgatcc tcccgcctcg gcctctcact ggtactctta tcacaggcat 12360

gagccagcgt gtctggccct gatcacagcc agaaactccc aggctgtgat aatgagctca 12420

gctttcatga gagacagctc atgctattag gcacatcctt cgtctcttct cctgctaccc 12480

tgctaccgcg tctctgttca tgggtgcatt atacaagcag tggtctgagg atagagcctc 12540

agtgatgccc actgcacaag gcaccctctc ttcctggttg ctcaggatct tctctgtggt 12600

gagcactccc tggagaacat taacatgaga cacagagatc tcatgcctct gcctcttctc 12660

ataattcctt tttttttttt gagagggagt ctcaccgtgt tgcccaggct ggagggcaat 12720

ggtgtgatct cagctcactg caaccttcac cttccaggtt caagtgattc tcctgcctca 12780

gcctcccagg tagctggggc tacaagtgtg tgtcaccgca cccggctaat ttttgtgtct 12840

ttagtagaga cggggtttca ccatgttggc caggctggtc tggaactcct gacctcaggt 12900

gatctgcctg cctcagcctc ccatagtgct gggattacag gtgtgagcca ctgtgcccag 12960

tgccccccct tttttgtgtg tgtcagggtc tcactctgtc acccaggcta gagtgcaggg 13020

gcataatctc ggctcactgc aacatctgcc tccaaggctc aagtggtcct cccacctcag 13080

cctcctgagt aacagggact acaggcacgt gccaccaaac ccagctaatt ttttgtattt 13140

ttggtacaga cgagacttca ccatgttgcc caggctggtc acaaactcct gacctcaagc 13200

gatccaccag ccttggcttc ccaaaatgct gggaatacag acgtgagcca ccgtgactgg 13260

ccccagctaa tttttgtatt tttagtagag atgggttttc accatgttgg ctaggctggt 13320

ctcacactcc tcacctcagg tgatccacct gcctctgcct cccacagtgc tgggattaca 13380

ggcataagcc accacgcccc accgtgagac ccccatctct acaaaaaaag ttttaattag 13440

ctgtgcatgg tggcaggcac cctagtccca gctacctact gaggaggctg atcgcttgag 13500

cccaggaggt caaggctgca gtgagctggg atcacagcac tgcactccag cctgggagac 13560

agagggagac tctatctcca aaaagaaaaa aagactagag attgtgcccc aggcacagat 13620

catattaatc ccagtgatat tctgggagtg gtggccagac tccttcccag gactggtctt 13680

ccggtttggc aatgagtgtt agtcaaggcg tgaggttctc atgggccgtc tcaagcactc 13740

tgttcgacat ctttttactc tctcccaggc tggggtgcag tggcgtcatc ttggcttact 13800

gtagcctccg cctccagggt tcaagcaatt ctggtgcctc agcctcccga gtagctggga 13860

ttataggcgc ctgccatcac gcctggctaa ttttttttgt atttctagta gagtcagggt 13920

ttcactatgt tgcccaggct ggtcttgaac tcctgacctc aggtgatctg cctgcctcga 13980

cctcccaaag tgctggggtt gactgctcgt ggcccctctg ttgcacatct tgacttctca 14040

gcctccaagg tgattcttta ggacaccaat ttaatccctc taaacatggc aacagagcca 14100

gcggtttcca gccctatttg cgcagtagaa ggattattta caaatcctcg tacttagact 14160

ccacccgcag agattcaagt ggactgagcc tggagtgaag ccgaaggcat tttcttataa 14220

tcctgagaat gataatatac agctgtcttt gaaaaccacc actaagactt gtaaactttg 14280

ttcattcaag tcttcttttc tccccttaaa agcaggctta tccttgaggt gtgctttttc 14340

agactccagt aagcatcact gagaacactt aattaatata ctgaggcctc tcacaagagg 14400

gaaggagtgg aagaactcct ggagaaggac ttggaaggga gagatgtttt tctttctttc 14460

tttctttttt ttttttttgg agacagagtt cactcttatt gcccaggctg gagtgcaatg 14520

gcgtgatctt ggctcactgc aacctctgcc tcccaggttc aagcaattct cctgcctcag 14580

cctccccagt agctgggatc acaggcgcct accaccatgc ccagctaatt tttgtacttt 14640

tttagcagag acgggctgtc gccacgttgg cctggctggt ctcgaactcc tgacctcagg 14700

tgatccgccc acctcggcct cccaaagtgc tgggagtaca gacatgagcc accgtgccca 14760

gccaagatgt ttttcttaac acaaaaagtt tttgtctggg tgtggtggct cacgcctgta 14820

ctcccagcac tttgggaggc cgaggcgggg agatcacttg caatcaggag ttcgagacca 14880

gcctggccaa cgtggcaaat acaaaaatta cctgggcatg gtggcgcttg cctgtaatcc 14940

cagttattta ggaggctgag gcaggagaat tgcttgaatc cgggaggcgg aggttgcagt 15000

gagctgagat caagccactg aactccagcc tgggcgacag agtaagactc catctcaaaa 15060

ataaaaaaaa atttctaaag caagcgcttg ggctgttctg atgtcaacat acagtagctg 15120

gccctacctt taacccagga taaggggcaa aagtataccc aaagatacat gtaatcaaag 15180

ttgctgggtg cagtggccca cgcctataat cccagtgatt tgggaggctg aggtgggagg 15240

attacttgat cctggaggtt gagaccagcc tgggtaacac agtgagacct tgtctctgaa 15300

acctaaaata aaataaaata aaataccact gtgctctatc ctgagcaaca gagcaagacc 15360

ctgtctcaaa tataataaaa ataaaaataa tcagtccttg ggtttagcat ctttggacta 15420

atggtaccag catgagggaa agcaggtctt tgtagctctg ataagtgaac tttgacactt 15480

cttagtgttt ttctctaact gtggtgacat acgaataaca tataattaac catttaaaaa 15540

taagcggttc ggtggcattt agtacatcac agtgttgtgc aaccaccacc tctaggtagt 15600

tccaaaactt tttctttctg tttttttttt tgagacagag tttcgctctt cttgcccggg 15660

caggagggca atgacacgat ctccactcac cgccacctct gcctcccagg ttcaaatgat 15720

tgtcctgcct cagcttcccg agtagctggg attacaggca tgtgccaccg cacccggcta 15780

attttgtatt tttagtagag atgaggtttc tccatgttgg ccaggctggt ctcaaattcc 15840

tgacctcagg tgatccaccc gcctcggcct ctcaagtgct gggattacag atgtgagcca 15900

ccatgcctgg ccccaaaaca ctttcctaac tctaaaataa agtcccataa gcaggtattc 15960

cccactccct cctcctcccg gcctgtccat ccacaaatga atggattaaa caatatggtt 16020

tatccataca atggagtata attcagctgt aaaaggggct tggcgcagtg gctcacacca 16080

gtaatctcag cactttggga ggccgaggcg ggcagatcac ttgaggccag gagtttgaga 16140

ccagcctggc caacatggtg aaaccaggtc tctactaaaa atacgaaaat tagccaggcg 16200

tgttagtgca cacctgtaat cccacctact caggagactg aggcatgaga atcacttgaa 16260

tttaggaggc agagaagttg cagtgagctg agatcctgcc actgcactcc agcctgggca 16320

acagagcaag actctgtctc aaaaaaaaaa aaaaaaaaga ggaatgagat accgacacat 16380

tataccatga ggataaacct tgaaaacaac atgctcagta aaagaagcca gcacacaaaa 16440

ggttacatat tatataatgt aattttattt tagtttttga aaagctggtc tcctgggctc 16500

aagcgatcct cccttcttgg cctcccaaac tgctgggatt acaggtgtga gccactgtgt 16560

ccggcctatg attttttttt tttaatgaaa tacctggaaa agataaatcc agagaaacag 16620

acggcagatt acaggctctt tttaaattgg tttctgacac tatcacaaga taaaaatgtg 16680

gtttaaaatt gttgcttggt atgtcttatt cacctctaaa gatatactgt tcatcaggcc 16740

gggtgcagtg gctcacacct gtaatcccag cactttggga ggctgaggcg ggaggatggc 16800

ttgaacccag gagttctagg ctgtaatgtg ctctgccaat tgggcatcca cactaagttc 16860

ggcatcaata tggtgacctc ccaccaggtt gcctaagaag tggggaaatg gtctaggttg 16920

aaaatggagc aggtcaaaat tttcatgctg gctgggcaca gtggctcaca catgtaatcc 16980

cagcactttg ggaggccaag gcagatggat cacctgaggt caggagtttg agaccagtct 17040

ggccaacatg gtgaaaccct gtctctacta aaaatacaaa aaaaaaaatt agctgggcat 17100

ggtggcatgt gcctataatc ccagctattt gggaggctga ggcaggagag ttgcttgaac 17160

ccaggaggca gaggttgcag tgagccggga tcacgccact gcactctaac ctgggcgaca 17220

gagtgagact ccatctcaaa aaaaaaaaaa aaaactttca tgctgatcag tagtggaatc 17280

agcttgtgaa ttgccctcca gcctgagcaa cacagactgc ctcttactaa tcccagcact 17340

ttgggaggct gaggtgggag gatggcttgt gcctaagact ttgagactag cctagccaaa 17400

atcccttctc tacaaaaaat gcaaaaatta gctgggtatg gtagtgtgca cctgtagtcc 17460

cagttactca ggaggcttag gtgagaggct cacctaggcc caggaggtta aggctgcatt 17520

gagctatgat tttgccactg cactccaggc tgggcctagg cctttgtagt cacagctact 17580

tgggggcaaa tgtaggagga tcacttgagc ccagtaggtc gaggctgcag tcagctgtgt 17640

tttttgagac atagcaagac cctgtctcaa aaagaaaaaa aaaagtagct acaagctcat 17700

ttatgcaaag gctaaccact gtcacaagta gagatgtgca gaactgagat tcaaatggat 17760

ggaatggcaa gaaaaactgc cacttctgtg aacctgaagt caaactgccc ttgtcgtcaa 17820

gataaaaggt atacatggtg tgagcctggg catctgaaag gtgttgcagc tttttctttt 17880

atttttttga gatggagtct cactcctgtc acccaggctg gagtgcagtg gtgcgatctc 17940

agctcactgc aacctctgcc tcctgggttc aagcaattct cctgcctcag cctcctgagt 18000

agctgggatt acaggtgccc accaccacac ccagctaatt ttcgtatttt tagtagagat 18060

ggggtttcgc catgttggcc aggctggtct cgaactcctg acctcaggtg atccgcctgc 18120

ctcagcctcc caaagtgctg ggattacagg tgtgagccac ctgtaattag ccaggcttgg 18180

tggtgggtgc ctgtaatccc agctactggg aaggctgagg caggagaatt gcttgaacct 18240

gagaggcgga ggttgctgtg agccgagatc gtgccactgc actccagcct gggtgacaga 18300

gtgagactcc atctctaaat aaataaataa ataaataaaa tgagatgatt tctgagtgag 18360

ttaactaaat caggatatgc agaacaatgc caagcatata ttagccacta aagaatatat 18420

aagttgcctg aaagcccctt tagggtttaa acctggactt cattattcac aatcacattc 18480

ccggtcccca ttacggagcc tggtcacctg ggtgcttgtt agatatggaa atcatcagat 18540

cccaccccag accgagtcag aaacgttgag ggcaggagtt caagaccagc ctggccaaca 18600

tggcggaaac tccgtctcta ctaaaaatac aaaaattagt caggagtggt ggcacaggcc 18660

tgtgatctca gctactcggg aggctgagac atgagaatcc cttgaacctg ggaggcagag 18720

gctgcagtga gccaagatgg tgtcattgcg ctccagcctg ggtgacagag tgggactctg 18780

tctcaaagaa aaagaaaaaa agaaaagaaa ttgcagcttc tgctcaggaa gtctggagtg 18840

ggccaggatt ctgcatttta acaactccca ggagtgttag tggggctggt ttgtggactc 18900

gcctttgagt cgttggtctc cgctcaatca atattagata aaatgacagt attgggagaa 18960

atccaggggg ctctggagga ccgagggatc atattggagg cagatagggc agagaaaggt 19020

ggaggaggtg ggagtcgggt gtggctgtgg aggaagctac ttaaatccgg atttgatctt 19080

tgctagttct tatccctgga cctgaaccca ggcgcacatc tggattagaa gatgccaggc 19140

tcagaggatc ttcgtaaagg taagccagaa aaaatgagaa ccgaagcaaa gacacgtgaa 19200

gaagtggaaa gcagctggcg gcgggaaaag gcagagggac caggcggccg aagctggtgc 19260

ttcgtccacg tgggtggcag ggacttccca cagaggctgt catcgttttt gttgttgttg 19320

ttttgttttg ttttgtttgt cttttgagac agtctcactc tgtcgctcag gctggagtgc 19380

agtggcacga tttcggctca ctgcaacctc cacctcccag gttcaagcaa ttctcctgcc 19440

tcagcctccc gagtagctgg gatgacaggc acacgccacc acacctggct aatttttttt 19500

ttttttgtat ttttagtaga gccgggggtt tcaccatatt gaccaggctg gtctcgaact 19560

cctgaccttg tgatccgtct ggctcagcct cccaaagtgc tggggtgaca ggcgtgagcc 19620

actgcacccg gcccaagttt tgtatcttta tagagatggg gtttcaccat gttagctagg 19680

ttggtctcaa gcccctgacc tcatgtgatt ggctggcctc ggcctcccaa agtgctggga 19740

ttacaggcat gagcctcggt gcccggccgc ggctgtcatc tgacagcaca ggcaaatgca 19800

gggaagcctg tttctgcctt caaatccgaa atcttctccc tcgtagttgt gtcctaattc 19860

ttcccacccc gatgcttgtt cttgagcaca ggggaatctg attcctatga ggatgattag 19920

gtcctagata gaatataggc ttagccaagc tggaaatctc atgttcatga gcacttacta 19980

gactctggag ctgctgccaa accattttca ggcatgatgc gattcagatc ttttagagcg 20040

aggcatgcgg tggctcatgc ctgtaatccc agcactttgg gaggccgagg tgggcagatc 20100

gcttcagccc aggagttcaa gaccagcctg ggcaacaaaa caagacgttg tctctacaaa 20160

aaatacaaaa attagccggg catgtcagtg ttgtgtagtt ccagctactc gggaggctga 20220

ggtgagagga tcgcttgagc ctggggaggt tgaggctgaa gtgagctgtg atcatgccac 20280

tgggcaacac agcagtacac tgtctcaaga aaaagaaagt aaaaggtgac atccgtgaca 20340

gcagtgatgt tatttttctg cttatttcct tgctatattc ctaaaagaat atacttaatg 20400

aacattgaat aaatgataag aataaaacca tgtccatcca tttcaccttg gattgagttg 20460

cctatttcta cacaaagaaa atagaaccca agaggtaagg cacaagcgat cccttattta 20520

cgtatttatt tatttattta tttattattt tcttgagaca gggtctccct ctgtctcagg 20580

ctggagtgca gtggcataat ctcggctcac cacagcctct gcctctgagg ttcaagcgat 20640

tctcctgcct cagcctcccg agtagctggg actacaggcg tgcaccatca cgcccagcta 20700

atttttgtac ttttagtaga gacagggttt caccatgtta gccagaatgg tcttgatctc 20760

ttgacctcat gatccgcccg ccttggcctc ccaaagtgct gggattacag gcatgagcca 20820

ctgtgcccgg ccaaattttt gtgtttttaa tagagatggg gtttcaccat gctggccagg 20880

ctggtctcaa actcatgacc tgatatgatc tgcccgccct ggcctcccaa agtgctggga 20940

ttacaggcgt gagcaaggga tcccatattt aaatgataac agaaaaaaag gatggaggaa 21000

ccaaggggga aaggaacaac cttttcctat gaaaatgaca aatgaggctt ggaaaaacaa 21060

ggacaaaagg cagatcaagt tgtcctcccg cttcagcctc ccacagtact gggactgcag 21120

cctgaggcac gaccccggcc agcagtgtct ccatatctaa gaacctcagt cacggccggg 21180

cacagtggct cacgcatgta atcccaacac tttgggaggc cgaggcgggt ggatcacctg 21240

aggtcaggag ttcgagacca gcctagccaa catggtgaaa ccctgtctct gctagaaata 21300

caaaaattat ccgggcatgg tggcgtgcgc ctgtaatccc agctactcgg gaggctgagg 21360

caggagaatc gcttgaactc aggaggtgga ggttgcagtg agctgagatt gtgccactgc 21420

actccagcct cggtgacaga acctcagtca cttgatcatc acgttgtacc tcagtggaaa 21480

ggaaaggaaa gctcaggctt ttgagaatgg agttgttagc atttcccatt tgtgtctttt 21540

tcctcctctt tcaaggcaag gaccagatgc attcacacag gaaacgaacc atgttcacta 21600

agaagcaact ggaagatctg aacatcttgt tcaatgagaa cccataccca aaccccagcc 21660

ttcagaaaga aatggcctcg aaaatagaca tacacccaac agtactgcag gttggaaaat 21720

gatccctctt ctcactaaac tgccttcctg atctaatcta aattcagagt ccctctagga 21780

taattcctga ggtctcattc caatcgccaa tattccccaa acccacactc tcctactcac 21840

gtccccgtaa acctttcttc aaccccctag agcaagaatg ggaaaatttt tccagtaaag 21900

gaccaggtag taaataattg aagctttgtg ggccataccg tctctctagc aactatttta 21960

actatgccac cacagtgcga aggcagccac agacaacgta aacaagaggc ccatctgtgt 22020

tcccagaaac tcttttactc aggttggagc acggtggtgc aatcacagct cactgcagcc 22080

tcgacctccc gggctcaagc aatcctacca cctcagcacc ccaagtagct gggactacag 22140

gtctacaggt gcacactacg cctcccaaag tgttgggatt acaagcatga gcccggctgt 22200

attttatttt ttgtagagat ggggtttcgc tatattgccc aggaggctga tcttgaactc 22260

ctggtctcaa gtgattcacc tgccttggcc tcccaaagtg tcaggattat aggcataagc 22320

cactgcgcct ggcctgaagt ttatacaatt ttcacattac aaaatagtgt tgttttgttt 22380

tgttttaaga cagggtgttg ctcttgccca cattggagtg cagtggcacg atcatagctc 22440

actgcagctt tgaattcctg ggctgaagcc atcctcccac tcagattccc aagtaggtgg 22500

gactacaggt gtgtggtact acagcctccc gagcagctga tactactggt gtgtaccacc 22560

aggcccagcc aatttgtgtg ttttttgtag agatggggtt tcgtcatgtt gcccaggctg 22620

gactcaaact cctgaaatca agtgatccac ccacctcagc ctcccagagt cctgggatta 22680

taggcatgag ccactgtgcc tggccagtag tattctttta ttttctctct tttttctttc 22740

agtcccaaca cacatacaac acaaaatagt actcttattg tgtttgtttt ttctcaacca 22800

ttttaaaacg taaagcctat tcttagtgca tgggtgatac aaagacaggg ggtggtcaga 22860

tttggccacc agccttagct ggctggcctg agccccagag cagcccctgc gactgatccc 22920

cttgctctcc tatcccctgc tctgggtttt ctgacccctg tctcaatttc tgtccccaaa 22980

atctctcatt tctgctcctt cctgggagta gattgagtag ggttccacaa agaggatgta 23040

agtggccaag ctgtggcact agctgttctc acctagaagt actcatatta tcaactaaaa 23100

ggaaaacttg ggttgggtat ggtggctcat gcctgtaatc ccagcagttt gggaggccga 23160

ggtgggcgga tcatgaggtc aggagttcga gaccagcctg gccaatatgg tgaaaccctg 23220

tctgtactaa aaatacaaaa attagccagg catagtggtg tgagcctgta gtccccgcta 23280

ctcaggaggc tgaggcagaa gcatcgcttg aacctgggag gcggaggttg cagtgagccg 23340

agatggtgcc actgaactcc agcctgggcg acaaggtgag actccatctc aaaaataaaa 23400

taaaataaaa taaaaataaa taaataaata aatattagga gagtttaagg gccaaaaaaa 23460

ggaaaaaaaa agaaaagtta aaaacttaaa ggttaaaaaa aagaatataa ataaataaaa 23520

tacgtttaac aaaataaaaa ttaaaaagta aaaatttttt aaaaatagaa tgacaagtca 23580

tctgaattcc accccattct cttctctctc ttcccttcag gtctggttca agaatcacag 23640

agcaaaactc aagaaagcga aatgcaagca tattcatcaa aaacaagaaa ctccacaacc 23700

gccaatacca gagggtgggg tctccaccag tgtcggcctg agaaatgcag acacactacc 23760

cagattgccc aacgctgctc acccgatcgg cctggtgtac acgggtcatc gagtcccctc 23820

attccagctc atcctgtacc ccaacctcaa ggtccctgca aatgacttca ttggccacag 23880

aatagtccat tttggctgct gccgagatcc taatatatac tgcctctacc ccattttgga 23940

atcccaagtt tgcgctccaa gcttccattc tggctctcct gcctgttcat ctaaccaaag 24000

tcgagagaga tgataaatac aaaaagtcac atgttgtaat gatgtgtgtg tggtactgtg 24060

acatttgcgt ttggtcttcg tgcctgtttc ctggaaagca gctccagagt tccatggagt 24120

ctccaaagtg ccatcttttt gtattttgtg tgctaatgtt gaccgatagc ttcaggatgg 24180

ggactggtct ctggaaagac cccagtagga ttagagggca aacttctggg aggggaagga 24240

agctgagggt gaggctgatc accattggcc agtggtttca tcagtcatgt ctgggtaagg 24300

aagcatccat caaaacccag gaggacaggg tttgaagaga tccctgacag ctaagcatgc 24360

ggacgtacct ggagggtggc atatccgggg agggcatgga agctccgggc ccctcgtcat 24420

acgcctcatc ctagcagggt gcggtggctc atgcctctaa tcccagcacg ttgggaggcc 24480

aaggcaggcg gatcacctga ggtcaggagt ttgggaccag cctggccagc atggcgaagc 24540

ccatctctac taaaagtaca aaaattagcc gggcgtggtg gcttgtgcct gtaatcccag 24600

ctactcggga ggctgaggca ggaggatggc gtgaacccgg gaggcagagg ttgcagtgag 24660

ctgaggggat tgtgccactg cattccagcc tgggtgacag agcaagaatc tgtctcaaaa 24720

aaaagaaaca agaaagcaca gaggaccagc tggaatattt ggaagaccag aaaatgtcct 24780

taaaactcaa aatgcctggg catataggtc aggtccacct agccttttga atcaatgcga 24840

acccttcctc aaggcctgga ggttggcaac acaaaatcac cctgtggaga acctaaagca 24900

ttccctctct gagagttgag tcaattggga ttccaccccc aaaattccta aaaatctcaa 24960

gttcctagaa aattctcaga aaaagcaagc tctcctatac ctccctggct gtactctgat 25020

gcaacttggc aaccatcaga gaatctccat ccccttgaac cttggtcctc tgccaatgtt 25080

cctcacgcct gtaatcccag cactttggga ggctgaggcg ggcggatggc ttgagcccag 25140

gagttcgaga ccagcctggc cagcatggta aaacctcatc tctaccaaaa acatcaaaat 25200

tagccagtct catatactgg tctcaaaata aataaataga ttaaaattta aacataaaat 25260

aaacatgagt gtgttagaaa gaacctactg gcccggcgtg gtggctcaca cctgtaatcc 25320

cagcactttg ggaggcagag gcaggcggat catctgaggt tgggagttcg agaccagccc 25380

gaccaacatg gagaaacccc gtctttacta aaaatacaaa attagccagg catggtggcg 25440

catgcctgta atcccagcta ctggggaggc tgaggcagga gaattgcttg aacccgggaa 25500

ggggaggttg ccatgagccg agattgcgcc attgcactcc aagcctcctg ggcaagaaga 25560

gcaaaactcc atctcaaaaa aaaaaaaaaa aaaaaggaac ctactgtagg atttggagtt 25620

gagtaatttc agatagaatc aaaggaagag aaaggtctgg attgggtggt gccagcggca 25680

gcttctatga ctgagtatct caatacatct tatctatagg acggctgact agaaagagga 25740

caaacctata atagataaat gggcggcagc cattgcttag ccataggcga atatttgatg 25800

ggttggggct caggacaaat gttaaaaaca atcttaataa cagcatgccg gcccggcggc 25860

ccacacctgt aatcccaggg gatttcagag gtggaggggg aggcggaagg attgcttgag 25920

cccaggaggt tgaggctgca gtgagctgtg attgcaccgc tgcactccag cctgggtggc 25980

agagcgagcc cctatatcaa aacaaacatt atcaagaaaa attattaaat gaataacaaa 26040

aacaggctgg gcacggtggc tcacgcctgt aatccaagta ctttgggagg ctgaggcagg 26100

tggattgctt caactcagga gtttgagacc agcctaggca acacggtgaa accctgtccc 26160

tacaaaaaca caaagaagga gtctcgcttt gtcgcccagg ctggagtaca gtggtgtgat 26220

ctcggctcac tgcaacctcc tccatctcct gggttcaagt gattctcctg cctcagcctc 26280

ccaagtagct gggactacag gcatgcacca gcacacctgg ctaatttttg tatttttagt 26340

agagatgggg tttcatcatg ttggccaggc tggtctcaaa ctcctgacct caggtgatcc 26400

acctgcttca gcctcccaaa gtgctggaat tacaggcttg agccaccatg cccagcctct 26460

acaattaatt aaaaaagaac cataacccca aaacttgtac aaccaccagt ccaagaaacc 26520

aagtcacaat ttctgcagca atcagcccca aatgcccagg gcttgatcca tcactcgtag 26580

cttccctagt ttttgtgccc atcccccaac cttgatttca atttaggacc aaccagagaa 26640

acccaatgtt tggctaaaca atcccacagg atcctacttc tggttagctc cctggcggcc 26700

tcccaacgtc agcagccttt ggtcaggaca tatctgaaga cttcgttttt tttctgctat 26760

aaagctttct ttcctttttt ttttttctcc tgaggtgaag tctaactctg tctcccaggc 26820

tggaatgtag tggcatgatc tcggctcact gcaacctccg cctccctggt tcaagcgagt 26880

ctcacacctc agcctcctga gtagctgaga ttacaggcat gcaccaccac acctggctaa 26940

tttttatggt gttttttttt gtttgttttt ttgtttcttt ttgtagagac ggggtttcac 27000

catgttgccc aggctggtct cgaactccta acctcaaatg ttctgcccaa ctcggcctcc 27060

caaagtgctg ggataacagg cgtgagccac tgcgcctggc catggaacct cactctttaa 27120

agtgtgcagg gaccccagtc attccccact gtgtcgcggt tcccataaca gtgcctgtca 27180

ccaagtatgt tttcaaaata catttttgga agagagagag aaacagaagg ggagggggta 27240

ggaagagaaa gtaggggaag aggaaagtga gagagggaga caggaaggga gagggaaggg 27300

agacacagag atgaagacaa agggaaagag acagaggccg ggtacagtgg ctcatgcctg 27360

taatcccagc actttgggag gccgaggtgg cggatcacct gaggtcagca gttccagacc 27420

agcctggcca acatggtgaa acccccgtct ctaccaaaaa tacaaaaatt atctgggagt 27480

gatggtgtgt gcctgtaatc ccagctactc gggaggctga ggcaagagaa tcacttgaac 27540

ctgggaggtg gaggttgtgg tgagccgaga tcgcaccctt gaactccatc ctgggcaaca 27600

gaacgaggct ctatctcaaa aaaaaaaaaa aaaacagaaa gaaaagaaaa gaaaaggaac 27660

agcttccaag catctttgga gaagggcttc actaacatca aatcctgtct tcctctcatc 27720

accttctagc ccagtatttc cccaacgtga gaataataag aatcatctgg ggagctgtta 27780

aaacccagac tctgagacca gacaggggca tgcttcccag gtgtgggcat gatttttccc 27840

tggggttgta tttaacaaag ttcaaaaccc aggaggtaca atcaaaacct gagccgtctg 27900

gagactcaag acttcttagg gaagacgaaa aagagaaatt ccggccaggt gcagtggctc 27960

tcgcccgtaa tcccagcact ttgggagtcc aaggcaggag gatcgcgtga tcccaggagt 28020

tggagaccag cctggccaac atggtgagac cctcatctct acaaaaaaaa aaaaaaaaaa 28080

agagagagag agagaaagaa gttcccagag aaagttccca gaggaaacag gagggaaaag 28140

ggcgaggact gcacccagcc agctagggta tgagtgtgaa ccatagtcta gggggctggg 28200

tgtgaattcc cagtccaggg gctgtgaata cgtggagaca ccgatgctgg ccctgcaccc 28260

agcctcctcc ctgggcgaac tattgactga agccacccct cgtgtcccca ctgttcctgg 28320

tctcaacttc ctcacctgat gaatgggcgg ggagggtaat tcccctacgg tggggctttt 28380

ctcactctgt tatgtgttct tacaaaatgt gctctctgat atcatctata tctccctaga 28440

aggaatcatt agtttatctt tttttttttt tgtgggacag ggtttcactc tggtcgccca 28500

ggccggagtg cagtggtgcg atctcggctc actgtaacct ccacctcctg ggttcaagcg 28560

attctcccac ctcagtctcc caaggcctgg tgcggtggct tacgcctgta atcccagcac 28620

tttgggaggc caaggtgggc ggatcacttg gggtcaggag tttgagacca gcctggccaa 28680

tatggtgaaa ctccctcttt actaataata caaaaattgg ccaggcgtgg tggcgggcag 28740

gtgtaatccc agctactcgg aaggctgagg caggagaatt gcttgaaccc agaaggcgga 28800

ggttgcagtg agccgagatc gtgccactgc actccagcct ggggacagag tgaaactgtg 28860

tctcaaataa taataataat aataataata ataatttagg ccaggtgcag tggctcacgc 28920

ctgtaaaccc agcactttgg gaggccgagg cgggtggatc acgaggtcag gagttcaaga 28980

ccagcctggc taacatggtg aaaccccatc tcaactaaat atacaaaaaa attagccggg 29040

cgtggtggca catgcctgta atcccagcta cttgggaggc tgaagcagga gaatcgcttg 29100

aacccaggag ggggaggttg cagtgagcca agaccgtgct actgcactcc agcctggcaa 29160

cagagcaaga ctccgtctca aataataata ataataataa tttaaaaaaa aaataaaaaa 29220

taagaaataa aagaatgaac cacaatgtcc ctttgttcta caggccactc ttgtgctatc 29280

cacagttgcc aaaaaccccc acctgccttg tctatggacg cccatcgggt gataccacaa 29340

acacctttga tcctacagat accagggtaa attcagcatc caagtccttg aattcttccc 29400

gtgagtataa gctgccctgg ctcagccctc ttctttgcca ggaacccaat ccttcaaact 29460

tccctcaaag actgggtact tttagcatga ctgctctaga gattctgggt ggggctcaaa 29520

gtttaacctt cagggtcagc aacattttat ggaaagagcc acagagtaaa tatttgaaac 29580

attctcgtac actgacactc agctggactg gcaaagcaga atatctgtgt gtcagtgtgc 29640

gttttattca gccgttgttt gggtcagggt ctgtgggcgg cccctggcag ctaatgccct 29700

cctgtgagga acaatacctc accatcaagc acagtccgtc acatattctt gccctggtcg 29760

ctcgctcttt ctttcttttc tttctcttct ttctttcctt tctttctcat tttcgtttct 29820

ttctttcttt ccttcttttc tttttctttc tgtctctctt tatctatttc tgtttctttc 29880

tctctctctc acctttcttt ctttccttcc ttccttcttt ctcttccttt ctttctctat 29940

ctctctttct ttctttcctt ccttctttcc ttgtttcctt ctttcttttc ttttctttct 30000

ttttctttct ttcttttctt tctttctctc tctctctttc cttttctttc tctctctctc 30060

tctctctttc tctctttctt tctttctgac agggtcttgc tgtgtcaccc aggctggaat 30120

gccaagatgc actcataggt cactgcagcc caaataaatt tccccaggct caggtgatcc 30180

tcccacctca cccttccaag tagctaggac caacaggtac atgccaccat gcacgtctaa 30240

tttttatttt tattttttga gactggtctt gctctgtcac ccaggctgga atgcagtggc 30300

acgatcacag ctcactgcag cctcgacaac ccggcccctc caagcaatct cccccctcac 30360

cctcccaagt agctgggacc tcaggcatgt accaccatgc cctgctaatt ttttattgtt 30420

tgtagaggcg gggtctccct atgttgcgca gggtggtctt gaacacctgg cctcaagcaa 30480

tcctcctgcc ttggccaccc aaaatgctgg gatcatgggc gtgagccact gtgcctggtc 30540

tggtcttcct cctttcatcc tcttagtcct tgacctgtcc ctcttctaga gtcctccctc 30600

ttcatgaccc tctccccctt gttggggctc agaaactgat acctctaaat acagcatgtt 30660

aaaggtgcct caagggccgg gtgcagtggc tcatgcctgt aatcccagca ctttgggagg 30720

ccgaggcagg cagatcactt gaggtcagga gttcaaggcc agaccgacca acatgcaaaa 30780

ccctgtctct actaaaaata tggaaaaaga aaattagttg ggtgctgtga cacacgcctg 30840

taatcccagc tacttgggag gctgaggcag gagaatctgg gaggcggagg ttgcagtgag 30900

ccgaggtcac accactgcac tccagcctgg gcaacagagc gagactctgt ctcaaaaaat 30960

aataagaaat aaataaatga gcctcaaggt ctctgacctc ccccgatcat ctcttccaaa 31020

gcacaggcct ttatctgtgt aagatccaga cctgccaaga ggaataattg ttttttcttc 31080

tcctccctgt gagacaaaga atgtaaccac acctgaacag atcccatcac tgtcaatagg 31140

aatgatttcc aggacccatt catgctctct agtaatcctt tattgccctc aatggaattc 31200

ctctagcccc ctcccataat cagtttgcca ggagaacata taagcttctg aacccccttg 31260

ggggatgggc gatcactggc tctccccatg ctcaggttaa agacatttgt aagccttttc 31320

tcctgctaat ctgcttccta tcagttcttt tcagccaacc ttcagagagc caaaggcagc 31380

cttcaccttg gcctcaacac cttccatgcc ttttgcttct cctcctcccc tgcaaagcat 31440

tccccactcc ttgggttccc cactttcaag catcccaccc tccctgttat atgtcatcct 31500

cctcattctc tgcttttata acccttttaa aattatgggc ccagccaggt gcagtgatgc 31560

atgcctgtaa tcccagctct ttgggaggct gaggcaggca gttcacttga gctcaggagt 31620

cctacacagc tggggcaaca tggtgagacc cccatctcta ccaaattgca aatattagcc 31680

gggtgtggtg gtgtgagcct gtattcccag ctactcagga ggctgaggta ggagaatggt 31740

ttgaccccag gagatggaag atgtagtgag ccaagatcac gacactgcac tccagcctgg 31800

gcaacagagc aagatcgagc ctaaaaaaaa aaaaattagg tggggtgcgg tggctcacac 31860

ctgtaatccc agcactttgg gaggtcgagg caggtggaac acctgaggtc aggagtttga 31920

gaccagcctg gccaacatgg caaaacccca tctctactaa aaatacaaaa aaattagctg 31980

agtgtggtgg cgggtgcctg taatcctggc tacttgggag gctgaggcag gagaatcact 32040

tcaacctggg agccagaagt tgcagtgatc tgagataatg ccattgcact ccagcctggg 32100

tgatagagtg agactctatg tcaaaaaaaa aatacataaa aaaattaaaa attaaaaatc 32160

aatttaaaaa atgccagcta gtatactttt cctctctccc ttcctttttc tatctctctc 32220

tctttttttt tttttttttt tttttttttt tttttttgag acagtttcac tcttgtcgcc 32280

caggctggag tgcaatggcg tggggatctt ggctcactgc aacctccacc tcctacactc 32340

aaatgatcct tctgccttag cctcctaagt agctgggatt acagatacaa gcaacaaatc 32400

ctaattcttg tatttttcgt agaggcaggg ttttgccatg ttctccaggc tggtctcaaa 32460

gcgaatgctg gccaggctca tttttttaat ttgaattgtg ggaaaactga ggctggatcg 32520

aatggtcctg gacatgccct ggaacaaaca acatttagct gtgcaaggca cagtgtcagt 32580

gtctggtatt ggggtgcccc agtctcaagt cccaagaaaa agacaaggca gaccatgaat 32640

tttgtgtggc tccaactcag ttttcttcat ctgttgcttc acttgcagtg ccagaccaga 32700

atcactttga cctaaaacta gctacgataa ttaacattca gtatgactca ttacctaaca 32760

aattcaaaat gtaaggaact gtctttagaa cttgttctat ataaacatgt tattaaggtc 32820

tgctgcggct gttctatgca gctctttttt ttttttttct ggcgatagga tctcactcta 32880

tcgcccaagc tggagagcag tggcatgatc acagctcact gcagcctcaa cctcctgggc 32940

tcaagcaatc ctcccacctc agcctcccaa agtgctggga ctacaggcgt gcacccctgc 33000

acccaactag ttttgtattt tttgtagaga tggagtgtca ctatgttgcc caggctggtc 33060

tccaactcct ggcctcaagc aatccaccaa ctcggcctcc caagtgctgg gattacaggc 33120

ggacgccact gctcccagcc ccagattttt ttctcattta tatttccgca aacctcaaaa 33180

aaacccccaa aggtccctag gtcagtcact tcccactagt aaataagtca gcatctttgc 33240

aaagatcaaa ggaaaagcaa ttttccttac cacagagcct cttacaggaa agagagctct 33300

gattttccat atgttcccca accatgctag gatgttcctg tgccctgatg gcattcttac 33360

taacccattg ttccattcct ttccagccct ccttcctgta acgtaagtgc aggtgggagt 33420

tggggattgg tccgctaact gggtatagag aaaacagatg ggtatgctat tctcagtaag 33480

ggagcttctg tcctgggaaa aatgtggcct ctcctaagga accctatccc attgaatcac 33540

cctgattcac tttttttttt tttttttttt tttttttttg agatggagtc ttgctctgtt 33600

gcctaggctg gagtgcaatg gtgtgatctc ggctcactgc aacctctgtc tcccggttct 33660

agtgattctc ctgcctcagc ctcccaagta gctgggacta taggcataca ccaccaagcc 33720

tggctaattt tttttttttt tttttttttt tttgagacgg agtcttactc tgtgccaggc 33780

tggagtatag tggcgtgatc ccggcccgct ggctcactgt aacctccgcc tcctggattc 33840

aagtgatttt cttgcctcag cctcttgaat agctgggact ataggcgccc accaccacac 33900

ccagctaatt tttgtatttt tagtagaaac aggttttcac catgttggcc aggatggtct 33960

cgatctcttg acctcgtgat ccacacgcct tggcctccca aagttctggg attacaggcg 34020

tgagccactg cgcccggccc taatttttgt gtttttagta gagatggggt tttgccatgt 34080

tggccaggct ggtctggaac tcctggcctt aggtgatctg cccacattgg tcttccaaag 34140

tggcgagatt acaggcaaga gccactgtgc cctgcctctc acattttctc tcttcccaga 34200

tttggttcaa gaacttaaga gctaggccat gtgcggtggc tcacgcctgt aatcctagca 34260

ctttgagagg ccaaggcagg cagatcgctt gaactcaaga gttccagacc accctgggca 34320

acatggtgag atccctgact acaaaaattc gccaggtgtg gtggcatgtg tctgtagtcc 34380

ctgctattca ggaagctgag gcaggcggat cgcttcagcc tgggaggtca agactgcagt 34440

gagctgagat ggagacacag cactccagcc tgagcgacag agccagaccc tgcctcaaga 34500

aaacaaaaca aaacgataac actggagagc cagagcagga accagtctcc tctccaatct 34560

ccaaacctga gagagacatg agtcacactg acggctttgc ctgcctgcga tctctctatg 34620

tgtgttgatt cttgcggaac aaggcgcttc gagcatctcc tctgccccta cgcccgtgac 34680

cctgtccctc ttcttgtctc cattgccccc caagcgggta catttgccca ccttctcacc 34740

tgggacaccc tgggcgtgga tctcctcacc tgcagcgcta ggtgtgctcc cagggtctcc 34800

acatccctaa cccccgcaag gctggcctct ttacctgcaa agccccccgc cgcggcctcc 34860

tccctctacc ataccccaat gccaggctca cttcctgccg cctgcctgga acggggctgc 34920

tcatgcatcc cccacgccct ctgaagcccc cccggcgcac tccacgccct ctcgcctgac 34980

ccctgtttcc gctgccggcg tctccacacc ccctgacgcc gccacgccct ggaccgaggt 35040

ctctagagct gcgcgccggc tgcacgtccc ttaggagttt ccgtgcgcca cgaggcgctg 35100

gcgcgcgtct cccggcccat ccaccccggg cccggcgaca cctttctttg ccacctggaa 35160

ccaacatctt ggttcccttt tgagggatca gaacttgttt aattggaata cggcaaaatt 35220

ctgaattttt ctgccgtctc tattccaact tcagagttct gccgtccagc cctgcgacaa 35280

tcttccggtg ccaacgcggc aggtcagtat gtatccccca cgatgccccc cgggccacgg 35340

gcccctagtt aacaggtttc cctttcgccc gctgcctgga agtatcgcca cctcgccccg 35400

cccaaccccc caccagacag ctctgcagcc acagcccctc atccaaccag gaagtccagg 35460

gcccatctgg cccgctagac ctcgggaaac cacggcgtca gagcacccat taagaggggt 35520

ccaggccggg cgcggtggtt cgcatcaggg cgcccattaa gaggggtcca ggctgggcac 35580

ggtggttccc atcagggcgc ccattaagag gggtccaggc cgggcgcggt ggttcgcatc 35640

agggcgccca ttaagagggg tccaggctgg gcacggtggt tcccatcagg gcgcccatta 35700

agaggggtcc aggctgggca cggtggttcc catcagggca cccattaaga ggggtccagg 35760

ccgggcgcgg tcgttcgcat tagagcaccc attaagaggg gtccaggctg ggcacggtgg 35820

ttcccatcag ggcgcccatt aagaggggtc caggccgggc gcggtggttc gcatcagggc 35880

gcccattaag aggggtccag gctgggcacg gtggttccca tcagggcgcc cattaagagg 35940

ggtccaggct gggcacggtg gttcccatca gggcacccat taagaggggt ccaggccggg 36000

cgcggtcgtt cgcattagag cacccattaa gaggggtcca ggctgggcac ggtggttccc 36060

atcagggcgc ccattaagag gggtccaggc tgggcacggt ggttcgcatc agagcacccg 36120

ttaagagggg tccaggctgg gcacggtagt tcgcatcgga gcaccagtta agagggatgc 36180

aggccgggca cggtggctct aatccgagca ctttgggagg atcacccgag gtgaggagtt 36240

cgagaacagc ctggccaacg tggtgaaacc ccgtctttac ggaaaaatac aaaatttatc 36300

agggcgtggt ggcgggcgcc tgtaatccca gctattcggg aggctgaggt gggagaatcg 36360

gttgaacgcg ggaggcggag gttgcagtga gccagtgagc cgagattgtg ccgctgcact 36420

tccagcctgg gcggcagagt gggactccgt cttggggaaa aaaagggtag tccaggccgg 36480

gcgtggtggc tcaggcctgc aattccagca ctggaggagg ccgcggcagg aggatcgctt 36540

gagaccagga gttaagagac ctgcctgggc aatatagtga gacccctgtg tttgtttgtt 36600

tgttgagacc cttgtgttaa agcaaactaa atatggcctg agaaggactc cgtaattcta 36660

tatttgagtc cttgtggatg aactgcaacc taacttaata ggtacaaaag attgaaaacc 36720

taagttaggc cctgcgtggt ggctcacgcc tgtaatccca gcactttggg aggccgaggc 36780

gcgcgaatca cctgaggtcg ggactttgag actagcctga ccaacatgga gaaaccccgt 36840

ctcttctaaa aatacgaaat tagccagacg tggtgactca tgcctgtaat cccagctact 36900

cgggaggccg aggcgggagg atagcttgaa cccggaaggc ggaggttgca atgagccgag 36960

attgcgccat tgcacactct agcctggtca acaagagcgg gaaactccat atcaaaaaaa 37020

aaaaaaagga agcctaactt acgttttatg cgcctgtaac cgccactgag tgttggccaa 37080

tcccagcagc ccagcagctg tacttcatcc actcacaggc tgctgagcat tcaaactgtg 37140

ttcaaataag gcaaacgctg agcggtaacc aatccagctc tttctgtacc tcacttccga 37200

tttctgtatg tcactttcct tttcttgtcc ataaatttgt tctgaccacg aggcacccct 37260

ggagtccgtc tgaatctgcc gtgactttcg gagctgcccg atttgcaaga ggttcattgt 37320

tcagttgaac tcctttacat ttatttattt tatgtatttt attttattta ttttatgtat 37380

tttattttat tttatgtatt ttgttatttt attttatttt atttatttta tttttatttt 37440

gtttcatttt tatttttatt ttattttatt tttattttca tttttttgag acggagtctc 37500

gctctgtggc ccaggcggga gtgcaatggc accatctctg cgcactgcaa cctccgcctc 37560

ctaattcaag tgattctccc gcctcagcct tacgagtagc tggtattaca ggtgcccacc 37620

accacacctg gttaattttt tctgttttta gtagagatgg ggattcacca cgttggcccg 37680

gctggtcttg aactcctgac ctcaagtgat ctgcccacct cagcctccca aagtgctggg 37740

attacaggtg tgagccactg tacctggccc cctttaaatt taattcggct gaagtttttc 37800

ttttaacaga tggtgtcaga agcggagtcc agagtagagc ttctaggaac cttcgggagt 37860

gctgagtgaa cacgcaaggt gcctgcggaa cccacttgtg tccattgacc tctccgagtg 37920

gccggggatc gtgggtaagt tccctctcgg atttcagagc tccacagatt tgtgttttga 37980

gctctccgag tttcattgag caaatatctg atccaaactg ggtttggaag ctgtgacaga 38040

aactggactg gttctgggaa tggatctgat gtggtaatta gctggcttgg acccagttag 38100

aggcctctta catgtgactg ggtcagaaag aaactggtag caaatggtaa tattgcagga 38160

ggtgtaaaat ttggctttta aaaattaaca gggatgtttg tgttctaccc ctttgtttca 38220

tttttcttgt gcacttaggt attaaaaaaa aaattactgg gtaagttaat caagggaacc 38280

tgagagtaat gaatgccaat attttaggta aaaatgggat cttttttttt ttttttttga 38340

gatgaagtct cgtttggtcg cccaggctag agtgcagtgg cgcgatctcg gctcactgca 38400

agctccacct cccaggttca cgccattcta ctgcctcagc ctccccagta gctgggacta 38460

caggcgcccg ccaccacgcc cggctaattt tttttttttg tattttcagt agagacgggg 38520

tttcactgtg atggccagga tggtcttgat ctcctgacct cgtgatctgc ccgcctcggc 38580

ctcccaaagt gctgggatta caggcatgag ccaccacgcc ctgccaaaaa tgggatcctt 38640

aaattctgaa aaactgagtt ccttctggct tatacattag gcacgggaag gagcaaagac 38700

ttacagaaat ggcaaaatct gcctgagcgt ggtggctcac gcctgtaatc ccagcacttt 38760

gggaggctaa ggtgggtgga tcacctgagc tcaggagttc gaaaccagcc tggccaacat 38820

ggtgaaaccc atctctacta aaaagacaaa aattagtggg gcgtggtagc ttgtgcctgt 38880

agtcccagct acttatgagg ctgaggcacg agaattgctt gaacccagga ggcggagttt 38940

gcagtgagtg gagattgcac cactgtactc caacctgggt gacagagcga gactctgtct 39000

caaaaaataa aaatggctgg gcgcagtggc tcacgcctat aatcccagca ctttagaagg 39060

ccaaggcagg tggatcacct gaggtcggga gtttgagacc agcctggcca acatggtgaa 39120

actctgtctc tactaaaaat acaaaaaatt agccaggtgt ggtggcatgc acctgtaatc 39180

ccagctactt gggaggctga ggcgggagaa tcttgaaccc aggaggctga ggttgcagtg 39240

agccaaggtt gctccattgc actccagcct gggcaacagg agcgaaacac tgtctcgaaa 39300

ataaataaat aaataaataa aaattaaaaa attaaaatga gggctcccga aagttaaatc 39360

tgctaatctt tcagcttcgt tactatcctg atccaaagga agcagactgc agcaccagtt 39420

ggctgacttt ggataagtaa tggggtccat tttacctgag taaagtatgg gatggggtca 39480

gaggccctcc cctcagtaaa gtcccccttg gttaaaaatg ggttaaagat gacagggccc 39540

agctggggtc aagtttgagc cttgcgagtt cagtattggg tgctaagcac agtggccagt 39600

gtctgtgttt tgtcacgtat tttgctttgg cgccatgatg aaaaaatgtt aattggctta 39660

ccccacgcaa ccccttcggc cacacttgca aaactaagag gcttttgctt aaggttccat 39720

aaaacagaaa aagaatttcc tttgtgatgc cgcaggcttg gccctcagga ctatagtgtg 39780

gcaagcagag tcactaggac cacctgggca aagggaacct agaagcctgg catgctggca 39840

aaaggtaaag tatttcttac caattagact gtggcccctc tgtgcaaact ggttagatga 39900

acggtaaaaa tcactgttta tctcctctgt aatgctttga ttaatacaaa aaagaattct 39960

gaggttggtc ttaggaagct gtaataaatc tggtatgctt tgtgtctttc tgtattctgt 40020

cacgaagagg ggtaccttcg gatgaaatgc gtgcccaggg ccgcataagc ccgctgttca 40080

agacggccca gcaaactggt caatcatgtc cttggaagct taacctccta ataccatgtg 40140

gccctgcttt ctcttttcac aacggcaggc caggttcagg gttccattcc cggcttactc 40200

agcgagtact ttctggtgtc acctttacca tgtgttgatt ctcttccctc tgtttcttat 40260

agaacacaga aatattagct gtttggccta gccaaggttg ggtaataaaa gatttaaaag 40320

gactttttaa aacaagcgct atagttaaaa ctcaggtaca ttaaaagtgg atatttggcc 40380

aggcatcgtg gctcatgctt gtaatcccag tacttgggga ggctgaggca ggtggatcac 40440

ttgaggtcag gaattcaaga caagcctggg caacatggca taactctatc tctactaaaa 40500

aaaaaaaatt agctgggcat gttggtgcac gcctgtaatc ccagctattc aggaaggcgg 40560

gaaaattgct tgaacccggc aggcggagct tgcagtgagc caagatttca ccactgcgct 40620

ccggcctggg tgacagagcg agactccgtc tcaaaaaata aaaataaaat aaaaataaaa 40680

taaaataaaa taaataaaat aaaagtgtag tgctcgcttc agcagcacat atactaaaat 40740

tggaatgcta cagagaagat tagcatggcc cctgcacaag gatgacatgc aaattcatga 40800

agcgttccat attttgtgca tcctggcaag atcatttctg tatctgctaa ctagcactaa 40860

agaaatagtg tgaatctaag caaaaatgag tggcacccaa aaacaaaatt gtggttttca 40920

ttaaaaaata taaatattta tataataaaa tatatatgat atataaaaaa taaaatatgt 40980

ataataaaat atacataata tatataaaat aaaaaagtat ataaaaaata tatatgtgtg 41040

tgtatattca aattcttttt tttttttttg agacggagtt tcactcttgt cacccaggct 41100

ggagtccaat ggcacgatct cggcttaccg caacctccac ctcctgggtt caagtgactc 41160

tcctgcctca gcctcccgag tagttgggat tacaggcacc tgccaccacg cccagctaat 41220

ttttgtatcc ttagtagaga cagggtctcg ccatgtaggc caggctggtc tgaactcctg 41280

accttgggtg atccacccgc ctcggcctcc caaagtgctg ggattacagg tatgagccac 41340

cgtgcccggc aatgtatata ttcaaattct aacagcctgg gactccttgg gaaaaacagg 41400

aggcaccaga gacgccattt tggaaaaaaa cctgttttcc tcatggaacc acaggaattg 41460

gaaatggata gattcccctt cagaaatcta aggctctgtt ctttttggga ttcaggatct 41520

ggtataaaaa tgggaccctt aatttttgga gacctgtgtt gctttctgct gtgcccactt 41580

attatattgg gccctagaaa ctgcatgctt tcctggtcct ttttgtccaa ggactccacc 41640

ctaaagccag taatctcaaa taaataaata aacacctaaa tcaaatattt tgaaagaaca 41700

ataaaaacta atgcctttta gttcacataa ctttagtaat ctttgggaaa taaaaagttt 41760

taaagattat tgggaaaaat gaagacattt agtctaaatt aggcaggtca gatattaggt 41820

ttgttcgatg ctttaaggtc atcaactgct tcttaggctt ttgaaaattg ttcagtttac 41880

ctacttggga gcattagatt ctagataagg cctgggaaca tgtggaatta gccatgcact 41940

ctatctatgc aaaggttata aagaaaagag attttttttt taattaacaa aaaccacagg 42000

tggttgtgct ccgcctaccc agcacagcag gtgagatggt gcaccctcag tggactgaag 42060

aggccatggg atcccacctc agccggatag cacgctggcc ttcaccttca ttgtgccatg 42120

gcagctttcg tgtgagccac tgactcgtgt acatgttaga ctccttggtc tataagaaag 42180

catcttgtat ggtaaattgt tgttctaaag taaaataact ggtttgttca aaaaggggat 42240

gctaaggaga agtcagaaag tcaaagcatg ttgtagatgg tctgggaaag tcgtgaaagg 42300

gttcgtgaaa ggaaatttaa gccaccaaaa gtaaaagttg ctagttacca ttataacata 42360

tgattgaaac tactgaaaaa atagttttac atggaaagtg tgtgagaaga gtgaaacatg 42420

tttttggtaa aatattttta aaaggcagga gaatgtcaaa tttcacctag ttgagagggt 42480

tatgtgattt ttaaattaga taagaataag ctcacggttt gaacaagttg tggaaggttt 42540

gtaaaaacta atcttgcaaa aaaaattctg tgtgcaaaca cgttgactaa atttaaaggg 42600

gtattttcca gttttccata aattaaacat tgaaatgaaa ggacaaacag ggtttctttt 42660

tttgtttttt tgttgcccag gctatgggca actatgtagc taggcatgtt ggtgcatgcc 42720

tgtaatccca gctactcagg aagctgaggc aggaaaattg cttgaaccca agaggcagag 42780

gttgcagtga gccaagatcg caccactgca ttccagcctg ggtgacagag caagacccta 42840

tctcgaaaaa aaagaaaaaa gaaaagcaat agctccctgg gtgtcaggta tcagtgagaa 42900

tgggaagtca ggtcttctct gactcctagg acattcctct ctctgtcttt gggggttgaa 42960

ttccttgtta gaactccatc ttgttaaagg ggcctcgcac ctgcccttgc cttctgcagc 43020

agccctcctt aatcctctcc aaacttccat ctccagacca ccggattctc ctcatgagtg 43080

ccctgggtgg acattggtga aaggtcactg ccttcaggac acaccatagg tacagtcacc 43140

gaggtaaccc agttcaaaga agagcaggat gttcagtatt tacaaaggga tctagcaatt 43200

ggtccaaacc agtcagtgct ggcatgcacc gttcatcgtg ccatcagtgg caagatggag 43260

atctggcaat gcatggctca cagtaagata cacgccctgt gccattccac cctatcatta 43320

gaaaaggagt cttctttttc tttctttctt tctttttttt ttttttttgg agacagggtc 43380

tcactctgtt gcccaggttg tagtgtaatg gtgcaatctt ggctcactgc agcctctgcc 43440

tcctggggtc aagtgatcct ctcacctcag cctcctgagt ggctgagatc acagatgcca 43500

gccaccacgc ctggctaata tttttgtttg tttgtgttat gttttttgag actgagtctt 43560

gctctgttgc acaggctgga gtgcaatggc gcaatctcgg ctcactgcaa cctccacctc 43620

cctggtacaa gcaattctcc tgtctcagcc tctcaagtag ctgggattac aggcgcctgc 43680

cactacgccc gcctaatttt tgtattttta gtagagacga ggttttgcca tgttggccag 43740

gaatggccag acctcaaatg atctgcccgc ctaggcttcc caaagcgctg ggattacagg 43800

tgtgagccac tgcacccagc caatttttgt attttttttt tttttttttt ttgtagagac 43860

agggttttgc cgtgttgccc aggctgatct cgaactcctg acctcaggtg atcaccttcc 43920

ttggcctccc aaagtgttgg gattacaggt gggagccacc atgtccagcc gctttttctt 43980

tttaaaaaaa attttttttg acataacaga caacaccact ctcttttatt tttttataga 44040

gacggggtct cattatgttg accagtctgg tctttttaaa ttttattttt ttgagatgga 44100

gtctcactct gttgcccagg ctggagtgta gtggcgggat cttggctcac tgcaacgtct 44160

gtaccccaga ttcaagtgag tctcctgcct caggcttcca agtagctggg attacaggct 44220

catgccacat cgctggctaa tttttttttt ttgtatgttt ttaaatttta tcttttaagt 44280

agagacaggg tttccccatg ttggccaggc tggtcctgaa ctcctgactt caggtgatcc 44340

accctcctca acctcccaaa gtgctgggat tacaggcatg agccaccata cccagccttc 44400

ctgctatttt tgaactacct gtaccaggct ggtttctctg ggtgagggga gtggtcaggc 44460

agaggcaggc cccgtcatgg taagcagcta ggagactgct agagattgac agggcactca 44520

tgagtgccct gggtggacac tggtgaaagg tcactgcctt caggacacac cataggtaca 44580

gtcaccaagg taacccagtt caaagtagag caggatgttg ggtatttaca aaggagatct 44640

agcaattggt ccaaaccagt cagtgctggc atgtaccgtt catcgtgcca tcagtggcaa 44700

gatggagatc tggcaatgca tgggctcaca gtaagataca cgccctgtgc cattccgtag 44760

acgtagatca agcgattatt actgcagaaa tctccgtgga cgtagagcaa gcaattatta 44820

ctgcagaaat tgcaggagca ggttctgcag catgtcacat ccaaagaagc ttcccttgca 44880

agttggtaac cttgagggct tcaataccac acagagttgg tctaggtgat ggctgaccat 44940

ccaggcataa taatttgggg aacaattcca ggatggaggt tatgttgcta gacttgtttg 45000

tccgggaaag tggtagacag gaagttcctg tctgcacacg tctcctggct ccagcccttt 45060

cccctgtcca tctgtcctac agactccgcc tcaggcatgt tgtcttggag gacgtgcaga 45120

aaggcagcca tgcactttct aggtttcgat gccgtcttag agtcactggt caccaggtct 45180

gggtcaaacg ggagtctctg gctgtcaaga ctggactggg aggaatcaca tcgcttcttc 45240

accatcagct ttatctgctt tgccttttcc gacttcagct ttctctgcag ggggtatcag 45300

tggggttgac aaagattgac atctttttgt tatcctcatc cctaatcttg ccactgacat 45360

tcttcagtgc ctaggcgatg ctggcattct ccacaaagat gtgggcctgc attttctcat 45420

agcgaaactg aatcaggagg aaggggcatt gcattgactc tgaatcaaat tcagcagcca 45480

cttctcattt tatgcagatc ttgaactagc agctccctaa ggtccgatct ggcctgtttc 45540

cctctgttct tttctctgga ggtttttggc gctttctctg tgttaacatg ggtttggtct 45600

tgtttatggc aagtgcctct ccatggctgg gtgcggtggc tcacgcctgt aatcgcagca 45660

ctttgggagg ccaaggctgg cgaatcacca gaggtcagtt cgagaccagc ctagccaaca 45720

tggcgaaacc ccgtctctac taaaaatata aaaaaattag ccaggtatgg tggcgggcgc 45780

ctgtggtccc agctactcag gaggctgagg caggagaatc gcttgaaccc gggaggcgga 45840

ggttgcagtg agctgaaatt gtgccattgc actccagctt gggtgacaga gtgagactct 45900

gtttaaaaaa aaaaaaagta agtaaataaa taaatgttga tattatgtag ccatgttatg 45960

tagttttgca tagcctttct ctgtcttata gctatgaaag acaatttcca aaacatagcc 46020

tagaatggag aattgaagat aaagtgagaa ctgttgggca tagtaaccta aaaagggatc 46080

ccctctggtg tggaaagatt gttccaacgc aaaagggact gagatccaga gatggcagtt 46140

gtttgagatc tggaatcttc agaagaggta gatcagactt gctcaggaaa taatgggtcc 46200

taccatcaca gcgggtacgg atggggctac aagatgagag aggctggata ggagagtatt 46260

taaggaaaaa aaaaagacag aaaaggacca aaggacagtc tgtgagaccc cagagaatgc 46320

attaaacctg aagaatttct gaacaatgac gtctgatatt caggcttccg ttgtttcttt 46380

ttgtgctgta atcttccgtg ttttctctgc acatagctca gtaaagctct attcaccagt 46440

taacctagag tctgtcttga gataagcagg gagggctgtg acggctgcac aagggcggag 46500

tggtttgtag acctgaaaag gaacttgtaa gagctggcag atcttaacca ggagagggag 46560

cccctttcca cagcctagaa gcttctgaag tcatttaaat ggaactgaat tatatgaagt 46620

caaatgtcag ttggggaaaa agtcaaactt tattatttat ttatttattt gtaattgtgc 46680

gtatacatat atatatatat atatatatat atatatatat atatatatat aatttttttt 46740

aaattatgat ctccgtgata gtaccaaatc aaactttttt tggagacaga gtcttgctct 46800

gtcacccagg ctggagtgca gtggcgtgat ctcggctcac tgcaacctcc gcctcccagg 46860

ttcaagcaat tctcctgcct cagcctcctg aagagctggg attacaggcg cgtgccacca 46920

tgcccggcta atttttgtat ttttagtaga gatagggttt caccatgttg gccaggctgg 46980

tcttgaactc ctgacctcag gtgatccacc cgccttggcc tcctaaagtg ttggaattac 47040

tggcatgagc gaccacgctg gcccaaaccc tgctttttac tggagagacg aggtagccct 47100

aagttaaaaa aaaaaaaaaa aagaccctgt ttcacttctt ttggatatac ccggggtgac 47160

attgctagat cattctgcaa ttcctccttt tttttttttt tttttttgct gggggtggga 47220

tagagtctca ctcccaggct ggagtgcaat ggtgtgatct tggctcactg caacctccgc 47280

ctcctgggct caaacgactc tcctgcagtg aggagctggg attacaggtg cctgccacca 47340

caacctgcta attttttttt tattttttgt atttttagta cagacggggt ttcactatgt 47400

tggccaggct ggtcttgaac tcctgacccc gtgatccgcc cgccacagcc tctcaaagtg 47460

ctgggatgac aggcatgagc ctccgcgccc agtagatcat tctgtaattc tgtgtttaac 47520

tttttccccc ctaaacttaa ctgaagtgat atttaacttt ctgaggaatc accaaatcaa 47580

aggtgacttt tgagacaaat acaggaagtg gaggtggttt tcctcatgta gatgaagaca 47640

ttctaaaccc agaactgaaa acacacatgg aatcctttta agtatcagga tgccttggtg 47700

gacaaaatgg atacatccaa cttctcaagg agcttaatat tcagtattaa agctgaggtg 47760

agctcaggtg attcacgcct gtaatcccaa cattttggga ggccaaggca ggcggatcac 47820

ttgagcccag gagtttaaga ccagcctggg caacatagtg agacccctcc cccctgcccc 47880

cgtctctaca aaaaataaaa aaattacaga cgcatggtgt agtcccagct acttggcagt 47940

caaaggtaga aggactgttt gaacccagga gtgggttgaa gctgcagtga gccttgattg 48000

taccagtaaa ctttagcttg ggaggcagtt gagaccctgt ctcaaaaaaa aaaagaaagg 48060

aagaatttct ctctcccaac ttcccttggt ttctcctcgc cccccggcta aaaagtgatt 48120

ggggaccttt taaaaaactt cctgctctcc acagaatggg attacagccc cgagatgatc 48180

cataaaagat tttcaggaag aatgaagagt ttagggagtg aaggttctaa aacagtaaat 48240

agagtgtcac aaagaaatcc attcatctct ttgaaggctg ggtatttgga aaaagagaga 48300

aagaaagctt cattcagaga gcgctcgcct gtatttaggg gtgtttggag ggggcggggt 48360

gactatagaa ttagggtaaa gttcagtgtc ttgcaaatca gccctgaaat agccctgatc 48420

cctgcaggcc tgtctggttt cccaaaccct ccactagctt tcaaatttgc attaaaaagc 48480

ttttggcttg ttcattctgt tctttctttc ttttctttct tttatttatt tatttttttt 48540

gagacggagt ctggcactgt cacccagact ggagtgcaat ggcgcgatct ccgctcactg 48600

cagcctctac ctcccgggtt caagcaattc tcctgcctca gcctcccgag tagctgggat 48660

tacaggcgcc caccgccaca cctggcttat ttttttaatt tttaatagag atggggtttc 48720

tccatgttgg ccaagctgtt ctcaaacttc tgacctggtg atccacctgc ctcagcctcc 48780

caaagtgctg ggattacagg cgtgagccac tgcgccaggc cctcattctg ttacttcttt 48840

ctttcttttt tcattaattt aagttctggg atacacatgc gggatgtgca ggtttgctac 48900

ataggtaaat gcgtgccgtg gtggtttgct gaacctattg acctaggtat taagcctcac 48960

agccattagc tatttgtcct gatgcccttc tgcctcctgc ccattctttg ttatttcttc 49020

atattccaat taatgttcat tttgaggacc tggctctggc caggcaaggt tccagtctgg 49080

agtcaaactg aggaaagtat tacctttcac aaacacttag accacgttgg gaggggagaa 49140

tgattgtcct ccacatgcgt atgggtgtgt tataaacccc aggaaggtaa atactaagca 49200

aaggcgtgca cagataatta ttgcaactga tgattccacg tcaattgtac tgtactgtga 49260

gtaggtttat aggctgccgt gacaggggcc atcacctggg tacgagggtg ctggcttcct 49320

cttgaagacg tgactttaaa attgtgagtt gaatagatgc aaggatattg gcccagtgga 49380

ggaactttga ggggaggggg cagggtatcc tggttgggag gaaattacat acagaaattg 49440

gggagtggtg acaacattga ctcttccagt aattgtctgg gtgtggtaac tcaggactgt 49500

aatcccagca ctttgggagg ctgagatggg aggattgctt gagcttggga gttccagatc 49560

agcctaggta acacgaggag atctaaaatt gaaaaaaaac gtagctgggt gtggtggctc 49620

attcctataa tcccaacaac tttgactcag gaggctgaag tgggaggatc ccttaaggtc 49680

aggaggttga ggctgaagtg agctagcact ttgggaggcc gaggtatgtg gattacttga 49740

ggccaggagt tcgagaccag gctggctaac atggtgaaac cccatctcta ctaaaaataa 49800

aaaaattagc caggcatggt ggtgcgcgcc tgtaatccca gtgactcggg aggctgaggc 49860

aggtgaatca cttgacccct ggaggcagag gctgcagtga gccaagatca taccactgca 49920

ctctagcctg gacaaccaga gtgaaactct gtctcaaaaa aacaataata attttaaaaa 49980

aattttttaa ataaaaaaat taacaaatga cccgcacatc tggggtgaag taagacgagg 50040

ctggcagcat agctctgtca cctgtggtta ggattaatta agctcagctt taaaggctgg 50100

atcttggcct cctgggtttt aattttgact tctccacttc tcagccatgt tttgttttgt 50160

tttgtttgct tgaggaggga caaatcaggc cagcattgag cctctttccc ccgtgtgtca 50220

actgagattg ataattacaa gattaaatga tgcttgtctg tgattttagc atctgttcag 50280

cccatcattc atttcaagag tattgactgg gcttcagggt aggttccatg ttagtactgg 50340

ggacgccgtg ggtgggaggg agactgggtc tccaccctcc tagagttata gtctaggcct 50400

gcggttaagc agatttttgt gcaggaatca aacccactac actacctgtc agaaatcaag 50460

ccctgttcca ggcactggga aactcagaat ttaaagggga tgtaccaggt ctctgtcctt 50520

gcggagcatt atcgatcagt agaagagaga aaataaacaa gcaaacacct gagacaattg 50580

taatagcaat aatgtgatat caataaaatg aaacagagca acagaaatag ggtagatggg 50640

ccgggcgcgg tggctcaccc ctgtaatccc agcactttgg gaggccgagg tgggcggatc 50700

acgaggtcag gagatcgaga ccatcctggc taacacggtg aaaccccgtc tctactaaaa 50760

atacaaaaca ttagccgggc gtggtggcgg gcgcccgtag tcccagctac tcgggaggct 50820

gaggcaggag aatggcgtga agccgggagg cggagcttgc agtgagccga gatggcgcca 50880

ctgcactcaa gcctgggcaa cagagtgaga ctgtctcaac aaaaaaagaa aagaaaagaa 50940

acagagtaga tggtcttaga aatgacaagg aatcccaatg agaggttcta cctgggacaa 51000

aatagaaaaa tgggaaatca taataagctc tggaattggg gcagtggtat tgtttatttc 51060

ctagttgtgg caaatctaat gtaaaatatg gacattaaag aaaactgggt ggagaacata 51120

gagaactttc tgtacttttt gcaacttttc tgtaagtcta aatttattcg aaggccgggc 51180

gcagaggctc ctgcctgtaa tcccaaaacg ttaggaggct gaagcaggta gatcacttga 51240

gcttggtagt ttgagaccag cctgggcaaa aaagtgagac ttcatctcca ttaaaaattt 51300

aaaaaattag ctggctttgc tggtgagcac ctgcaccaga ctggttagcc tgggcagcat 51360

agtgagaccc cgtttctata aataagataa aaatcagctg ggtgccaggc gtgatggctc 51420

atgcctgtaa tcccagtgct tcgggaggcc gaggggggcg gatcacctga ggtcgagagt 51480

ttgagaccag cctgaccaac atggagaaac cccgtctcta ctaaaagtac aaaaattagc 51540

tgggcgtggt ggcacgcgcc tgtaattcca ggtactaggg aggctgaggc aggagaatca 51600

cttgaacctg ggaggcggag gttgcagtga gctgagatca ctccactgcc ccccagcatg 51660

gctacagagc aagactccgt agcgggatat tgcatgcatg taatcccagc tactctggag 51720

actgaggcag gagaatcatt tgaacctggg gggcggaggt cacagtgagc caagatcatg 51780

ctattccagc ctgggtgaca gagtgagact acctctcaaa aaaaaaaaaa agaaaaagaa 51840

aaagaaaaag aaatgcattt acagaagcta ggaaaatcaa attcagggag gaggctggag 51900

ggcaaagagt gggtgtggcc agactttaat ccattctcca tctgtggggt ggtccctagg 51960

gtataaaaga gctccagggc ccctgccctg cctttcatcc tcaggatggg aagtcctcac 52020

agagttcagt aagtattggc ttcccatggt gtttctgcac ttcccccggg gtcatatgag 52080

cctcacattt aggttgtaag actcccccta aaatctccaa cactgagtcc tctctatttg 52140

tagtcatgac tccgacacct tgattctctg cccctaatac tctttttttt tttttgagac 52200

ggagtttcac tcttgttgcc caggctgcag tgcagtggtg tgatcttggc tcactgcaac 52260

ctctgcctcc cggattcaag caattctcct gcctcagcct cccgagtagc tgggattaca 52320

ggcatgcacc accatgccca gttgattgtt tttgtatttt tagtagaggt ggggtttcac 52380

catgttggcc aggtctcaga ctccaagcct cgagtcatcc gccctcttca gcctcccaaa 52440

gtgctgggtt tacaggtgtg agccaccatt cctggaccgc attattgatt ttttaaaatt 52500

tttggatttt gttcaattag tgattgagtc tcactgtgtt gcccagatgg gcttgtctcc 52560

caggctggag tgcagtggtg agatcgtgac tcacagtagc ctcaacctcc ctcaccccag 52620

cgtcccgtgt agttgggatg gcaggcatac aggaccatgc ttagctaatt tttttttttt 52680

tttgtagggt tggggtctcc caccattgtc taggctggtc atcaacacct gggcacaaac 52740

atcctctcac cccggcctct gagccactgc acccagtagg ttttaggttc attttatttt 52800

ttatttaggt gtccacaaag gcagcttttt ccttttcttt attttttaaa gagacagggt 52860

ctcactctgt tgcccagggt ggagtgcagt ggccagatct tggctctctg caacctctgc 52920

cttccagact caagtgatcc accttcgtct ccagagtagc tgggactata ggtgtgcaca 52980

aacacaccca gctgagtttt aaatcttttg tagagaaggg ggcctcagta tgtttcccag 53040

gctggttttg aagtcctgtg ctcaagcaat ccttctgcct tggcttccca aactgctagg 53100

attacaggtg agccaccaca ccgggcctgt ccacgtaatt taaaaggtgc tggacgtgtg 53160

cggtggctca cgtctgtaat cccagcactt tgggaggctg aggtgggcgg atcacgaggt 53220

caggagttcg agaccagcct ggccaatatg gtgaaaccct gtctctacta aaaatacaaa 53280

aattagccag gtgtggtggc aggcgcctgt aatcccagct tctcaggagg ctgaggcagg 53340

agaattgctt gaacccagga ggcggaggct gcagtgagct gatgatcgct ccatggcact 53400

ccagcctggg caacagagca agactaagaa aaaaaaagaa agaaaaagaa aaagaaaaca 53460

aagtaatgta aaaggtgtta ttcaggctgg ggacccccaa agtgctggga ttacaggcat 53520

gagccccagg gcaggcctag gatttgcatt acaaggctct gcattgttcc tgagaactgg 53580

agagagtgct caatctacct ttcgctattg agcaacattc agaagtcagt tttaaatctc 53640

ttatcttctg catagaggat gtgcctgcag tttccaggta ctggacaata cagggagggt 53700

acttctcagt ctgtggcact cagccttgag ggcactttct ggtgccagaa tgaaagtgct 53760

gtcatagctg aggtccaatg actgaggcga gcaccgaaga aacaccatgg gggggagggg 53820

gggcgggggg ctccaggagc cactgcaggt aaaagctaaa ccgtggtgct tgttctgcgc 53880

cataaactgg acttcagccc agctttaagg aaatgaccac ggtctctggg ttgtgaggct 53940

gggttccgta gcagctataa tgcttggatt cagggatagc ctgggtcaca ccattgccag 54000

agaagaggca accaatccag accccaaaag cgggttcttg gatctcttgc tggaaagaat 54060

ttcgaggcaa gtcacaacac agcgaaagaa gcaagctgag gcctgggctc aatcctcctg 54120

cttcgtcctc ccaaagtgct gggattacac gcatgatccc ctgtgccctg ccaatatttg 54180

gtaattataa ctggtggtca gcttacaatg tggctatttc cagaccataa gtaccaactc 54240

tataggtgcc ttgtgagtga gtgccttgct acttcaagca gttactttcg tgtgttatta 54300

aaccagaggc ctgctaagcc tctgtgccta gtaaagatta caatttggaa tgtcagcccc 54360

cagctgtcct gtgtgattgg caggttaggt ccttgttgaa ccagaagcaa ctggccagtc 54420

gtacagatgc aactttcgga ccagtagaaa atttctatgt caacctggag acatagggat 54480

tgtacctttc tgatacccta ttagtaaata aatagtattt atttataaat tatgtataat 54540

agtatttatc tgaatatatc ccaagcagcc caagtgtgtt ccagacataa catttttatt 54600

tacatttaaa tatcactaag attagagata cacatctaac tcaggttttc aactagtctt 54660

accattgaaa gaactattgt ggcaggacgc agtggcgcac gcctattatc ctagcacttt 54720

gggaggccga ggagggcgga tcacaaggcc aggagttcga gaccagcctg gccagcatgg 54780

tgaaacccca tctctactaa aaatacaaaa aaattagctg ggcatgatgg cacaggcttg 54840

cagtcccagc tactcggtag gctgaggcag gagaactgct tgaacccggc agacggaggt 54900

tgcagtgagc cgagatcgcg ccaccgaact ccagtctggg caacagagca agacgcttgt 54960

cttaaaaaag agagagaaag aactacttgt tctctgtaag ccatgctggt ttagccaaaa 55020

tgctgcagtt gtgtatggcc agggacgttg aggctgcagt gagccgacgg ctgtgttgcc 55080

gctctgcagt ctgggggaca gagcaagaac ctgtctcaaa aaaacaagaa gttagtgaga 55140

cagacatgcc gttctgttac ttttcccctt gctggttcaa agctaggtag ttgccaatta 55200

aatgtaatct tgtagagcaa aaatgttttt cattttttga gacaggatct cactctattg 55260

cccagcctgg agtgcagttg tgggaactca cagcaacctc cacctcctgg gtttaggcag 55320

ttcttctgca tcagtctccc gagtagctga gactacaccg gtgcccagag ggcagtggcc 55380

agatcttggc tctctgcaac ctctgccttc cagattcaag tgagccacct ccgcctcctg 55440

agtagctggg actataagta cgtaccacca tacccagtta atttttgcat tttttgtaga 55500

taagggggct aagtatgttt cccaggctgg tttcgaagtc ctgtgctcaa gcaatccttc 55560

tgccttggct tcccaaactg ctaggattac aggtgagcca ccacaccggg cctgtccacg 55620

taatttaaaa ggtgctggac gtgtgcggtg gctcacgtct gtaatcccag cactttggga 55680

ggctgaggtg ggcggatcat gaggtcggga gttcgagacc agcctggcca atatggtgaa 55740

accctgtctc tactaaaaat acaaaaatta gccaggtgtg gtggcaggcg cctgtaatcc 55800

cagtttctca ggaggctgag gcaggagaat tgcttgaacc caggaggcgg aggctgcagt 55860

gagctgatga tcgcgccatg gcactccagc ctgggcaaca gagcaagact aagaaaaaaa 55920

aaagagaaaa agaaaaagaa aacaaagtaa tttaaaaggt gttattcagg ctggggatcc 55980

ccaaagtgct gggattacag gcatgagccc cagggcaggc ctaggatttg cattacaagg 56040

ctctgcattg ttcctgagaa ctggagagaa tgctcaatct acctttcact attgagcaac 56100

attcagaagt cagttttaaa tctcttatct tctgcataga ggatgtgcct gcagtttcca 56160

ggtactggac aatacaggga gggtacttct cagtctgtgg cactcagcct tgagggcact 56220

ttctggtgcc agaatgaaag tgctgtcata gctgaggtcc aatgactgag gcgagcaccg 56280

aaaaaacacc atggggggga gggggggcgg ggggctccag gagccactgc aggtaaaagc 56340

taaaccgtgg tgcttgttct gcgccataaa ctggacttca gcccagcttt aaggaaatga 56400

ccagggtctc tgggttgtga ggctgggttc cgtagcagct ataatgcttg gattcaggga 56460

tagcctgggt cacaccattg ccagagaaga ggcaaccaat ccagacccca aaagcgggtt 56520

cttggatctc ttgctggaaa gaatttcgag gcaagtcaca acacaatgaa agaagcaagc 56580

tgaggccggg cgcggtggct cacgcctgta atcccagcac tttgggaggc tgaggtggat 56640

ggatcacctg agatcactat ttttcttttt attttctgag atggagtttc actcttgttg 56700

cccaggctag agtgcaatgg cacgatctcg gctcacagca acctctgcct cccgggttca 56760

agccattctc ctgcctcagc ctccggagta gctgggatta ctggcatgcg ccaccacgcc 56820

cagctaattt tgtattttta gtagagacgg ggtttctcca tgttggtcag actggtctcc 56880

aactcccaac ctcaggtgat ccgcccgtct cggcctcaca aagtgctagg attacaggcg 56940

tgagccaccg cacccagcct gttttctgca tttttagtag agacagagtt tcactatgtt 57000

ggccaggctg gtctcaaact cccgaccttg tgatctgccc gcctcggatt cccaaagtgc 57060

tgggattaca ggcgtgagcc accgcgcccg gccttatttt tctttttttt tttttgagac 57120

aggatcccac tctattgccc agcctggagt gcagtgcagt gatctcagct cactgcaacc 57180

tctgcctccc gggttcaagc aattctcctg catcagccgc ccaagtagct gagattacag 57240

ccgtgcctgg cttatgtttt gtgtttttag tggagatggg gtgccaccat gctggccagg 57300

ctggtcttaa actcctgacc tcagttgatc ctcccacctc agtctcccaa agtgctggga 57360

ttacaggtgt gagccaccgc gcccagcaag cataagattt tttgacacat ggaggatcac 57420

ttgctggatg tgcactgaga agtaatgtgc ccatttccca gccacactct cccaaaggct 57480

tttacaggcg ctgtccccgt gaggtggtag aacagaatgt cagaattcaa cacattccaa 57540

cactgtcttc tgttgagctg taggaggact gcctgtggag ggacatgggt tctatgctac 57600

cttgcaggtt ttccctgggg cttgtgatca ctgtctcctt ggtgacatgc aatctgggag 57660

tgctttctgg agtctgataa gaacatactg gcacattgcc tgtgacgtgt gggacccagg 57720

cattttcagt gctgaggtga gtccttgatt gtaactgatg gggactggta tcttaactcc 57780

aggttactga agactaggaa tttcttgcct tacagtaaaa ttgcaaagaa gagtagcata 57840

agaggtagat cagaagactg cagaatagat taacaaatga atgcccaaat gaggatgcta 57900

aacttggatt tgcatcaaga agacagatat tggtcgggcg tggtggctca cgcctgtaat 57960

cccagcactt tgggaggctg aggcgggtgg atcatcctgg ctaacacggt gaaaccccgt 58020

ctctactaaa aaaatacaaa aaaattagcc gggcgtggtg gcaggcgcct gtagtcccag 58080

ctgctgggga ggctgaggca ggagaatggc atgaacccag gaggcggagc ttgtagtgag 58140

ccgagatcgt gccactgcac ttcaccctgg gcgacagagc gagactctgt ctcaaaaaaa 58200

aaaaggacag gtattaggat agttttgaag cccacttgct cctatcatgg cattgaagca 58260

tcagtttgct gccaatcact ccttcttctg tgttcataga caggactgtg gaaatgtaca 58320

cttggattct ttcacaggta atgctgagaa atttaaaacc aaaaacggcc gggtgccgtg 58380

gctcctgcct gtaattctag cactttggga ggctgaggcg ggcggatcac aaggtcagga 58440

gttcgagagc agcctggcca atatggtgaa accccatctc cactaaaaat acaaaaatta 58500

gcctggcata gtggcgggcc cctgaagtcc cagctactca ggagactgag tagacgagga 58560

gaatggcttg aacatggggt gcagaggttg cagtgagctg agatcgcacc gctgcactcc 58620

agcctgggca acaagagtga aactctgtct caaaaaaaaa aaaaaatttg aaattaaagt 58680

gtatgattag agtgcatgat tggttctgac tggatgtgta acatgtgcat agccttctca 58740

taaatcaaag gtttttatgt gaaacctacc actagtgatg taactgaatt tgtgtaaatg 58800

ctggtgtgtc tgccttttcc tagcaactcc ctgggcagcc tgaggattct ttgaggttat 58860

tttttttcaa gatctggctg ggagcttctg catgagtgct gtgtgaacgt cctccatctg 58920

cagtgtgagg acagcgaacc ctagtgtgag ttaaccacgt aggaagagtt tgaagtcaga 58980

catgacattc agactgaggt cctcaaaact gaggggcatt ttctgtggtt tgaaaggaaa 59040

gtgcacccag ttttggggat gtcaattgtg aatcctcatc ataacccatc cccagtcccc 59100

catcgcattg caccacatgg atgcacccag tggactggcc tttgcttttc tattgtttct 59160

ttttctctcc actgcaatca gctttctcac gttgaatgca ttccattttc taagcgctct 59220

cctaggcaaa aatccctcta atgcaaagca aaaatgtctg ttatgcaaca ttgttctgag 59280

accccaggaa ttgcagttaa cttggctcct tagtatgtgc ttcctaaggc gggtggggtg 59340

gctcgcgcct gtagtcccag cactttggga ggcaggcgag gcacttgaaa ccctgtctct 59400

actaaaaata caaaaattag ctggacatgg tggcatgtgc ctgtaatccc agctacttgg 59460

gaggtgcaga caggagcatc gcttgaaccc aggaggcaga ggttgcagtg agccaagata 59520

atggcactgt actgcaacct gggtgacaga gcgagactca aaaaaaaaaa aaagtgcttc 59580

ctgtgaaggt ttcctttccc ttctgccccc cataccagtc catctattta gtagatgtcc 59640

tggtttctac ccaggatatc atggaccagg ggttctgtct tccagccttg agtacaatgg 59700

tgcgactata gctcactgca gtctcaacct cctgggctca agtgatcatc ctgcctcagc 59760

ctccaagtag ctggtattac agatgcacgt gcccccattc ccagaggttc ttaaatttat 59820

gacaaaaacc cagaagcttg gctgaagcag cttgttgggg cccacccaca gattttgttt 59880

ggtgccatag acctgggttg ggatctgggg agccattttt cttagtctca atgtaaatgt 59940

ctaagcattt tctttttttc tttctttttt tttttttgag acagggtttc gctcttgttg 60000

cctaaactgg agtgcagtgg cacaatctcg gctcactgca acctctgcct cctgggttca 60060

agtgattctc ctgcctcagc ctcccaagta gctgggatta caggtgtgca ccaccatgcc 60120

cggctaattt ttttttattt ttttattttt tttatttttt gagatggagt cttgctctgt 60180

tgcccaggct ggagtgcagt ggtgtgatct cggatcactg caagctccgc ctccttgatt 60240

catgccattc tcctgcctca gcctcccgac tagctgggac tacaggtgcc cgccaccacg 60300

cccagctaat ttttttgtat ttttagtaga gacggggttt cactgtgtta gccagggtgg 60360

tctcggtctc ctgatctcgt gatctgcctg cctcggcctc ccaaagtgct gggattacag 60420

gcttgagtca ctgcccctgg ccaatttttt gtatttttag tagaaatgga gtttcaccat 60480

gtttcctgcc tatctctcta tgtatttaga cagggtctca ctatgttgcc caggttggtc 60540

tcaaactcct ggactcaagc aatcctcctg cctccacctt ccaaagtgct gggactacag 60600

gcttgagcca tcctgcctga ccatttgaga gttttttttt tttgagatgg agtctctctc 60660

tgtcacccat gctgtagtgc agtagtgtga tcttgcctca ctgcgacctc tgcctcccgg 60720

gttcaagcaa ttctcctgcc tcagcatccc cagtagctgg gattacaggc gcccgccact 60780

acacccagct aatttttgta tttttagtag agacagggtt tcaccatgtt ggccaggctg 60840

gtctcgaact cctgacctca ggtggtctgc ccacctccgc ctcgcaaagt gctgggataa 60900

gaggcgtgag ccaccgtgcc tgtcttcaac aatgatttct aattgcattt tatggtggtg 60960

attgaacata atgtactttg gtccttctcc attgtatgcc tcgtgtatta tgtgtctcat 61020

aacgtctgtc ccgtcctatg ctatgtgtgc atttgaatgt tggggagggt tcagtagatg 61080

tccattgagt tggtttgcaa acttatttga aacttctact tacttctgcc taattctgct 61140

tattattgag agtggactgc tctgggatgc cttttgtttg cactctgatt tcagatggac 61200

cctgagaatg ctcaagcccc aaaccctcct tgggaagtga agctcaggct gtgatttcaa 61260

gccagggggc gtttttctat aactggatga aaagcacctc cagagcttga agctcacagt 61320

ttgagagcaa tcgtctaagg aagttgatgg caatgttaat agttttttaa acaagcatga 61380

aatcagattc ctgtgtttac ttctggatgc tgttgatcca ggaaatgtac ttagaaaatt 61440

cattttaacg gaaatagtca taagcgtctt ggtaatttca tgaagctaca gtgagtaaat 61500

tgcctgagaa tttccctgcc tgaaaggtct tcaggagtgt aatttttttt taatatagct 61560

tatttaatac aaatagagat gcggtctcac tcttgcctag actggtgtgc agtgtcatgt 61620

tcacagctca ctgcagcctc tcaacctccg gggctcaagc atcctcctgc ctcagcctcc 61680

caaagtgctg gaattacagg tgtcagccac cactccctac tcatacttgt atatttctct 61740

ttaaattctg ttgaggtttt atggcctagc ctgtggtcta tcctggagaa tatttgtgtg 61800

catgacaatg tgtatatata tatatatata tatatttttt tttttttttc ttttttttga 61860

gatggagttt cactcttgtt gccctagctg gagtacagtg gcatgaatct cagctcactg 61920

caacctccgc ctcccgggtt caagcgattt tcctgcctca gcctcccgag tagccgggat 61980

taccctccac cacacccggc taatttttgt atttttagta gaggtggggt ttcgccatat 62040

tgaccaaggt ggtcttgaac tcctgacctc aggagtcccg cctcggcctc tcaaagtgct 62100

gagattacag gcgtgagcta ccctgcccag gctgaccatg tatattccta atttttgtac 62160

tttttttttt ttgagacgga gattcactct tgtggcccaa gctggagtgc aatggcatga 62220

tctcggctca ctgcaacctc tgcctcctgg gttcaagcaa ttctcctgcc tcggcctccc 62280

gagtagctgg gattacaggt gtgcaccacc acgcctgact gacaatatat attcttaaac 62340

agctcagatt ccattttgga tcctcccatc aggactggaa cttaggtgta ctggaactta 62400

ggtgaacact tggctcaaaa ttcattgctg ttctctataa atctagccag ttctcttggt 62460

taaatttaag gtatgtatag tagtcgctgc tttttctttc gggggacaaa actcaggagg 62520

attgcttctt gattcataag ggcaacctgt tgagattttc acgcaaggaa cccgagatgt 62580

tcatttaatg ggcttataat ttgggattcc agaacacatg caaacagggc aaatgaatgt 62640

ttggctgtat ctttattttt gtgttcattt cagcctggtc aaggttttag aatccaagga 62700

aaccaataaa cacccagagt gctggagcaa gactgtctcc tgctgtgacc ctcaagatgg 62760

aagcagtttc tgttgtctga aaggaaagaa agtgcttcct ttttgagggt tactgtttga 62820

gaaaagcaac cttgaggttg atgctgatgt ttgtaacaca cctgcagagt atacttataa 62880

tcagacttgg gtgatgtgag gttttgtttt tactccaaga tgagggtctc cacccaggct 62940

ggaatgcagt tgcccttcca acctagaact cctgggatga actggtcctc tatgcctcag 63000

cctgccgagt agctgggact ataatatagg ggtgtgccgc catatttggt agatttttca 63060

attttttgta gagatggggc ctcaccctgg ccaggatggt ctcaactcct gagctcaagc 63120

aatcctacag gtgtgagcca ccgtgcccat tttaaagata gttgacatga tcaagcatag 63180

tgggacacac agccccagct actgcagagg ctggggtggg agagtctctt gatttcaatg 63240

ctataccgtg cactaatgac acctttgaat agccactgca ctccagcctg ggccaacata 63300

gcaagatccc atctcttaaa aaaaaataca ttacatggca cctggttcca gagacaccat 63360

ttgtgttggt caaacaatgg cctcctataa atttagttta atgaatccaa accatgtttc 63420

cttcattaag agagtaaata agcctccaag tctatccagt ctttttgaga cagagcttgg 63480

ctcttgtcac tcaggctaga gtgcaatggt gcaatctcag ctcactgcaa cctctgcctc 63540

ctgagttcaa ggagttctct cacctcagcc tcccaagtag ctagaatgac aggcgcctgc 63600

caccatgacc agctaatttt tgtgttttta ctagagatgg ggtttcacca tgtcggtcag 63660

gctggtctcg aactcctgac ctccgtgatc cacccacctc agcctcccaa agtgccggga 63720

ttacagacga gagccaccac gcccggcctg tccagtcttg ttctgcagtc cccaatgggg 63780

attttttttt tcctgttcat ttgtcttttt attttaatct acttgtccct gacactgaaa 63840

atttttcctt cctaacagct taccatcatt tctcagaata gacctgcttc ctctgtgaga 63900

agcttatagt tgattcaacc ctcaaccact aatgccaaca accctagtga gttcttttga 63960

ctacagctgg accatttatc ctgtttcctg tgggtagcgg ttccaatgta ccattccaac 64020

aggcaaaacc tcgcctctga atacaggttg cttggcaaga tctaaaatgt ttgctgtgcc 64080

taatataaac tattgtaaag aaaatccatc tcaatcacag tgacaaatgt cacatgagac 64140

aaaaccacag atattttcgc aaaaataggg tcctttagaa ccctagaagg gtctctctag 64200

taacaggtgg gatgttcagc agctcttgtt gttgccacag tgagcgatgc ctgttcgtcc 64260

agcccttaac acctcttact ccatggaagt tctgcctgca ctgctttata gaacacctct 64320

tgggttgagg tagagttgga ggggacctca gtgtcccttg ctgatgggat gtgcactgct 64380

tagcaagcgc acggaggtgg agtgcatggg ctctgagttt ttattgggta aatgcagccg 64440

aaatgtagtg tgcatgaaca ggtcaaaaaa ttgcacattt gatttaattt ttaaatttta 64500

gagatggggg tctcactgtg tcacccagac taaactgggc tgtgctccta tgcgtaccct 64560

agtagctggg acttcaggtg aatattaacc catgcatagg caaggagaga ggaaggctct 64620

gacagtctgt gatctcccct cactgcaacc tccaccctct ggactgggaa cgtcagggca 64680

ctgcaccgat gcaggcagga tgagccgagg ggaaaggaga gccaggcatc actggctggg 64740

gacattttgg gtttgatctg gatggagcag gtgtctcctg gagagagagc ccctgggatt 64800

ttcactctgc tccctggctg tcttagtcat ggaatctgac aacagagact cctgcccagg 64860

gccacttcat ttggtttctg gaccccagtg gtccttcctg cctggactta ggatcttttg 64920

gggaagtttg ggatctggca gggcatctgc ataatccata gaaatccctg agagtcactt 64980

cccttggctg acatctccat gttcctaccc attaccttcc aaaggagacc cttaactgaa 65040

ttaccaaagg gggcttccca gagcagggaa acccggttaa ctttctattt caggtcaaca 65100

gtatacttga gatgtacttg aactagaaat gattggttgt ttaggtgtgg gcatttgttt 65160

ttcctaactt agtctccaag aaaaaaaatt attgaggttt tacagcctag cctgtggtct 65220

atcctgaaga atgttcgtgt gtgtgacaat gtacattctt tttttccccc cagctctgtc 65280

aaatcttact gaggagcttt tacggcctag cctgtggtct gtcctggagg gtatttgtgt 65340

gtacaacaat gtatattctt aaacattatc ttagattcca ttttggatgt tcccatcagg 65400

actgtgtgtt tctgtgctgg aactcaagtg aacactggct caacatcctt agaaatccag 65460

cccaattctc ttggttaaag ataaggtatg tgtggtaggc attgcttttt ctctttgggg 65520

acaaaactca ggaggattgc cccttgatga acaaggctaa cctgctaagc ctttgaagca 65580

aggaactgga gatggtcctt tcaggggttt atgttctgga ttccataaaa catgcaaaca 65640

ggggcaatga atgcaccttt tttattttta tttttatttt tttttgagat ggagtcttac 65700

tcttgccagt ctggagtgca gtggcacgat ctcggctcac tgcaacttct gcctcctggt 65760

tcaagtaatt cccctgcctc agcctcccga gtagctggga ctacaggtac atgccaccac 65820

aggcggcaaa tggttgtatt tttagtagag acggagtttc accatgttgg ccaggatggt 65880

cgtggtctct tgaccagcct cccaaagtgt tgggattcca ggcgtgagcc accgcgcctg 65940

gccaatgaat gcatctttat ttttgtgttc attttaatct ggtaaggtaa attccaacaa 66000

aaaacccaga gttttggagt gagaagatct catgcagtca ttctccaaaa gaaagcactt 66060

tctgttgtct gaaagcagag tgccttcttt tggagcgtta ctgtttgaga aaaaccacgt 66120

tgaagttgat gctgatcttg gtaacacatt tgcagagcgt gcttatcatc agacttgcat 66180

gatgttgggg ttctgttttt gtttagtttt tttgcaacac agggtctctt gcccaggctg 66240

gagtgcggtg acacttccaa cctagacctc ttgggctcag ttggtccctg cccccacccc 66300

ctcccccttt tttttctctt gagacagtct ctctctgtgg cccaggctgg agtgcagtgg 66360

tatgttctct gctcactgca acctctgcct cccgagtagc tggtattaca ggcacatgcc 66420

accacgcctg gctaattttt gtatttttag cagagacgga gtttcatcat gatggccaga 66480

ttagtcttga actcctgacc ttaggtgatc cacctgcctc ggcctcgcaa agtgctggga 66540

ttacaggcat gagccactgt gcctggccca caacgtatat tcttaaatat catcttagat 66600

tccattttgg ttgctcccat cgggactgtg tgtccctgtg ctggaactca agtgaacaca 66660

tggctcaaaa tccattgctg ttctctagaa atccagccca attctcttgg ttaaatataa 66720

ggtatgtgtg gtaggctttg ctttttctct ttggagacaa tactcaggag ggttgcccct 66780

tcgtgaacaa ggctaacctg ctgagccttt gaagcaagga actggagatg gtccttttag 66840

gggtttatgt tctggattcc agaaaacatg caaacaggga caatgaatgc atctttattt 66900

ttgtgtccat tttaacctgg taaggaaaat tccaacaaaa acccagagtt ttggagcgag 66960

aagatctcat gcagtcattc tccaaaaggg agcactttct gtttgaaaga aaacaaagtg 67020

cctcctttta gagtgttact gtttgagaaa aaccacgttg aagttgatgc tgatcttggt 67080

aacgcatttg cagagcgtgc ttatcatcag acttgcatga tgttggggtt ctgtttttgt 67140

ttagtttttt tgcaacacag ggtttctgtt gcccgggcag gagtgcggtg gcgcttccaa 67200

cctagacctc ttaggctagt tggtcccccc tacttttttt gttgtttttg ttcttgagac 67260

agagtctcac tatgtgggcc aggcgggcag gcagtgacac attctctgct cactgcaacc 67320

tcggcctccc aagtagctgg tattataggc acgtgccacc acgcctggct aatttttgta 67380

tttttagcag agatggagtt tcaccacatt ggccaggtta gtcttgaact cctgacctca 67440

ggtgatctgc ccgcctctgc tttccaaagt ggtgggatta caggcatgag ccaccctgct 67500

cggactgcag ggtgtttttt tttttttttt taattattat ttgtattttt ttgtgctgcc 67560

aaagcaagca cttgtgtgta ggaattttgt ttgtttgttt gtttgtttgt ttgagatgga 67620

gtctcattct tgtcgcccag gctggagtgc agtggcatga tctcggctca ctgcaaccta 67680

cgccttccgg gttcaagcga ttctcctgcc tcagcctcct gagtagctgg gatttcaggt 67740

gcatgccgcc accccggcta atttttgtcc ttttggtaga atcggggttt tgccatgttg 67800

gtccggctgg tctcaaactc ctgacctcaa ctaatccacc tgcctcggcc tcccaaactg 67860

ctgggattac aggcatgagc caccgcaccc ggtgaggagt tatttttaat gtgagcaaac 67920

agtatattct tttttttttt tttcatagac agggtctcaa caatatatga tgtatattta 67980

atcatatagt cttatcatgt atataatgta gtcatatgta caccaaaccc tgttctacac 68040

acggagaata ctcttgtatc actttgggta tttatttact tgtttctttc tgtgtttttc 68100

ttgtttgttt gtttgttttg ttttgagaca aggtctagct ttatcaccca ggctggagtg 68160

cagtggtgcc atgtcgactt actgcaacct ccacctgcca cctcagcctc ctgagtactt 68220

gtctacaggt gcgcaccacc acacccagct aacttttcta ttttttgtgc ataaggtttc 68280

accatattac ccatgctggt tgagctcgaa ctcctgagct caagtgatcc tcctgccttg 68340

ccctctttaa gtgctgggtt tattggtgtg agccaccacg ccccacccat acttgtgtat 68400

ttctgtattt attgaggagc ttatacagca taccctgtgg tctatcctgc aggatgtttg 68460

tgtgtgagac agtgtatatt cttgaacata gtagattcca ttttggatgc tcccatcggg 68520

actgtgtgtc cctgtgctgg aactcgagtg aacacttggc tcaaaatcca ttgctgttct 68580

ctagaaatcc agctcaattc tcatggttaa atataaggta tatgtggtag gcattgcttt 68640

ttctctttgg ggacagaact caggaggatt gccccttgat gaacaaggct aacctgctga 68700

ttctttgaag caaaggactg gagatggtcc ttttaggggt ttatgttctg gattccagaa 68760

aacatgcaaa cagggccaat aaatgcatct ttttgttttg ttttgttttg tttttgagat 68820

ggagtctcgt tctgtcaccc aggctggagt gcagtggcac aatcttggct cactgcaagc 68880

tcagcctcct gggttcacgc cattctcctg cctcagcttt ccaagtagct ggggctacag 68940

gtgcccacca ccacgctagg ctaatttttt gtatttttag tagaggcggg gtttcaccct 69000

gttagccagg atggtcttga tctcctgacc tcgtgatctg cccgcctcgg cctcccaaag 69060

tgctgggatt acaggcatga gccactgcgc ccggccccaa taaatgcatc tttatttttg 69120

tgtccattta aacctggtca aggaagattc ccacaaaaaa tccacggtgc tggagcaaga 69180

ggatctcagg ctgtgaccct ctaaagggaa gcgctttctg tggtcagaaa gaaaagcaag 69240

tgcttccttt tagagggtta ccgtttggga aaagcaatgt tgaagttgat gctgatcttg 69300

gtaaaatatt tgcagagcgt gcttatcatc agacttggat gatggtgggg ttttgctttt 69360

gttttgttgt attccaagac aaggtccctg ttgcccaggc tggagtgcgg tgacacttca 69420

acctacattt cttgggctcc ggtcgttttt gtttgtttgt ttgtttgttt gacagggagt 69480

ctcactgtgt gtctcagcaa tgcagtggca ctatcttggc tcactgaaac ctcagcctcc 69540

tgagtagctg ggatcacagg tgcgtgcaac cacgcccatc taatttttgt attttttgca 69600

ttttcagtag agacggggtt tcaccgtgtt agccaggatg gtctcgatct tctaacctcg 69660

tgatccgccc gcctcggcct cccaaagtgc tgggattata ggcgtgagcc accgcgcccg 69720

gccgagaaac taatcttttg agatgaactc tgagatgtgg attttagctt gtttgcagcc 69780

accaccactc tagttttgga agattttcat caccccgaag aggcttatac tcatttgcag 69840

tcagtaccca cccacctctt ccacccagac cgtggcaacg actccccatc tctctagctc 69900

tggatctgcc tcttgtaggc cggtcacgta gaccaatctt gtatgggtgt ccagttgagg 69960

ataatgggtt ggtcctggtt gtctgcaatg tgaatcttac cactgaaggg tggtccctgg 70020

agggaagcag gaggctggga gaactgggcg gaacatcctt tgggaatgga gtggggcggg 70080

cagaccctga tgtctgggaa gctcacaagg gtggaagacc ccatcttcct ccctgagaac 70140

tgcaaggtga ccctcctggg gcactggaag gagtgaaggc ctctgggctg ggaacgtcag 70200

ggcactgcac cgatgcaggc aggatgagcc gaggggaaag gagaagcagg catcattctc 70260

tggggacatt ttgggtttga tctggatgga gcaggtgtct tctgggagag agagcccctg 70320

ggattttcac tctgctccct ggctgtctta gtcatggaat ctgacaacag agactcctgc 70380

ccagggccac ttcatttggt ttctggagcc cagtggtcct tcctgcctgg acttgggatc 70440

ttttggggaa gtttgggatc tggctgggcg tctgcataat ccatagaaat ccctgagagt 70500

cacttccctt ggctgacatc tccatgttcc ttacccatta ccttccaaag gagaccctta 70560

tctgaattac caaagggggc ttcccagagc agggaaacct ggttaaattt gtatttcaga 70620

ttaacagtat acttgagatg tacttgaagt agaaatgatt ggctgtgggt gtcggcattt 70680

gttttttcta attttaaata tgggaccatc atgaatttgg gtgtcacttt gtgcagggga 70740

cgtgggaatc gctgtcattt tttttttttt ttaacatatg ctgccagaga aggcacttag 70800

gtggagaaat tactcttcgt gtgagcattc agtatataga acttcctttt ctgggggcag 70860

tgtcttatat atgatacaca ttttatcata tcatcttcta tagataatat aatgattaac 70920

acaaaacact cttctacaca cacagaatac tcttgtatca ctttgggttt tttttccttc 70980

gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt gttatgagac aaggtctggc 71040

tctgtcaccc aggctgcagt gcagtgttgc agtcttggcc catgccccac cgcagcctct 71100

ggaatagctg tctgcaggcc aggcacggta gctcaggcat gtcgtctcag cactcttgga 71160

gaccaaagca ggctaatcac ctggagtcag gagtttgaga ccagcctggc caagatggtg 71220

aaacctcatt tctactaaaa gtacaaaaat tagccgggcg cggtggctca cgcctgtaat 71280

cccagctctt agggaggcag aggcaggagg atagcttgag cccaggagtt tgagacctgc 71340

ctgggcaata cagtgagacc ctgttctcca caaaaagaaa agaaaaaaaa gaaaaaaaaa 71400

ttttaaaaat gcaaaattag gtgggcatcg tggcacgtgc ctgtaatccc aggtacccca 71460

gaggctgaga caggagaatt gctggaaacc tggaggcaga gcttgcagtg agctgagatc 71520

acggccctgc actacagcct gggcaacaaa gtgagacttg tttcaaaaaa aaaaaaaaaa 71580

tttattgagg agcttttatg gcctagcctg tgttctatcc tggagaatgt gtgtacaaca 71640

gtgtatattc ttaaacatca tcttagattc cattttggat gcttgcatgg ggactgtgtg 71700

tccctgtact gggacttatg tgaacacttg gcttcaaatc cattgctgtt ctctagaaat 71760

ccagcccaat tctcatggtt aaatataagg tatgtgtggt aggcattgct ttttctcttt 71820

ggggacagaa ctcaggagga ttgtctcttg atgaacaagg ctaacctgct gagcctttga 71880

agcaaggaac tggagatggt ccttttaggg gtttatattc tggattccag aaaacatgca 71940

gagagagtca ataaatgtat ctttattttt gtgtccattt taaccaggtg aggaaaattc 72000

cgtcaaaaaa cccagacttt tggagcgaga agatctcatg cagtcattct ccaaaagaaa 72060

gcactttctg ttgtctgaaa gcagagtgcc ttcttttgga gcgttactgt ttgagaaaaa 72120

ccacgttgaa gttgatgctg atcttggtaa cacatttgca gagcgtgctt atcatcagac 72180

ttgcatgatg ttggggttct gtttttgttt agttgttttg catcatgggg tctctgttgc 72240

ccaggctgga gtgcggtggc gcttccaacc tagatctctt gggctcaagt ggtcccgtgc 72300

ttgctttttt tttttttctt gaaacagagt ctctgtctct ggcccaggct tgagcgcagt 72360

ggcatgttat ctgctcactg caacctctcc tcccaagtag atgggattac aggtgcacgt 72420

gccacgacgc ctggctaata tttgcatttt tagcagagat gcagtttcat catgatggcc 72480

aggttagtct ccaactcctg acctcaggtg atccgcctgc ctcagcctcc caaaatgctg 72540

agattatagg tgtaagccac ccagcctagc ctccaatttt ttttaatgtg tgctgccaat 72600

gcaagcacat atgtgtagga attatttttc atgtgagcac atagtatatg gaacttaatt 72660

tttattgatg tacggtctca aaaatatata ttttatcata ttcttattat acatgtaaca 72720

tagtcatata cacacaaaac actcttctat acacagagaa tactctcgta tcactttggg 72780

taattttttg tttgtgtgtg tatgtttttg tttttttttt agataaggtc tgggtctgtc 72840

acccaggcta gagtgcagtg gtgccatctc agcttattgc aacctccacc tgccacttca 72900

gcctcttgag tagctgtcta caggcgcaca ccaccacgcc cggctaactt ttctgttttc 72960

tctacacatg aggtatcacc atgttgccca ggctggtctc aaactcttga actcaagtga 73020

tactcctgcc tcagcctcct aaagggctgg gtttagaggt gtgagccacc acacctcacc 73080

tgtacctgtg tgtttctgct cattttattg aggagctttt acagcctggc ctgtggtctc 73140

tcctggaggg tgtttgtgtg tgtgacaatg catattcttc aacatcgtag attccatttt 73200

ggatgccccc gtcgggactg tgtgtcactg tactggaact cgagtgaaca cttagctcaa 73260

aatctattgc tgttctctag aatccagccc aattctcatg gttaaatata aggtatgtat 73320

agtcggcatt gctttttgaa acaaggaact ggagatggcc ctgatagggg tttatgttct 73380

ggattccaga aatcatgcaa acagggccaa taaatgcatc tttatttttg tgtccatttt 73440

aacctggtca aggaaaattc caacaaaaaa tcaatggtgc tggagcaaga agatctcagc 73500

ctgtgaccct ctagagggaa gcgctttctg ttgtctgaaa gaaaagaaag tgcatctttt 73560

tagaggatta cagtttgaga aaagcaacgt taacgttcat gctgatctcg gcaatacatt 73620

tgcagagcgt gcttatcatc agacttggat gagggtgggc ttttgttcct ggtttgtttg 73680

ttttctaaga cggggtctct tttgcccagg ctggagagca gtggcacttc caacctagct 73740

ctcttgggct caggtggtcc tctttgcagg gtcacgatct ctgctcacta caatctcccc 73800

ttcccgggtt gaagtggttc tcccatctca gtctcctgtg ttgctgggag tacaggccca 73860

tgccaccacg cccggctaat ttttgtattt ttagcagaga tggggtttca ccgtgttgac 73920

caggatggtc tccatctcct gacctcgtaa tccacctgcc tcggcctccc aaagtgctgg 73980

gtttacagtc atgaaccact gcacccggcc tttgatctta attttcaata tgagacccat 74040

catgaatttg ggcatcacct ctgcacaggg gccatggtga tctctgtcac tccattcatt 74100

tcttatgtgt ggagcaaaca tttatgtgta ggagtaattc tttatgtgag catacagtat 74160

atggaacttt cttttttttt gagatagggt ctggaaaata aatgatgtac agccaggcgc 74220

atgagtatat gagcagtttt tgtgtgtata cgacatgagc ggtggctcat gcctgcaata 74280

tcaacacttt ggcaggttga ggcgagtgga tcacttgagg tcaggagttt gagaccagcc 74340

tgaccaacat ggtgaaaccc tgtctctact aaaaatacaa aaaattagcc agctatggtg 74400

gcgcacgcct gtattcccat ctgctgggga agttgaggga ggagaatcac ttgaacggac 74460

agatggaggt tgcagtgagc caagatcacg ccaccacact ccagcctggg cgagagaata 74520

agactgtctc aaaaaaaaaa tatatatata catatatatt tatatagaga gagagactct 74580

gtctcaaaaa aaaaaatata tatatgggtg tgtttgtgtg tatatatata ttatatatga 74640

tgtatatttc atcatatagc catatatata attatataca caaaaaatac tcttctacac 74700

acagagatga ctgatatcac tttggggtgg tttttgcttg tgtgttcttt taaatttttt 74760

tttgcaacaa gatctggctc agtcgcccag gttggagtgc agtgtttcag tcttggctca 74820

ctgcaacctt cgcctcccac cccagcctcc tgaattagtg tctacaggca tgtaccacca 74880

cacctggcct acatttgtat ttttttacag acagggtttc accgtgttgt acacgctggt 74940

cttgagctcc tgagctcaag tgaatctcct gccttggcct cctgatgtgc tgggattaga 75000

ggtatgagcc accacaccca actggtactt gtgtattcct gttcaaattt tattgaggag 75060

cttttaccac ctagcctgtg gtctatcctg gaggatgttt gtgtgtgcga caatgtatat 75120

agttcaacat cttagatcac attttggatg tttccaatgg gactgtgtgt ctccgcactg 75180

gaactcaaat gaacacttgg ctcagaatcc atttgctgtt ctctggaaat ccagtccaat 75240

tctcttggtt aagtataagg tatgtctagt aggcattgct ttttctgttt gagaacaaaa 75300

ctcgggagga ttgtcccttg atgaacaagg ctaacctgct gagcctttga agcaaggaac 75360

tggagatggt ccttttcagg cgttttattc tggattccag aaaacatgca aacagggcca 75420

ctaaatgcat ctttattttt ctgtccattt aaacctggtc aaagaaaatt ccaacacgaa 75480

acccagagtg ctggagcaag aagatctcaa gctatgagtc tacaaaggaa agcgctttct 75540

gttgtcagaa agaagagaaa gcgcttccct tttgagggtt acggtttgag aaaagcagtg 75600

ttgaagttga tgctgatctt ggtaatacat ttgcagagca tgcttatcat cagacttgga 75660

tgatagcggg gttatgtttt ggttttgtgt ttttctaaga cagggtctcc gttgcccagg 75720

ctggagtgcg gtggcacttc caacctagct ctcttgggct caagtgatcc tctttttatt 75780

tatttatgta ttcatttttg agatggagcc tggctctgtc acccaggctg gagtagagtg 75840

ggatgatctc cactcattgc atcctctgcc tcccaggttc cagaaattct cccacctccg 75900

tgtcactagt agctgggatt acaaatgctc accactatga cctgccagtt tttgtacttt 75960

tggcacagac ggggtttcac catgtttgcg gggctggtct caactcctga cctcaagtga 76020

tctgcctcct tggcctccca cagtgctggg gattatagat ataagccacc gtgcccggcc 76080

cattcctgtc tttttaattt aatctgctcg tccctgacat cgaaaatttt tcttgcttaa 76140

cagcttccac ttatttctcg gaatagacct ttttctgcag gaagggtgta gttgattcaa 76200

ccctcaccca ctcatgccaa ccgtagtgag tgctgtgact gcagctgcac tgtttatcct 76260

gttttctgca ggtaattttt ccagtgtacc attccaacct gcaaaacctc acatttaaat 76320

gcaaatttct tggtaagctg tgaaatgttg gctgtggcta atatgagaaa caagctatta 76380

tagacaagat gaatctcagt tgcagtgata aatgtcacat gggacaaaac cacagatact 76440

ttcacaaaaa ccttgaggac cctagaaggg cctccctagt aacaggtggg atgcgcatcc 76500

gctcttgttg ccatagtgag tgatgcctgt tcgtccagcc ctcaacacct ttcactccgt 76560

ggaagttatg cctgcactgg tttacagaac ctcccttgac ttgacttgag gtagagttga 76620

agggaacctc agtgtccctt gcagatggga tgtgcattgc ttcgcaagag cacagaggtg 76680

gagtgcatgg gctttgagtt tttattgggt aaatgaagct gaaatacagg gtgtatgacc 76740

acgttatcaa cgcacacttg atttaggttt tgtattttag agacgggggt gtgagctgcc 76800

aaagcaagga cttatgtgta ggaattattc tttctgtgag catacagtat atggaacttt 76860

ctttcttttg agacagggtc tcaaaaatat atgatgtatg gccaggtgcg gtggatcacg 76920

cctgtaatcc cagcagtttg ggaggccgag gcgggtggat aacctgagat caggagggcg 76980

agaccagact ggccaacatg gtgaaaccat gtctctgcta aaaatacaaa aaattagcca 77040

gggctggtgg cgcacgcctg tagtctcagc tactccggag ggtgaggcag gagactatca 77100

ctttgagtgt gtgtctttgt gtgtgtgtgt gtgtgtgtgt gtgtgtgtgt atgtgtttct 77160

agtttttaca agacacagcc tgactatatc accgaggctg gagtgcagtg gtgcaatgtt 77220

ggctcactgc aacctccacc tgccaccgca gcctcctgaa ttagtgtcta caggtatgca 77280

ccaccacacc tggctcactt ttgtattttt tgtacagatg aagcttcacc atgttgccca 77340

ggcttgtctg gaactcttga gctcaagtga tcctctcact tcggcgtcct gaagtgctgg 77400

ggttagagat gtgagccacc gcacccaaca ggtacttgtg tattttttta ttgaggagct 77460

tttacagcgt agcctgtggt ctctcctgga ggatgtttgt gtgtgcgaca atgtacattc 77520

ttcaacatct tagattccat tttggatgcc cccatgggta ctacatgtgc ctatactgga 77580

actcaagtga acacttggct caaaatccat tgctgttctc tagaaattca gttcgattct 77640

cttggttaaa gataaggtat gagtagtagg cattgctttt tctctttggg ggcaaaactc 77700

aggaggattg ccccttgatg aacaaggcta acctgctgag cctttgaaag aaggaactgg 77760

agatggtcct tttagggggt ttatattctg gattccagaa aacatgcaaa cagggccagg 77820

aaaaatgcat ctttattttt gtgtccattt aaacctggtc aaggaaaatt ccaacaataa 77880

acccagagtg ctggagcaag aagatctcag gctgtgaccc tccagaggga agtactttct 77940

gttgtctgag agaaaagaaa gtgcttccct ttggactgtt tcggtttgag taaagcagcg 78000

ttgaagttga tgctgatctt ggtaatacat ttgcagagca cgctcatcat cagactcgga 78060

tgatgttggg gttctgattt tgtttttctc caagacaggg tctctgttgc ccaggctgga 78120

gtgcagtggc agttccaacc tagctctcct gggcccaaga gaccctctat ttatttattt 78180

atttatttat ttatttattt atttatttat tcattcattc atttagagat ggaatctggc 78240

tctgtgaccc aggctggagt gcagtaggac aatctccact cactgcaacc tccatctccc 78300

aggttccagc agttctgcca cctcagcctc ccatgtagct gggattacag gcgcccacca 78360

ccatgtcatg ctaatttttg tatttttagc agagatgggg ttacaccatg tttgctgggc 78420

tgttctcaac tcctgacttc aagtgatttg cctccctggc ttcccaaagt gctggtatta 78480

caggcgtgag ccactgtgtc cggcctcaag tggtcttcct gagtcagcct cccaagtagt 78540

tgggattaca tggggcgtga cacaacactt ggttcagctt ttaatttttg gtagagatgg 78600

ggtctctgtt gctcaggacg gtctcaactc ctgagctcaa gcgatcctac aggtgtgagc 78660

caccttgtcc tgatgaccca tttcaaagat agttgacttg gccaggcatc atggggcaca 78720

cagtcccagt tactgcaggg gccggggcgg gagggtgctt tgattttaag gctataccgt 78780

gcacagatcc cacctttgaa tagccactgc actccagcct gggccaacat agcaagatcc 78840

catttcttta aaaacagatt acatagcacc tggtcccaca gatttcattt gggttggtca 78900

tgtcatacaa tggcctccca tcaatttagt ataatcaatc caaaccatgg ttccttcatt 78960

aagagagtgg gtaatcctcc aagtccatcc agtctattct gtaatcccca gtgggtgtcc 79020

gtattgatac aatttctttt tgttcctgtt cactcgtgtc tttttagtgt aatctgcttg 79080

tccgtaacac tgaaatttgt ttttccccaa catcttgcca tcatttctcg gaatagacct 79140

gctttctctg caggaagggt gtagttgatt caaccctcac ccactaatgc caaccccagt 79200

gagttctttg actgtagctg ccatgtttat cctattttct ttgggtaacg attccaaggt 79260

accattccaa tgggcttaaa ccttgcatct aaatgccagt tacttgctaa gatgtcaaat 79320

gtgtcttata ttagctgtgg ctaatataag aaactgttgt agacaagatg aatctcagtc 79380

acagtgataa atgtagcatg caacaaaacc agttgttttc acaaaaatct tgaggaccct 79440

agaaggggct ccctcgtaac aggtggaatg cacagcagct cttcttgttg ccatagtgag 79500

cgatgcatgt tcttccagtc ctcaacacct tttatttatt tatttattta tttatttatt 79560

ttattattat tatttttttt tgagacagag tctcgctctg tcacccaggc tggagtgcaa 79620

tggtgtgatc tcagctcact gcaaccactg cctcccgggt tcaagcgatt ctcttgcctc 79680

agcctcacgt gtcgctagga ctacaaccgt gcaccaccac gcccggctaa tttttgtatt 79740

tttaatagag acagggtttt gccatgttgg ccaggttggt ctggaactcc tgacctcagt 79800

tggtccacct gcctcagcct cacaaagtgc tgtgaataca ggtgtgagcc aacacgcccg 79860

gctatcaaca acttttccta cgtagaaatt attcgggcac tggtttatag aacctcacgt 79920

gggttcacgt agaattgaag gggacctcag cgtcccttgc agatgggatg tgcaatgtat 79980

ttgagatgta ctcgaacatt gctgttgagg tgtgggcatc ttttgctttt tcctaatttt 80040

aaatatggga ctagtctggg tatggtgact tccaccagta attctagcac tttgggaggc 80100

tgagacagga ggatcacctg aggtcagttg ttcgagacca gtctggccag catggtgaaa 80160

ccccgtctct actaaaaata caaaaattag ccgggtgtgg tggcactcac ctgtaatccc 80220

agctactcgg gaggctgagg caggagaatc acatgaatct gggaggcgga ggttacagcg 80280

agcagagatc acaccgttac actcctgcct gggcaacagt gtgagactct gtctctaaat 80340

aaataaataa ataaaaataa aaataattag gtgaatatgg gaccagcatg gacttgggtg 80400

tcgcctttgt gcagtggcca ggatattcta tgtcatttaa atttttttat gtgtacttcc 80460

aaagcaagca gttatgtata ggaattattc ttcctataag catgcaatat ttggaacttt 80520

ctttgagaca gggtctcaaa aatacatgat gtatatttca ttttatattc ttatatatta 80580

tgtacacaca aaatactctt ctaaacacag agaatactcc gatatcacct agggtgtgtg 80640

tgcgtgtgcg tgtgtgtgcg tgtgtgtgtg gttcttttct gagacaatat ctgcctctgt 80700

cacccaagat ggagtgtggt ggtgcaatct cggcttactg caacctccac ctcccaccta 80760

agcctgctga attagtctct acaggcatgc accaccacac ctggctaact tctgtaattt 80820

ttgtacacat ggggtctcgc catgttgccg gggctggtct cgagctcctg aggcaggtga 80880

tcctcctgcc tgggcctcct aaagttcgag gagttagagg tctgagccac tgggccccac 80940

ttgtacttgt gtatttctgc tcaaatttta ttgagtagtt tttacatcct agcctgtgat 81000

ctactctgga gagtgtttgt gcatatgaca gtgtatattc ttcaacatcg tagactccat 81060

tttggatgct cccttcgtga ctgtgggtcc tgtactggaa ctcgagtgaa cacttggctt 81120

aaaatccatt gctgttctcc agaaatcctg cccaattttc tcggttaaag gtacgtgtag 81180

taggcattgc tttttctctt tggggacaaa actcaggagg attgcccctt gatgaacaag 81240

gctaacctgc tgattctttg aagcaaggaa ctggagatgg tccttataga gttttatatt 81300

ctggattcca taaaacatgc atacagggtc aataaatgca tctttatttt tgtgtctatt 81360

ttaacttggt caaagaaaat tccaggaaaa aatccacggc atcagagcaa gaagatgtca 81420

ggctgtgacc ctcttgaggg aagcactttc tgttgtctga aagaagagaa agtgcttcct 81480

tttagaggct tactgtctga gaaaagcaac gttgtagttg atgctgatct ttgtaatatc 81540

tttgcagagc acgcttataa tcagacttgg atgatgttgg ggttttgttt ttcttttgtt 81600

tttttatcta agactgggtc tctgttgccc aggctgaagt gcagtggcac aatcttggct 81660

caccacaacc tccgcctccc gggttcaagt gattcttctg cctcagcctc ccaagtagct 81720

gggattatag gactgcgcca ccatgcctgg ctaattttgt atttttagta gagacagggt 81780

ttctttagga gcttttaagg cctcccttgt agtctatcct ggaggatgtt tgtgtgtgca 81840

acaatgtata ttcttcaaca tggtagattc cattttgact gctctaatca ggactgtgtg 81900

tccctgtgct ggaaatcaag tgaacacttg gctgaaaatc cactgctgtt ctctagaaat 81960

ccagcccaat tctcttggtt aaatataagg tatgcatagt aggcattgct ttttctttct 82020

ggacacaaaa ctcaggagga tttccccttg atgaacaagg ctaacctgct gagcctttgt 82080

aggaaggaac gggagatggt ctttttaggg gtctatgttc tggattccag aaaacatgca 82140

aacagagcca ataaataggt ctttattttt gtctctattt taacctggtc aaggaaaatt 82200

tctacaaaaa acccagggtg ctggagcaag aagatcccat gctgtgaccc tctagaggga 82260

agcgctttct gttgtctgaa agaaaagaaa gtgcatcctt ttagaggttt actgtttgag 82320

gaaagcaaca gtgaagttga tgctgatctt ggtaatacat ttgcagagca tgcttatcat 82380

cagacttgga tgatggtggg attctgtttt tattttgttt ttttttctaa gacagagtct 82440

ctcttgccca ggctggagta gggtagcact tccaacgtag atcttttagg ctcaaacggt 82500

cctctttttt tttttttttt ttttgaggcg gagtcttgct ctgtgaccca gacgagagtg 82560

caatggcgcg atcttgtgtg caagctcacc tcctgagttc aagcaattct tctgccccag 82620

cctcccaagt agctgggact acaggtgcct gccaccactt ccagctaatt tttgtatttt 82680

tagtagagac agggtttcac cacgttggcc aggctggtct caaactactg acctcaagat 82740

ccacccacct cagcctccca aagtgctggg attacaggcg tgggccattg tgcccagcgg 82800

tcctcttttt tgagatggag tcttgctctg tgacccaggc tggagtgcag tggcaggaac 82860

ttcgctccct gtaccctcca cctcccagat tcaagccatt ctctcctgag tcagcctccc 82920

tagtagctga gatcgcagac atgagccacc atacccagct aatttttgta tttttattat 82980

ttatttattt actcatcgat ttttgaaaca gagtcttact ccgtccatca ggctggaggt 83040

cagtggcaca atcccacgtt caagcaattc tcctgcctca tcctcccgag tagctgggat 83100

cacaggcgca caccaccacg cccggctaat ttttgtgttt tttttgtttg ttcgtttttt 83160

ttttttttgt agagacaggg tttcaacatg ttggccagcc cagtctccaa ctcctgacct 83220

taagtgagcc catacacaat caagaagaga gcgagacctg ttcttttttt ctgtctttga 83280

ctatttgaga cagtcttgct ctgtcacccg agatggagta cattagtgtg atcttggctc 83340

actgcaacct ctgcctcctg gactcaagca attctctttt tttttttttt ggagaaagag 83400

cctctctgtg tctcccaggc tggactgcag tggtgtgatc tcagctcact gcaacctcag 83460

ccttccaagt agctgggatt gcagacatgt acccccatac cagctatata tatatatgta 83520

ttttagacac agtttcacca tgttagccag gccattcttg aatcctgacc tcaggtgatt 83580

tgccgacgta ggcctcccga agtgctggga ttacaggtat gagccactgc actcagcctt 83640

ttttcctaat tttgaatatg ggacccatta tgaatttggg tgtcacctat gtgcgggggc 83700

tatggaaatc tctgtcaccc cattcatttg tgtgtgctgt caaagcacca atatgtggga 83760

attatgcttt atgtgagcat atagtatatg gaactttggt ttttttttat acagggtctc 83820

aaaaaaaaat gagtatattt catcacaatg ttctgtgtat aattccatac acacaaaata 83880

gtcttctata cacagagaac actcttatat cactttggag ttgtttttgt ttgtgtattt 83940

tgttttaatt tttatgagac agggcctggc tatgtcgctg aggctggagg gcagtggtga 84000

gatcttggct cactgcaacc tccacctgcc acctcagcct cctggattag tgtctacagg 84060

caggcaccac cacccctggg taagttctgt atttttggta ctgatgaggt ttcagcacgt 84120

tccacaggct ggactcttaa ctcctgcgat caagtgatcc tcccgctgtt caaattttac 84180

tgaggagctt ttaaggcctc gcctgtggtc tgtcctgtag tgtccctgtc ctggaactca 84240

agtgaatact tggtttaaaa ttgattgctg ttctctagaa atccagccca attctcttgg 84300

ttaaatataa ggtatgtgta gtaggcactg ctttttcttt ctggaggcga aactcaggag 84360

gattgcccct tgatgaacaa ggctaacctg ctgagccttt gaagcaagga actggagatg 84420

gtcgttttag gggtttatgt tctggattcc agaaaacatg gaaacagggc caataaatgc 84480

atctttattt ttgtgtccat tttaacccag tgaaggaaga tttcaaccaa aaacccacgg 84540

tgctggagca agaagatctc aagctgtgac tctccagagg gatgcacttt ctcttatgtg 84600

aaaaaaaaga aggcgcttcc ctttagagcg ttacggtttg ggtaaagcaa cgttgaagtt 84660

gatgctgatc ttggtaatat atttgcagag catgcttata attaagactt ggatgatggt 84720

gggtttctgt tttgtttttt gttttaaatc agagtctcac tctgtcaccc aggctggagt 84780

gcaatggcgc aatctcggct cactgcaact tccccctcct gggttcaagt gattctcctg 84840

cctcagcctc ctgagtagtt aggattacag acatgcgcca cctcacccgg ctcagttttt 84900

gtatttggtg gagatggggt ttcaccatgt tagtcaggct ggtcttgaac tcctgatctc 84960

aggtgatcca cctgcctcag cctcccaaag cattgggatt acaggcgtga gccaccacgc 85020

caggcctgtt caacatctta gatttcattt tggatgttcc tgtgaggact gcgtgtccct 85080

gtgctggaac tcaagtgaac gcttggctca aaatccattg ctgttctcta gaaatccagc 85140

caaattctat tggtgaaata taaggtatgt ctagtagtca ttgctttttc tctttggaga 85200

caaaactcag gaggattgcc ccttgatgaa caaggctaac ctgctgagcc tttgaaacaa 85260

ggaactggag atggtccttt caggggttta tattctggat tccagaaaac atgcaaacag 85320

ggccagtaaa tgcatcttta tttttgtgtt cattttaacc tggtcaagga aaattgctac 85380

aaaaaaccca gggtgctgga gcaagaagat ctcatgctgt gaccctctag agggaagcgc 85440

tttctgttgt ctgaaagaaa agaacgcgct tccctataga gggttaccct ttgagaaaag 85500

cagcattgaa gttgatgctg atcttgctaa tacatttgca gagcatgctt atcatcagac 85560

ttggatgaag gtggggttct gttttttttt tttttctaag acaggatctc tgttgcccag 85620

gctggaatgt ggtggcactt ccaacctagc tctcttgggc tcaaatggtc ctctcttttg 85680

ggatagagtc ttactctgtg acccaggctg gagtgcagtg gcgtgatctc agcttactgc 85740

cacctccacc tcccagtttc aagccgttct cctgcctcag attcctgagt atctgggatt 85800

ataggtacct gacaccacgc ctggctaata cttgcatttt tctttctttt ttcttttctt 85860

tttttttttt ttttttttga gatggagtct tactctgtca ccaggctgga gtgcagtggt 85920

gcgatcttcg cttatttcaa cctccgcctc ccagtttcaa gtgattctcc tgcctcagcc 85980

acccgactaa ctaggatttc aggcatgcac cacacgcctg gctaattttt tgtattttta 86040

gtagagacag ggtttcacca tgttggtcag gctggtctcg aactcccgac ctctggtgat 86100

cttcccgcct tggcctccga aagtgctggg attgcaggca ttagccaccc cgcccggcca 86160

ttatatattc ttatatatat aataccatat acgtgcaaaa tactcttcta catgaagaga 86220

atactcttat atcactttgg ggtttttttt gtttgtttgt gtgtgtgtga gtgtatgttt 86280

tatttgtttt tggataaggt ctggctctgt cacccagtct agagtgcagt tgtgccatct 86340

ctctcagctc actgcgactt ctacctccca cctaagcctg ctgaattagt gtctacaggc 86400

atgcaccacc acacctggct ccgttttgta ttttttatac acatgggctt acaccatgtt 86460

gccgagactg gtctcgaaca cctgagctca ggtgatactt cagtttcagc ctcctaatgt 86520

gctgggatta gaggtgtgag ccaccgtgcc ccacttgtag ttgtgtgttt ctgttcaaat 86580

tttattgagg agtttttagg gcctagcctg tgatctaccc tggaggatgt ttgtgtgtac 86640

aacaatgtat attctttaac atcgtagatt ccattttgga tgctcccatc gggactgtgt 86700

gtccctgtgc tggaactctg gtgaacacct ggctcaatat ccatttctct tctctagaaa 86760

tccagcccag ttctcttggt taaatataag gtacatatgt ctatgaggca ttgcttttgc 86820

tctttgagaa caaaactcag gaggattgct ctttgatgaa caaagctaac ctgctgatcc 86880

tttgaagcaa ggaactggag atggtccttt taggggttta tattctggat tccagaaaac 86940

atgcaaacag ggccaataaa tgcatctttg tttttgtgtc cattttaacc gggtgaagga 87000

aaattccaac aaaaaaccca gagtgttgga gcaagaagat ctcatgctgt gaccctctag 87060

agggaagcac tttctcttgt ctaaaagaaa agaaagcgct tctctttaga ggattactct 87120

ttgagaaaag caacgttgaa gttgatgctt atcttggtaa taaatttgca gagaatgctt 87180

ataatcagac gtggatgatg ttggtgtttc atgtttgttt tgttttgttt ttaatacagg 87240

gtgtctgttg cccaggctgg agtgtggtgg tccctctcca acctagatct cttgggttca 87300

agtggccctc ttttttgggt cagagtcttg ctctgtggcc ctggctggag tgcagtgtca 87360

ggatctctgc tcactacaac ctctgcctcc tgggtaaaag cgattctcct gcctcagcct 87420

cccaagcaac tgggattaca ggcatgtgct accatgcccg gctaattttt gtaatttcct 87480

ttatttattt ttgagacaga gtcttactct ttcgcccagg ctggagtgca gtggtgcaat 87540

ctcggctcac tgcaacctcc aactcccacc tcagcctcct gaatagctgt ctacaggcat 87600

gcaccaccac acctggctaa cttttgtatt ttttgtacag acatggtttc tccatgttgc 87660

ctgggctggt cttgaactcc tgaactcaag tgatcctccc acctcggcct cctaaagtgc 87720

tgtgattaga ggtatgagcc accgtgcccc acctgtactt gtgtgtttct gctcaaattt 87780

tattggggag cttttatggt atagcctgtg gtctctctgg agaatgtttg tgtctatgac 87840

aatgtatatt cttcagtatg ttatgttcca ttttggatgc ttgcatgggg actgtgtgtc 87900

cctgtactga aactcaggtg aacacttggc tctccgtcca ttgctgttct ctagaaatcc 87960

agcccagttc ccttggttaa agataaggta tgtctagcag gtattgctct ttctcttgca 88020

gacaaaactc aggaggattg ccccttgata aacaaggcta acctgctgat tctttgaagg 88080

aaggaactgg agattgtcct tttaggggtt tatattctgg attccagaaa acacacaaat 88140

agaaaacagg gcaaataaat gcatctttct tttcgagtcc attttaacct ggttaaggaa 88200

gattccaaca aaaaatccac ggtgccacag caagaagatg tcaggctgtg tccctctaca 88260

gggaagcgct ttctgttgtc tgaaagaaaa gaaagtgctt ccttttagag ggttaccgtt 88320

taagaaaagc aacgtttagg tggatgctga tcttggcaat aatacatttg cagagcatgc 88380

ttatcatcag agttggatga tggtggggtt ctgttttttt gttttttttt ttctgagaca 88440

gagtgtctat tgcccaggct ggagtgcagt ggcactttta acctagatct cttgggctca 88500

agtggtcctg ttttttggga taatctcaca ctgtgaccca gtccggagta cagtggcatg 88560

aacaccactc attgcatgct ttgcctccca ggttcaagtc attcttatga cccagcttcc 88620

agagcagctg ggattacagg catgagccac cacactggct aatttttgta tttttaatag 88680

agatggggtt caccatgttg gccaggctgg tctccaacta ctgacctcat gatccaccca 88740

cgtcggcctt ccaaagtgct gggattacag acgtgagcca ctgtgcccgg ccaacacaga 88800

gaatactctt atatcacttt ggggttgttt ctgtttgtgt attttttaaa tattttttga 88860

gacacagtct ggcgctatta cctaggttgg agtgcagtgt tgtgacctcg gctcacagca 88920

acctctacct cccagctgag cctcctgagt tagcatctgc aggcatgaac cacaacacct 88980

ggtaaccttt gtatttttgg acacatgagg tttcaccatg ttgcccaggc tagtcttgaa 89040

ctcctgagct caagtaatcc tcctgcctcc gactcctaaa gtgctgggat tacaggtgtg 89100

agccactgca ccccacctgt acttgtgtat ttctgttcaa attttattga ggagctttta 89160

cggcctagac agtagtctat cctggagaat gtttgtgtgt atgacaatgt atattctttt 89220

tttttatttt tattttttgg agacggagtc tcactctgtc acccaggctg gagtgcaatg 89280

gcatgatctc ggttcactga aacctccgcc tcctgggttc cagcgatcct cctgtctcag 89340

cctgctgagt agctaggatt acaggtgggc gcctggctaa tttagattcc attttggatg 89400

ctcccatcgg gactgtgtgt ccctatgttg gaactcaggt gaacgcttgg ctcaaaaatc 89460

cattgctgtt ctctagaaat cctgcctaat tctcttggtt aaagataagg tatatgtagc 89520

aggcattgct ttttctcttt ggggacaaaa ctcaggagga ttgcgccttg atgaacaagg 89580

ctaacctgct gagcctttga agcaaggaac tggagatggt ccttttaggg gtttatgttc 89640

tggattccag aaaacatgca aacagggcaa ataaatgcat ctttattttg tgtccatttt 89700

aacctggtca aggaaaattc caacagcaac atcaaaaaac cagtgttgga gcaagaatat 89760

gtcatgctgt ggccctccag agggaagcgc tttctgttgt ctgaaagaaa acaaagcgct 89820

cccctttaga ggtttacggt ttgagtaaag cagcgttgaa gttgatgctg atcttggtaa 89880

tacatttgca gagcgtgctt atcatcagac gtggacgatg gtggggttct gttttggttt 89940

tgtttttttc taagacaggg tctccgttgc ccagactgga gtgcggtggc acttctacct 90000

agatctcttg ggctcagatg gtcttctttt tatttcattt ttttaattgt ttgagatgga 90060

gtctcactct gtcacccagg ctggagtaca gtggcaggat ctccacaact acagcctccc 90120

aggttccaga cattctccca cctcagcctc ccgagtagct ggaattataa gcacccaccg 90180

ccatgccctg ctttttgtat ttttagacaa gaaagggttt caccatattg gccaggctca 90240

tctcaactct tgcccttaag taatcctcct ccctggcctc ccaaagtgct gagattacag 90300

gcgtgagcca ccgcgcccag cctcaagtgg tcctcttgag tcagcctctc aagtagttgg 90360

gactacgtgg ggcgtgccac catacttggc taagttttta atttttggta caattggggt 90420

ctctttttcc caggatagtc tcatctccac agcttatgtg gtcctacacg tgtgagccac 90480

ctcgtcttga tgacccattt caaagagagt tgacatggcc aggcattgtg gggcacacag 90540

tcccagccac tgcagaggcc agggtcggag ggacctttga tttccagact gtaccatgca 90600

ctgatcacac cttcgaatag ccactgcgct ccagcctggg ccaacatagc aagatcccat 90660

ctctttaaaa acagattaca tggcacctgg ttacagaggc tccatttggg ttggttattt 90720

gaaatgggtc tcccatcaat ttagtgtaat caatcgaaat catttccttc attaagagag 90780

taagtagccg gggcatggtg cctcatgcct gtaatcccag cactgtggga ggcaaaggca 90840

ggcggatcgc ctgaggtcag gagttcaaga ccagcctggc caatatggtg aaaccctatc 90900

actactaaaa atacaaaaat tagccgggtg tggtggcgtg aacctaggag gctgaggttg 90960

cagtgagctg agattgcacc attgcactcc aacctgggcg actgagctgg acttaaaaaa 91020

aaggtaagta agccttcaag tctatccaat cttaatttgt aatctcaaat ggatgtccat 91080

attaacaaaa ttttcttgtt tttcctgttt gtgtcttttt agtttaatct tcttgtcctt 91140

gacactgaaa tttttttctc tccaacaact tgccgtcgtt tctcggaata gagctgcttt 91200

ctctgcagga agggtgtagt tgattcaacc gccacccact cacgccagcc ccggtgagtt 91260

ctttgactgc agcttccctg attatgctat tttcttcggg taatgaaccc aaggtacctc 91320

gcatttaaag gctagttgct tggtaagctg tgaaatgttg gctgtggcta atatgagaaa 91380

caaactatgg tagacaagat gagtctcagt ggcagtgata attgtcacat gggacaaaac 91440

cacagatact tttcacaaaa accttgagga ccctagaagg gcctctctag taacaggtgg 91500

gatgcgccgc agctctcgtt gttgccgtag tgagcgatgc ctgttcgtcc agccctcaac 91560

accttttact ccgtggaagt tatgcctgca ctggtttaca gaacctccct tgacttgggt 91620

tgaggtagag ctgaagggaa cctcagtgtc ccttgcaggt gggatgttca ctacgtagca 91680

agagcacaaa ggtggagtgc gtgggctttg agtttctatt gggtaaatga agctgaaatg 91740

tagggcacat gagcacgtca tcaatgtaca attgatttaa tttttgtatt ttagaaacgg 91800

aggtctcgct atgccgccca gggtagaatg tgctctctcc ctctcctctt cctagtatct 91860

gggactacag gtgaatattt taatctatgc acaggcaaga agagggcaag gagggttctt 91920

ttgttctgtc tttgactttg tgaggcagtg tcactctgtc actcaagctg gagtgcaata 91980

atgtgatctc ggctcactgc aacctctgcc tcctgggttc aagcaatttt tttttttttt 92040

tttttttgag tcagtatctc actctgtccc tcagggtgga gtgcagcggc ccagtctctg 92100

ctcctgcaac ttctgactcc caagtaggta caattgcaga cacgtgtcac cacgcttggc 92160

aaaatttttt ttgtattttt agagctggga ttttaccttg ttggcgaggc tggtctggaa 92220

ctcctgacct caggtgactc gcccaccttg gcctcccaac aagttatgct gatcttggta 92280

ataacttcgc agagcgtgct tccatcagac atggaggatg gtggggtcct gttttgttgt 92340

tgttgttttt ctaagatagg gtctcttgcc caggctggag tgtgttggca cttccaacct 92400

aggtgtcttg ggctcaaatg gtcctctttt ttcggacagt cttgctctgt gacccatgct 92460

ggagtgtagt gacgtgatct cagctcactg caacctccac ctcccagttt taggcagttc 92520

tcctgcctca cattcctgag tagctgggat tataggtgcc tgacaccatg cctggctaat 92580

atttgcatgt tatttattca tgtatttatt tgttttgaga tggagtctca ctgtcactca 92640

ggccagaggg cagtggcagg acctcggctc actcaaacct ttgcctctca ggttcaagca 92700

agtcttctgc ctcaacctcc tgagtagctg gaatcacagg cacccgtcac cacgcccggt 92760

tacttttgta tttttttttt caccatgttg gccaggctgg tctgaaaccc ctgacttcat 92820

gtgagccgcc cgcctcagcc acccaaagtg gtgggattac aggcataagc cactgcgccc 92880

ggccaacaat gtatattttt caacatggga gattcagttt tgaatgctcc catcgggact 92940

gtgtgtccct gtgctggaac tcaagtgaac gcttggctca aaatccattg ctgttctcta 93000

gaaatccagc ccaattctct tggttaaatg taaggtatgt gtagtaggca atgctttttc 93060

tggagacaga actcaggagg attgtccctc gatgatctag gctaacctgc tgagactttg 93120

aagcaaggaa ctggagatgg tccttttagg gttttatgtt ctggattcca gaaaacatgc 93180

aaacagggcc aataaatgca tcctcatttt tgtgtccatt ttaacctggt gaaggaaaat 93240

tccaacaaaa aacccacagt gctggagcaa gaagagctca ggctgtgacc ctctagaggg 93300

aagcactttc tgttgcttga aagaagagaa agcgcttcct tttagaggat tactctttga 93360

gaaaaacaac attgaagtta atgctgatct tggagatcat gcttataatc agacttggat 93420

gatgttgggg ttttgtgggg tttttttggt tttttttcct aagacagggt gtctgttgcc 93480

caggctggag tatggtggcc cctccaacct agatctcttg ggttcaagtg gccctctttt 93540

ttgggacaga gtcttgctct gtgaccctgg ctggagctca gtatcaggaa ctttgctcac 93600

tgcaacctct gcctcccagg ttcaaatgat tctcctgcct cagcctcccg agcagctggg 93660

attacaggca tatgccacca cgcccagcta atttttgtat tattgttatg tatttatttt 93720

ttaacttatt cattttagag acagagtctc actctgtcgc ccaggctgga gtgcagtggt 93780

gcgatctcgg ctcactgcaa cctccaactc ccagctcagc ctcctgaata gctgtctgca 93840

ggcatacacc accacacctg gctaactttt ggtttttttg tacagatgtg gtttccccat 93900

gttgcccagg ttggtctcga actccttagc tcaggtgatc ttccaccttg gcctcctaaa 93960

gtgctaggat tagtggtgtg agctgttgtg ccccacctat actcgtctgt ttctgctcag 94020

attttattga ggagcgttta cagggtagcc tgttgtctac cctgcaggat gtttgtgcct 94080

acaacagtgt atattcttca acatcttaga ttccatttcg gatgccccca tgaggactgt 94140

gcgctcctgt actggaactc aagcgaccac ttggctcaaa atccattgct attctctaga 94200

aatccagccc aattctcttg gttaaagata aggtatgtgt agtaggcatt gccttttctc 94260

tttgggaaca aaactcagga ggattgcccg ttgatgaaca aggctaacct gctgattctt 94320

tgaagcaaag aactgcagat ggtcctttta gggatttatg ctctggattc cagaaaacat 94380

gcaaacaggg caaataaatg catctttatt tttgcgtcca ttttaacctg gtcaaggaag 94440

attcccacaa aaaatccaca gtgccagagc aagaagatct caggctgtcg tcctctagag 94500

ggaagcactt tctgttgtct gaaagaaaag aaagtgcttc cttttagagg gttaccgttt 94560

gagaaaagca acattgaagt tgatgctgat cttggtaata cattttcaga gcatgcttat 94620

catcagagat ggatgatggt gggcttctgt ttttgttttg attttttctt tttttttttt 94680

ttgagatgga gtttcgttct tgctgcccaa gcactagtat gtaggaatta ttctttatgt 94740

gagcatatag tacatggaac tttgtttttt tgagacaggg tctccaaaat atatacatgt 94800

tatcatataa tcttctatgt ataattatat tcacacaaaa tagtcttctg cacacagaga 94860

atactcttat atcactttgg ggttgttttt gtttgtgtat tttttttttt tttaattttt 94920

tttgagacac catctggctc tattgcccag gctggagtgc agtgttgtga cctcaactca 94980

ctgcagcctc tacctcccac ctcagcctcc tgaattagtg tctacaggca tgaaccacta 95040

cacctggcta tcttttgtat ttttacagat gaggttttac tatgttgggc aggctagttt 95100

tgaactcgag ctcatgtaat tttcccgcct tcgactccta aagtgctggg attagaggtg 95160

taagccactg cactccaccc gtacttctgt acaaatgtta ttgaggagct tttacggcct 95220

aggcagtgct ctatcctgga gaatgtttgt gtgtaagaca atgcatattc tttttttctt 95280

tcctgtcacc caggctggag tgcattggca cgatctcgtc tcactgcaac ctccgcctcc 95340

cgggttcaag caattctcct gcctcagcct cctgagtagc tgggattaca ggtgggcacc 95400

accacgcctg gcaaatttag attccatttt ggatgctccc atcatgactg tgtgtccctg 95460

tgctggaact caagtgacca tgtgactcaa tccattgctg ttctctagaa atccacctca 95520

attctcttgg ttaaaggtaa ggtatgtgta gtgggcattg ctttttctcc ttggagacaa 95580

aacttaggag gattcccctt gatgaacaag gctaatctgc tgagcctgga gattgtcctt 95640

ttagaggttt atgttctgga ttccagaaaa cggtcaaaca gcgaataaat gcatctttat 95700

atttgtgtcc atcttaacct ggtcaaggaa aatttcaaca aaaatcccag atggctggag 95760

cgagaagatc tcatgctgtg actctctgga gggaagcact ttctgttgtc tgaaagaaaa 95820

caaagcgctt ctctttagag tgttacggtt tgagaaaagc aacgttgaag ttgatgctga 95880

tcttggtaat acattcgcag agcatgctta tcaggcttgg atgacggcag ggttctgttt 95940

tggttttatt tttttccaag accagtctct gttgcccagg ctggagaggg aggggttgca 96000

ctccaaacta gatctcttgg ggtcatgtct tgctctatta ttattattaa ttcttatttt 96060

tgtgtgtgtg tgagacggag tctcgctctg tcaccaggtt ggagtccagt ggcggaatct 96120

cagctcactg caacctccga ctccgtggtt caagcgattc tcctgcctca gcctcctgag 96180

tagctgggat tacaggcacg caccgccatg ccgagctaat ttttgtattt ttagtagaga 96240

ccgggtttca ccacgttggc caggatggtc tcaaactcct gacctcatga tccacccgcc 96300

tcggcctccc aaagtgctgg gattacaggt gtgagcctcc gtgcccagcc tgttatttta 96360

ttttgtttta ttttatttat tttattttat tttgagatgg agactggctc tgtcgcccag 96420

gctggaatgc agtggcagga tctccactca ctgtaactcc cacctctgtc gcccaggctg 96480

gagtgcagtg gcaggatctc cactcaccgt ggagttccag tgattctttc accttagcct 96540

cgcaaatagc tggtattaca agcacccacc aacacaccct gccaattttt gtatttttag 96600

acaagaaagg gtttcgccat gttggccagg ctggtctcga ctcctgacct taagtgatcc 96660

acctccctgg cctcccaaag tgctgggatt acaggcgtga gccaccacgt ccagcctcaa 96720

gtggtcttct tgtcagcctc gcaagtagtt gggctcaggt tttaattttt ggtagagatg 96780

gggtctcttt tgcctgggat ggtctcagct gagcttaagt gatcctaaat gtgtgagcca 96840

ccttgtccca tttcaaagat aggtgacacg gccaggcatc ctgggacaca cagggtccca 96900

gttactgcaa aggctagggt gggagggtcc tttgatttta aggctatacc atgcactgat 96960

cacacctttg aatagccact gcactccggc ctgggccaca tagcaagatc ccatctcttt 97020

aaaaacgcag attacatgcc atctggttcc tgaggctcca tttgggttgg tcacttcaaa 97080

tacaggcctt tcatcagttt agtttaatca atccaaacca tgtttccttc attaggagag 97140

taagggccag gtgtgtaatc caagcacttt gggagactga agcaggcgga tcaggaggtc 97200

aggagttcga gaccagcctg accaatatgg tgaaaccccg tctctactaa aaatacaaag 97260

attaagtggg cgtggtggtg cgtgcttgta atcccagcta ctcaggaggc tgaggcagca 97320

gaatctcttc aacccaggag gcggaggttg cagtgagcca agatcgcgca ggtgcactcc 97380

agcctgggcg acagagcgag gctccatctc aaaacaaaac aaaacaaaaa atataggcct 97440

ccacatctat ccagtctatt ctgtaatccc aatggatgtc aatattaata caatgttctt 97500

attttttcta tttatttgta tctttttagt tgaatctcct tgtccctgac actaaatttt 97560

ttttattttt atttatttat tttttttgaa acagtcttgc tctgtcgccc aggctgcagt 97620

acagtggcgc aatctcggct tactgtagcc tctgtctccc gggttccagt gattctcctt 97680

ccccaactcc cgggtaactg ggattttcag gcacacacca ccaccccagc taattttttt 97740

gtacgtttag tagagacagg gtttcaccat attgggcagg ctggccttga actcctgaac 97800

taaggtgatc cgcccacctc ggcctcccaa agtgctggga ttagggataa ggctaacctg 97860

cttattcttt gtagcaagga actggagatg gtcattttag gggtttatgt tctggattcc 97920

agaaaacatg caaacagggc caataaatgc atcctcattt ttgtgtccat tttaacctgg 97980

gcaaggaaaa ttccaacaaa aaacccagag ttctggagca agaagatctc atgctgtgac 98040

cctacaaagg gaagcacttt ctcttgtcca aaggaaaaga aggcgcttcc ctttggagtg 98100

ttacggtttg agaaaagcag cgttgaagtt gatgcttatc tcggtaatac atttgtagag 98160

catgcttatc atgaggcttg gacgatggcg gggttctgtt ttggttttgc ttttttattc 98220

taagacagga tctctgttgc ccaggctgga gtgcggtggc acatccaacc taggtctttt 98280

ggattcaaat ggttttttta gcggacagag tctctctatc acccaggctg aagtgcagtg 98340

gtgtgatctc ggctcactgc aacctccacc tcccaggttc aagcgagtct cctgcctcag 98400

cttcccaagt aggtgagatt acaggtgccc atcaccacac ctggataata ttttcatttt 98460

ttaaaattca tttatttatg tttttgagat ggagtctcac tgttacccag gctggagggc 98520

cgaggcgtga tctctgctca gtgcaacctc tgcctcccgg attcaagcaa ttcttctacc 98580

tcagcctcct gagtagctag aattacggga gcccaccacc acacctggct actttttgtt 98640

ttttgttttt tgtttttttt ttagtagaga tgggatttca ccatgtttgc caggctggtc 98700

tcaaacccct gacctcaagt tagccaccgg cctctgcctc ccaaagtgct gggattacag 98760

gcatgagcca ccacgcctgg cctaattttt atattttcag cagaggcggg gtttcaccat 98820

gttggccagg ctggttgcga actcctgacc tcaggtgatc tgcccacctc agcctcccaa 98880

agtgctaaga ttacaggcgt gagccaccac acatggccat taacaccttt tagtccatgg 98940

aaattattct ggcactgatt tataaagcct catgtggggt caggtagagt caaaggggga 99000

cctcagtgtc ccttgcagat gggatgtgct cagcaagagc acggaggtgg agtgcatggg 99060

gtttgagttt tcactgggga aatgaaacca acatcttggg tgcatgacca ggtcatatat 99120

gcagtcatat atgcaatatg catggtggtg ggtgcctcta atcccagcta ctcaggaggc 99180

tgacgcagaa gaatcgcttg aacccaggag gcagaagttg ccatgagccg agatcccacc 99240

accgcatcca gcctgggcgg cagagtgaga ctccgtgtca aaaaaaagaa gatctcaggc 99300

agtgaccctc tagatggaag cactgtctgt tgtataaaag aaaagatcgt gcatcccttt 99360

agagtgttac tgtttgagac aagcaacgtt gaagatgctg ctgatcttgg taatacattt 99420

gcagagcgtg cttatcatca gacttgcatg atgtcggggt tctgtttgtg atttgaaatt 99480

tttccaagac aggctttcta ttgcccaggc gtgggtggat agcaccttcc accaagattt 99540

cttgggctta agtggtcctc tttttatttt ttgatttttt gagacacact cttgtttcgt 99600

tgggagtgca gtagcaggat ctctgctcac cggaaactcc acctctcggg ttccagtgat 99660

tctcccacct cagcctgccg aattgttggg aattcaggca tcaggcaccc accatcgtgt 99720

cctgcactct tgctgcccat cctggagagc actggcatga tctaggctca ctgcaacctc 99780

cgcctcctgg gttcgagtga tgcttctgcc tcagcctccc aggtagttgg gattacaggc 99840

acctactgcc ccacccagct aatttttatg tttttagtag agacgggttt caccatgttg 99900

gccaggctag tcttgaactc ccgctttcag gtgatccact agcctgtggt ctatcctgga 99960

ggatgtttgt gtgtgtgaca atgtatattc ttcaacatct tagactccat tttggatgct 100020

cccatcggaa ctatgtatcc ctgtgctgga actcaagtga acactcagct caaagtccat 100080

tgctgttctc tagaaatcca gcccagttct cttggttaaa tataaggtat gtgtagtagg 100140

cattgctttt tctctttaga ggcaaaactc aggagggttg ccccttgatg aacaaggcta 100200

acctgcagag cctttgaaga aaggaactgg agatggtcct tttaggggtt tatgttctgg 100260

attctagaaa acaggcaaac agggccaata aatgcatctt tattgttgtg gccattttaa 100320

cctggtcaag gaagatttca acaaaaaacc cagagtgctg gagcaagaag atcccatgct 100380

gtgaccctcc aaagggaagc gctttctgtt tgttttctct taaacaaagt gcctcccttt 100440

agagtgttac cgtttgggaa aagccacgtt gaagatgatg ctgatcttgg taatacattt 100500

gcagaccatg cttgtaatca gacttggatg atgttgggag tctgtttttt tgtttgtttg 100560

tttggttggt ttttttgttt gttaggtttt tgttttttgt tttggtgtgt gtgttttgtt 100620

ttgtgttttt ttttttttct aagacaaggt ctctgttgcc caggagagac tagagaagca 100680

ctttttaaga taggcctgtt gggctcagat ggtcctttta tgggatacag tcttgctctg 100740

tgacccagac tggagtgcag tggcgcgatc ttggctcaat gcaacctcca cctcccgggt 100800

tcgagtgatt ctcctgcctc agcttcctgg gtagctggga ttacagatgc ctgacatcac 100860

actcggctaa tatttgtatt tttctttttt attcattcat tcatttttga gatggagtgt 100920

tgctgtcacc cagcctggag ggcagtggca tgatcccggc tgactccaac ctctgcctct 100980

caggttcaag ccattctcct gcctcagtct cctgagtagg tggaattaca ggcacccacc 101040

actacaacca gctacttttt gattttgatt tttttttttt tttttttttt ttagtaaagg 101100

tggggtttca ccatgttggc caggctagtc tcaaacccct gacctcaagt gagccgctgg 101160

cctcggcctc ccaaagtgct gagaatacag gcgtgagcca ccgggcacag ccatcagcac 101220

cttttacttt atggaaattt ttctggcact ggtatagaac ctcacgtggg gtcaggtgga 101280

gttgagggga cctcagtgtc cctgcagatg ggatgagcaa gagcacggag gtggagtgca 101340

tggggcttca gtgtttattg gggaaatgaa gctgaaatct gggtgcatga ccaggagata 101400

aatgcatgag acggcggtct cactatgctg ccctggctga agtgggctta gatcctcctg 101460

cctctgcccc tccccagtcc ttggtagatg ggaccacatg tgaatattaa ccccccatgc 101520

acagacaaga agaaagtaag gactgttctt tggttcatac ctgaccccag ttaaacttgt 101580

attttagata aacaatgtat ttgagatgta cttgaacaac aaacgatttg ctgttcaggt 101640

gtgggcatct ttgttttgtt tttttttttc ctaattttaa atatgggact agtgtgcata 101700

tggtggctct tgtctgtaat tcctgcactt tgtgaggctg agacaggagg atcacctgag 101760

atcagttgtt cgagaccagc ctggccaaca tgaagccagg aggctgagct ttcagtgagc 101820

tgaaatcttc tgctgcactc caccctgggc aacagagtga gactcagtct caagaaagta 101880

aaaaataggc cgggcgcggt ggcgcatgcc tgtaatccca gcactttggg aggccgagac 101940

gggaggatca cctgaggtca ggagttcaag accaacaggg ccaacatggc aaagcctcgt 102000

ctctactaaa aatacaaaaa ttagctgggt gcaatggtgc acatctgtaa ttccagctac 102060

tccggaggct gaggcaggag aatcccttga acccaggagg aggagatttc agtgagctga 102120

aatcaagcca tggcattcca gcctgggcca gagagcaaca ttccgtctca aaaataaatg 102180

aataaataaa ataaataaaa ataaatagat aaatatggga catcataaat ttgggtgtca 102240

cctttgtgta gcagccagcg taatctctgt cattccaatt tttttttatg tgaactgcca 102300

aagcaagcac ttgtgtagga atttttcttc ctgtgagcat acaatatatg gaactttctt 102360

tgacacaggg tctcagaaat atatggtata tatttcgtta tatatatact tatatataat 102420

ataatcatat acacacaaaa tactgttcta aatacagaga atactctgat ataaccttgg 102480

gtattttttt ctttctttgt gtgtgtatgt tttttatttg tttggctggt ttttttttgt 102540

ttttgttttt tgttttttgt tttttgtttt tgtttttgtt ttgagacaga gtctcactct 102600

gtcgccgagg ctggagtgca gtgacaggat ctcgggtcac tgcaagctcc gcctaccggg 102660

ttcacgccat tctcctgcct cagcctcctg agtagctggg accacaggca cccaccacca 102720

cacctggcta attttttttt gtatttttag tagagacagg gtttcgccat gttagccagg 102780

atgatctcga tctcctgacc tcgtgatcca cccgcctcgg cctcccaaag tgctgggatt 102840

acaggcgtca gccaccgcgc ccggccagtt ggttattttt gagacaaggt ctggctctat 102900

cacccaggct ggagtgtagt ggtgcgatct tggctcactg caacctccac ctcccaccta 102960

aacctgctga attagtgtct acaggcatgc accaccacac ctggctcact tttgtgtttt 103020

tcatacacat gggtttacac catgttgctg gggctggtct caggaactcc tgagctcagg 103080

tgatcctccc gtcttggccc cctaatgtgc taggattaga ggtgtgagcc aacccacccc 103140

acccatactt acgtatttct gttcagattt tattgagttt ttatgaccaa gcctatggtc 103200

tatcctggag aatatttgtg tatgtgacaa tgtatattct tcaacatcgt agactcgatt 103260

ttgggtgctc ccatcgggac tgtgtgtacc tgtcctggaa ctcaagtgca cccttggctc 103320

ataatccatt gctcttctcc agaaatctta ccaattctcc tgcttaaata taagctacgt 103380

gtagtaggca ttgttttttc ttagcagata cgaaactcag gaagattgtc ctatgataaa 103440

caaggctaac ctgctgattg tttgaagcaa ggaactggag atggtccttt taggggttta 103500

tgttttggat tccagaaaac gtgtaaacag ggccaatacg tgcatcttta ttttgtgtcc 103560

agtttagcct ggtcaacaaa aatgtcaaca aaaaacccag agtgctggag taagaaggtc 103620

tcgggctgtg actctccaaa gggaagaatt ttctcttgtc taaaagaaaa gaacgcactt 103680

ccctttagag tgttaccgtg tgagaaaagc aatgctgaag ttgatgctga tcttgcaaat 103740

aactttgcag agcctgctta taatcagact ttgacaatgg tgggcgtctg tttttttttt 103800

tctaagacag gatctctgtt gcccaggctg gagtatggtg gcacttccaa cctaggtctc 103860

ttgggctcaa atggtcctct tttttgggac agagtcttgc tctgtgaccc aagctagacc 103920

acagtggcac gatcttggct cacggcaatc tctgcctccc agtttcaagt gattcccctg 103980

cctcaacttc ctgagtagct gggattatag gtgtctgaca ccatgactgg ctataatttt 104040

catttttctt atttatttat ttttgagtct cgctgtctct caggctggag tgcagtggca 104100

gcctttgcct cttgggttca agcaattttc ctgcctcagc ctcccaagta gttggaatta 104160

tatgtgtcac catcatgccc tttttgtttt tttttagtag aaaatgggtt tcaccacgtt 104220

ggccaggcta gtctcaagcc cctgacctca agtgatcctc ccgcctcaga ctcccaaagt 104280

gctgaaaatg caggcgtgag ccactgcacc gagccatcaa caccttctac tccatggaaa 104340

tgatactggc gctggtttat agaacctcac ttggggtcgg gcagatttaa aagggacctc 104400

aggctgggtg tggtggctga cgcctgtaat cccagcactt tgggaggccg aggcgggtgg 104460

atcacgaggt caggagatcg agaccatcct ggctaacatg gtgaaaccct gtctctacta 104520

aaaatacaaa aaattagcca ggcgtggtgg cgggtgcctg tagtcccagc tactcgggag 104580

gctgaggcag gagaatggtg tgaacccggg aggcggagct tgcagtgagc cgagatggca 104640

ccactgcact ccagcctggg cgacagagca agactctgtc tcaaaaaaat aaataaataa 104700

ataagggggt gggggacctc attgtccctt gcagatggga tgtgccctgc ttagcaagag 104760

cacggaggtg gagtgcatgg ctttgagttt tcactggggc acggaggtgg agtgcatggc 104820

tttgagtttt cactgggtaa atgcagctga actcttggct gcacgaccag gtcatatgtg 104880

caacgagaca ggggtctcac tatgctgccc aggctaaaat gggcttaggt ccttttgcct 104940

ccacctctcc ccagtcctta gtagctggga ctacatgtga atattaacca tgcacaggca 105000

agaggaaaga aaggaccgtt ctttggttca cacctagccg ccagtcaaat tagtatttta 105060

gatgaagact gcatttgaga catacttgaa caacaaatga tttgctgttt aggtgtgggc 105120

atctttcttt tttcctagtt ttaataatgg caccaggcag agtacagtgg cccacgcctg 105180

taattccagc actttgggag gctgagacag gaggatcacc tgaggtcggg aatttgattc 105240

cagcctggcc aacatggtgc agccccatct ctactaaaaa tacaaaatta gcccagcatg 105300

gtggtggtat atgcttgtaa tcccagctac tctggaggct gaggcaggag agtcgcttga 105360

agctggaagg ctgatctttc aatgagctga gatcacgcca ctgcactcaa gcctcggcaa 105420

aaagagtgag actccatctc aaaaacataa aaaataggcc agacacggtg gtccactcct 105480

gtaatcccag cactttgaga ggccaggaca ggcggatctc ctcaggtcag gagtccaaga 105540

ctagctgggc aaacaaggca aaacctcgtc tatactaaaa ttaggaaaaa tggctgggaa 105600

cagtggtgga cacctgtaat ctcagctact tgggaggcca cggcaagaga atctcttgaa 105660

cctgggaaaa ggagattgca gtgagacgaa atcacaccat tgtactccat cctgggtgac 105720

agagtgagat tctgtttcaa aaattaaata aataggccga gcacagtgac tcatgcctgt 105780

aatcccagaa ctttgggagg ccgaggtggc ggatcacctg aggtcgggag ttcaagacca 105840

gcctaaccaa cacgtagaaa ccccatctct actaaaaata caaaattagc cgggcatggt 105900

ggcgcatgcc tgtaatccca gctgctcggg aggttgaggc aagagaatca cttgaacccc 105960

agggggcgga ggttgtggtg agccgagatt gtgccattgt attccagcct gggcaacgag 106020

tgaaactctg tctctaaata aataaataaa actattaaat aaacattaaa cgttaaaaaa 106080

taaaaataga taaatatgag atcatcatga atttgagtgt cacctttgcg cagggcccat 106140

ggtaatcttt gtcattccaa ttttttcatg cgtgctgcca aagctagcac ttgtgtgtag 106200

gaagtattct tcctgtgagc atacaatata tggaagtttc ttttattatt attatttttt 106260

atgtttttgc gactgagttt tgctcttgtc gcataggctg aagtgcaatg gtgtgatctg 106320

ggctccctgc gatgtccacc tcttgggtta aagcgactct cctacctcag cctcctgagt 106380

agctgggatt acaggcatac acttccaggg ctagctaatt tgttgtattt tagtagacgg 106440

gatttctcca tgttggtcag gctggtctca aactcccgac ttcaggtgat ctgcccacct 106500

tggctttcca aagtactggg attgcagaca gccaccgcgg ctggccttca tatattctta 106560

tatatataat atcatgtaca cacaaaagac tcttttacac cctgagaata ctcttatatc 106620

actttgggca tttttttgtt ggtgtgtgca ttttttgttt gttcgttcgt ttgtttttga 106680

gacaagatct ggccctgtca cctatgctgg agtgcagtgg tgtgatctcg gctcccggca 106740

acctcaacct cccacctcag cctcctgaat tagtgtctac aggcatgcac ctctatacct 106800

ggctaacttt tgtattttta cagatgaggt tttaccatgt ttcccaggct ggtgtctaac 106860

tcctgagaat ataaggtata tgtagtaggc gttgcttttt ctctttgaag acaaaaccca 106920

ggacagttgt ccctcgatga acaaggctaa cctgctgaac gtttgaagca aggaactgga 106980

gatggtcttt gtatcggttt atgttctaga ttccagaaaa tatgcaaaca ggaccaataa 107040

atgcatcttt atttttgtgt ccattttgac caggtcaagg aaaatttcaa caagaaaccc 107100

agagtgccgg agcaagaaga tctcaagctg tgagtctaca aagggaagcc ctttctgttg 107160

tctaaaagaa aagaaagtgc ttctctttgg tgggttacgg tttgagaaaa gcaacgttga 107220

agttgatgct gatttcggta atacatttgc agagcatgct tatcacactt ggacgatggt 107280

ggggttctgt tttggttttg cttttttatt ctaaggctct gtgttgctca tgctggagtg 107340

cagtgacatg tccaatatat ctcttggctt caaatggtca gcctgggcaa cacagggaaa 107400

ttctgtctca aaaaataaac aaatgaataa aaatacaaaa taaagccggg catggtggct 107460

catgcctgta atcccagcac tttgggaggc cgatgcaggt agatcatctc aggtcaggag 107520

tttgagacca gtctgaccaa cgtggtgaaa atccgtctct actaaaaata caataacaac 107580

aaaaattagc ccagtgtggt ggtgggcacc tgtaatccca gctacttagg aggctgaggc 107640

aggagaattg cttgaacccg ggaggcagag gctgcagtga gccgagattg tgccacagca 107700

tccagccagg atgacagagt gagattccgt ctgaaaaaaa aaaaaaaaaa aaaaagctgg 107760

gcgctgtggc tcacgcctgt aatcccacca ctttgaaagg ccgaggcggg tggatcacga 107820

ggtcaggaga tcgagaccat cctggctaac acggtgaaac cccatctcta tttaaaaata 107880

caaaaaatta gcgaggcgtg gtggtgggtg cctgtagtcc cagctacccg ggaggctaag 107940

gcttgagaat ggcttgaacc caggaggcgg agcttgcagt gagccgagat cgtgccattg 108000

cactctagcc tcggcaacag agcaagactc cgtcaaaaag aaacaaaaac aaaaacaaaa 108060

aaaagaaccc tacagtactg gagcaagaag acctcatgct gtgaccctct agatggaagc 108120

actgtctgtt gtctaagaaa agatcgtgca tccctttaga gtgttactgt ttgagaaaag 108180

caacgttgaa gatgctgctg atcttggtaa tacatttgca gagcgtgctt atcatcagac 108240

ttgcatgatg ttggggttct ctttggtttg tttttttcca agacaggttc cctgttgccc 108300

aggcgggagt gggtggcacc tccaaccgag atctcttggg ctcaagtggt gatcttttta 108360

tttttgattt tttgagatgc gctctcattc cgtttcccag gctggagtgc agcggcagga 108420

tccctgttga ccggaaactc cgcctcccag cttccggcga ttctcccacc tcagcctgcc 108480

gaatagttgg gaatagagat gcccgccatc gtgtcctgct agttattatt tttattgttg 108540

ttgttgttgt tgagatggag tttcactctt gatgtccagg ctggagtgca gtgttgcaat 108600

ttaggctcac tgcaacctcc acctcctggg ttcaagcgat tctcctgccg cagcctcctg 108660

ggtagctggg attacaggtg cctactgccc cacccagcta atttttgtgt tttgaataga 108720

gactgggttt caccatgttg ggcaggctgg tctcaaactc ccgacctcag gtgatccacc 108780

agccttggcc tcctaaagtg ctgggattac aggcgtgagc caccgcccct aatctcacgt 108840

ggtcctctag agtcagccta gactctggtc tatcctggag gatgtttgtg tgtgagacaa 108900

tgtatattct tcaacatcag cccaattctc ttagctaaat gtaaggtatg agtagtaggt 108960

attccttttt ctctttgggg acaaaactca ggacgatcgc cccttgatga acaaggctaa 109020

cctgctgagc ctttgaagca aggaattgga gatggtcctt tcaggggttt atgttctgga 109080

ttccataaaa catgtaaaca gggccaataa atgcattttt atttttgtat ccgttttaac 109140

ctggtcaagg aaaattccaa caagaaaccc agagtgctgg agcaagaaga tcccatgctg 109200

tgaccctcta gaggaagcac tttctgtttg ttgtctgaga aaaaacaaag tgcttccctt 109260

tagagtgtta ccgtttggga aaagcagtgt tgaagttgat gctgatgttg gtaatatatt 109320

tgcatgctta ttatcagact tggatgatgt tggggttctg ttttgttttt gttttctaag 109380

acagggtctc tgtggcccag gctggagtac agaggcactt ccaacctagg tctcttgggc 109440

tcatatggtc ctttttggga cagtcttcct ctgtgaccca ggctggagtg cagtggcacg 109500

atcttggctc actgcaatct ccacctcgcg ggttcaagct attctcctgc ctcagcttcc 109560

tgaatagctg ggattgcagg ggcccgacat cacacttggc taatatttgt atttttcttt 109620

tttattcatt tatttatttt tgagatggag tctcgtttcc ccagctggag agcggtggca 109680

tgatcccgac tgactccaac ctctgcctct caggttcaag caattctcct gcctcagcct 109740

ctgaagtagc tgaaactaca ggtggccgcc accacgcctg gctacgtttt gaattttttt 109800

tttttttttt ttcagtagag atggggctta ccatgttggc caggctggtc tcaaacccct 109860

gacctcaagt gagccactca ccttggcctc cgaaaatgct gagaatacag gcatgagcca 109920

ccaccgcgcg cctcccaaag gcgtgagcca tggtgcgtgg ccatcaacac ctcttacttt 109980

atggaaattt ttctggcact ggtatagaac ctcacatggg gtcaggtgga gttgagggga 110040

cctcagtgtc cctgcagatg ggatgagcaa gagcacggag gtggagtgca tggggcttca 110100

gtgtgtattg gggaaatgaa gctgaaatct tgggtgcatg accaggagat aaatgcatga 110160

gacgggggtc tcactatgct gccctggcta aagtggtctt agatcctcct gcctctgccc 110220

ctcccaggcc ttgttagatg ggactacatg taaatattaa cccatgcaca ggcaggaaga 110280

aagtaaggac cattttttgg ttcgtcccgg ccctcagtta aacttgtgtt ttagataaac 110340

aatgtatttg agatgtgctt aaacaataaa tgatacgcta tttaggtgtg ggcatctttg 110400

tttcccccta attttaatga tgggactagt ccaggtatgg tggctcctgc ctataattcc 110460

agcactttgg gaggccgaga caggaggatc acctgaggtc agttgttcga gaccggcctg 110520

gccaacatgg tgaaaccccg tccatactaa aaatacaaaa atcagccagg cctggttgca 110580

cacacctgta atcccagcta ctcgggaggc tgaggcagga gactcgcttg aagccaggag 110640

gctgagcttt cagtgagctg agattgcgcc actgcattcc agcctgggca actgagtgag 110700

tctcagtgtc aacaaagtaa ataatagtct aggcacagcg gcacacgcct gtaatcccag 110760

cactttagga ggctaagacg ggaggatcac ctgaggtcag gagttcaaga ccaacggggc 110820

caacgtggca aagccttgtc tctactaaaa ctacaaaaat tagctgggtg cggtggtgca 110880

catctgtgat tccagctact cgggaggcca agacaggaga accacttgaa cccaggagga 110940

ggcgatttca gtgagctgta atgaagccat ggcattccag cctgggccac agagcaagat 111000

tccgtctcaa aaataaataa aacaaataaa aatacgtaga taaatacggg accgtcgtga 111060

atttgagtgt cacctttgtg gagcagccac ggtaatctct gtcattccaa ttttttttat 111120

gtgcactacc aaagcaagaa cttgtgtagg aattattctt cctgtgagta tgcgatgtat 111180

ggaactttcc ttgagacagg gtctcaaaaa tatatcatgt atattttata tattcttatc 111240

atatataata taatcatgta cacacaaaat actcttctaa atacagagaa tactctgata 111300

tcaccttgcg tattttttgt ttgcatgtgt ctgtgtttgg ttgttttttt tttgtcatat 111360

ttttgtatcc ctacttatat atttattttt aattttatta ttatacttta agttctaggg 111420

tacatgtgca caacgtgcag gttagttaca tatgtataca tgtgccatgc tggtgtgctg 111480

cacccattaa ctcatcattt agcattaggt atatctccta atgctatccc tccccgctac 111540

ccccacccaa caacagtccc cggtgtgtga tgttcccctt cctgtgtcca tgtgttctca 111600

ttgttcagtt cccacctatg agtgagaaca tgcggtgttt ggttttttgt ccttgcgata 111660

gtttgctgag aatgatggtt tccagtttca tccatgtccc tacaaagggg ttggttattt 111720

ttgagacaag gtctgactct atcgcccagg ctggagtgta gtggtgccat ctcggctcac 111780

tgcaacctcc acctcccacc taatcctgct gaattagtgt ctgcaggcat gcacaaccac 111840

atctggctca cttttgtatt ttttatacac atgggtttac accatgttgc aggcgctggt 111900

ctcaaactcc ctcccaagct caggtgatcc tctgtctcag cctcctaaag tgctgggatt 111960

agaggtgtga gccaccacac cccactggta tttgtgtgtt cctgttcaaa ttttattgag 112020

gagtttttat aaccaagcct gtggtccatc ctggaggatg tttgtgtgca tgacaatgta 112080

tattcttcaa catcgtagac tcgatgttgg atgctcccat cgggactgtg tgtccctgta 112140

ctggaactcg agtgaacact tggctctaag ttcattgctg ttttctagaa atccagccca 112200

attctcttgg ttaaatataa ggtgcacgta gtaggcattg ctttttcttt ctggagacaa 112260

aactcaggag gattgcccct tgatgaacaa agctaacctg ctgagacttt gaagcaagga 112320

actggagatg gtccttttaa gggtttatat tctggattcc agaaaatgtg caaacagggc 112380

caataaatgc atctttattt ttgtgtccat tttaacctag tcaaagaaaa ttacaacaaa 112440

aaatccacag tgctggagca agaagatctc atgatgtgac catctggagg taagaagcac 112500

tttgtgtttt gtgaaagaaa gtgcttcctt tcagagggtt actctttgag aaaagcagca 112560

ttgaagttga tgctgatctt ggtaatacat ttgaagagca tgcttatcat cagacttgga 112620

tgatcttggg gttttgtttt tttctaagac agggtgtctg tagcccaggc tcgagtgcgg 112680

tggcacttcc aacctagatc tcttgggctc aacaagtagc cctctttttc ggacagagtc 112740

tcgcgctgtg accctagctg gagtgcagtg gcaggaactt ggctcactgc cacctctgcc 112800

tcctgggttc aagcaattct cctgcctcag cctcccgagc agggttatag gcatgtgcca 112860

ccacacctgg ctaatttttg cattttatta tcgattgatt gattgattgt tgagacagag 112920

tctcactctg ttgcccaggc tgtagtgcag tggtgtgatc tcggctcact gcaacctcca 112980

cctcccacct cagccttctg aatacctgtc tacaggcatg caccaccaca cctcgccaac 113040

ttttgtgttt tttgtacaga tgaggtttca ccatgttgcc caggctggtc ttgaacccct 113100

gagctcaagt gatcctccca cctcggcctc ctaaagtgtg agtcactatg tcccgcccta 113160

cttgtgtgtt tctgcttaca ttttattgag gcgatttttt tttttttttt gagacggagt 113220

cttggtctgt tacctaggct ggagcgcagt ggcgcgatct tggctcactg caagctccgc 113280

ctcccaggtt catgccattc tcctgcctca gcctcctgag tagctgggac tacaggcgcc 113340

caccaccacg cccggctaat ttttttgtat ttttagtaga gatggagttt caccttgtta 113400

gccaggatgg tctagatctc ctgaccttgt gatcctcccg cctcggcctc ccaaagtgct 113460

gggattacag gcatgagcga ccgtattgag gcgcttttac aacatagcct gtggtctatc 113520

ctggggaatg tttgtgtgtg tgacaatgta tattcttaga catcttagat tccattttgg 113580

atgctcccat cggtactatg tgttcctata taggaactca agtgaacact tggcttaaaa 113640

tccattgctc ttgtctagaa atttagccca gttatcttgg ttaaagataa ggtgtgtctg 113700

gtaggcattg atttttcttt ccggagaaaa aacttaggag gatttcctct tgatgaacaa 113760

ggctagcctg ctgagccttt gaggcatggg acctggagat gatcctgtga ggggtttatg 113820

tttttggatt caggaaacac accagcagga caaataattg catctttatt tttgtgtcta 113880

ttttcaccag gctaaggaag attccaacaa catatccatg gtgctgaagt aagaagatct 113940

cgtgctgtga ccctctagag ggaagcactt tctgttgaaa gaaaagaaca tgcatccttt 114000

cagagggtta ctctttgaga agagcaacgt ttagtttgat gttgatcttg gtagtacatt 114060

tgcagagcat gcttttatca tcagacttgg ctgatggtgg tgttctgttt tttgttgttg 114120

ttttctacaa caggatgtct gttgcccagg ctagggtgtg gtggcactta caacctagat 114180

ctctagggct caaatggtcc tcttatttgg ggcagagtct tgctctgtga tccaggctgg 114240

agtgcagtgg caggaactct gctcactgca acctccgcct cccgggttca agtgattttc 114300

ctgccttggt tttctgagta ggtgggatta caggcgcaca ccaccacacc cagctagtat 114360

ttgtagtttt ctttattatt ctttttttta ttttaatttt tgagacgaag tctcgctgtt 114420

gcctacggtg gagtgcagtg gcgtgatctc ggctctctac aacgtctgcc tcccaggttc 114480

aagcaattct gctgcctcag cctcccgagt agctggaatc aacaggaaca caccaccatg 114540

cctggctact ttttggattt ttttttagta gagacggagt ttcaccatgt tggccaggct 114600

ggtctcaaac tcctgacctc atgtgagcca cccacctcgg ccttccaaag tgctggaatt 114660

acaggcatga accacgacgt ctggcctaat ttttgtattt ttagaagaga cagggtttca 114720

ccatgttggc caggctggtc tcagctcctg acctcaagtg atcctccccc tcggcctccc 114780

aaagtgctgg gattacaggc gtgagccacc tcgcccagcc tcaagtggtc ctcttgactc 114840

agcctcccat gtagttggga ctacatgggg cgtgccacca cacttggcta agtttttaat 114900

ttttcataca gatatgatct ctgttgctca ggaaggtctc aacacctgag ctcaaggatt 114960

ccacaggcgt gagccacatt ctcctgctga cccttttcaa agatagttga catggccagg 115020

catcatgggg cacacagtcc cagccaccgc agatcccggg gtgggagggt cctttgattt 115080

ccagactatt ccatgcactg atcacacctt gtttttttgt ttgtttgttt gtttgtttgt 115140

ttgtttgaca gagtctcagt ttgttgcctg gcgggagtgc aatgatgtga tctcgggtga 115200

ctgcaacctc tgcctccaag gttcaagtga ttctcctgcc tcaccctcca aagtagctgc 115260

ggttacaggc atgcaccacc acacccagct cagtttttgc atttggtaga gacaggtttt 115320

caccatgttg gtcaggctgg tctcgaactc ctgatttcag gtgatccacc tgcctcagcc 115380

tcccaaagag ctgggattat aggcgttagc ctatgcgccc ggtctgtatt tcctaatttt 115440

aaatatggga cccatggtga atttgggtgt cactgtgtac aggggacatg ggaatctctg 115500

tcattccaat ttttttaatg catgctgttc tgtgtaggaa ttattcttcc tgtgagcaaa 115560

caacatagag agctttcttt ttttgacacc gggtctcaaa aatatatcat atacatttta 115620

ttataatgtc tgatattgat aatataatca tacacacaca aaatactctt ctgtacacag 115680

agaatactga tatcactttg gggttttttt gtttgcgtgc gttttttggg attttggggt 115740

gttttttttt ttttttttga gacaacatct ggctctgttg cccatgctgg agtgcagtgg 115800

tgcaattggc tccctgcaac ctccacctcc cacctcccac ttcagcctcc taaagagctg 115860

aatacaggca tgcaccacca cacctgacta acttttgtat tttttgtaca cacagggttt 115920

accatgtttc ccaggctgga ctggaactcc agagctcaag tgatcctccc gacttggcct 115980

ccttaagtac tgcagttaga ggtatgagcc gccgagaccc accgtacttg tgtttttcta 116040

ttcaaatatt attgaggagc ttttatggcc taacctgtgg tctatcctgg aggttgtttg 116100

tgcgtacaat acatagtctt caatatcttt tttttttttt tcattttttg agagggagtt 116160

tcgctcctgt cacctaggct ggagtgcaat gacatgatct cagttcactg caacctccgc 116220

ctcctgggtt caagtgattc tcctgcctca gcctcctgag tagctgggat tacaggcacc 116280

cgccaccatg cctagctaag tttgtatttt cagtagagac gggcttctcc atgttggtca 116340

ggctgggctc agactcctga cctcaggtga tccgcccgcc tcggcctccc aaagtgccgg 116400

gattacaggc gtgagccacc gcgcccagtc atcatatatt cttatcatat atatagtagg 116460

atgtttgtgt atataacaat gtacattttc aacattgtag attccatttt ggatgctccc 116520

attgggactg tgtgtccctg tgctggaact caagtgaaca cttggctcaa aatccattgc 116580

tgttctctag aaatccagcc caattctctt ggttaaatat aaggtatgtg tagtaggcat 116640

tgctctttct ctttagagac aaagctcagg attgcccctt gatgaacaag gctaacctgc 116700

tgattctttg aagcaaggaa ctggagatgg tccttttagt ggtttatgtt ctggattcca 116760

gaaaacatgc aaacagggcc aatacatgca tctttacttt tgtgtccatt ttaacccggt 116820

gaaggaaaat tccaacaaaa aacccacaat gctggagcaa gatctcaggc tgtgaccctc 116880

tagagggaag cgctttctgt tggctaaaag aaaagaaagc gcttcccttc agagtgttaa 116940

cgctttgaga aaagcaacgt tgatcttggt aatacacttg cagagaatgc ttataatcaa 117000

ccatggaagg tggtggggtt ttgtgtttgt ttgttttgtt ttgttttcta agacagggtg 117060

tctgttgccc aggctggagt gtggcggccc ctccaaccta gatctcttgg gttcaagtgg 117120

ccctcttttt tgggacagag tcttgctctg tgaccctgcc ctggctggag tgtcctgtca 117180

ggatctccac tcactgcaac ctctgcctcc tggggtcaag ggattctcct gcctcagcct 117240

cccgagcagc tgggattaca ggtatgtgcc accacacctg gcttatttat ttatttattt 117300

atgagacaga gtcttgctgt gtcatccatg ctggagagca gtggtgcaat ctcggctctc 117360

tgcaacctcc acctcccacc tcagtctcct gaattagtat ctacaggcat gtaccaccat 117420

acctggctaa cttttgtatt tttacagatg aggttttacc atgtttcccg ggctggtgtc 117480

taactcctga gctcaagtga tcctcctgcc ttggcctacg taagtgctgc ggttagaggt 117540

gtgagccacc gcgccccacc tgcccttgtg tatctctgtt caaatgttac tgagaagcat 117600

ttacgggata atgcccggag gatgtttgtg tgtgtgacag tgtacattct tcaacatctt 117660

agattccatt ttggatgctc ccctcgggac tgtgtgtctc tgtcctggaa ttcaagtgaa 117720

cacctggctc tccatccatt gctgttctct agaaatccag cccaattctc ttggttaaat 117780

ataaggtagg tgtagtaggc attgcttttt ctctttgggg acaaaactca ggaggattgc 117840

cccttgatga acaaggctaa cctgctgatt ctttgaagca aagaactgga gatggtcctt 117900

ttaggggttt ttattcagga ttccagaaaa catgcaaaca gggccaaaaa ttgcatcttt 117960

atttttgtgt ccattttgac ctggtcaagg aaaatttcaa caagaaaccc agagtgccgg 118020

agcaagaaga tctcaagctg tgactgcaaa gggaagccct ttctgttgtc tgaaagaaga 118080

gaaagcgctt ccctttgctg gattacggtt tgagaaaaac aacactgaag ctgatgccga 118140

tcgtggtaat acatttgcag agcatgctta tcatcaggct tggacaatgc cggggttctg 118200

ttttcggttt tgctttttta ttctaagaca ggatctctgt tgcccaggct ggagtgcagt 118260

gacatgtcca acccaggtct cttgaattca aatggtcctt tttagggggc agagtctttc 118320

tctgtcaccc aggctgatat gcagtgaggc gatctcggct cactgcaacc tctgcctccc 118380

aggttcaagc aattctcctg cctcggcttc ccaagcagct gagattacag gcgttcatca 118440

ccatacctgg atgatatttg tattttttaa aatttattta tttatttatt tttgagacgg 118500

agtctcgctg tcacccagtc tggagggcag tggcacaatc tcggctcact gcaacctctg 118560

cctccccggt tcaagcaatt cttctgcctc agcctcctga gtagctggaa ttacagaagc 118620

acaccaccgc acctagttac tttttgtata ttttttttaa gtagagactg gatttcaccg 118680

tgttggctag gctggtctca aattcctgac ctcaagtgat ccacccacct ctgactccca 118740

aatttctggg atcacaggta tgcgccccca tgcctggcct aatttttgta tttttagcag 118800

agacgtggtt tcaccatgtt ggccaggctg aactcagctc ctgaccttag gtgatctgcc 118860

cgcctgggcc tcccaaagtg ctgagaatac aggcgtgagc caccgtgcct ggccattaac 118920

accttttggt ccacggaaat tattctggca ctggtttata gaacctcgct tggggtcagg 118980

tagagttgaa ggggacccca atgtccttgc aggtgggatg tgcactgctt agcaagaaca 119040

cggaggtgga atgcatgggg tttgagtttt tattggagaa ataaagccaa aatcttgggt 119100

gcatgaccag gtcgtatatg caacaatacg ggcgtctcac tatgccgccc aggctaatgt 119160

gggcttagat cctcctgcct ctgcccctcc ccagtcctta gtagctggga ctacatgtga 119220

atattaactc acgtacagga agaggaaagt aaggactgtt ctttgattca cgtcccaccc 119280

ccagttaaat ttgtatttta gataaacaat gtatttgaga tgtacttgaa caacaaatga 119340

tttgctgttt agatgtgagc atcttttttt gctcctgatt ttaaacatag gacaaggtca 119400

ggtatggtgg ctcacgcctg taattccatc attttggtag gcagagacag gaggatcacc 119460

tgaggtcagc tgcttgagac cagcctggcc aacatggtga aaccccatct gtactaaaaa 119520

tacaaaaatt agccatgcgt ggttgtgctg cacgcctgta atcccagcta ctcaggagga 119580

tgaggcagga gaatcgcttg aagccgggag gcagaggttg cagtgagctg agatcgcgcc 119640

agtgaactac agcctgggca aaagaataag attcaatctc aaaaaaataa aaaataggcc 119700

aggcacggtg gcacacacct gtaatgctag cacttttgga ggtcgagacg ggggcatcac 119760

ctcaggtcag gagttgaaga ccagctgcgg catcacggca aagcctcatc tctacgaaaa 119820

atacaaaaat tagccaggtg cagtggcagg tgcctgtaat accagctact cgggagacca 119880

aggcaggaga atcgttccaa ctgggcggca gaggttgcag tgagccaaga tcgtgccact 119940

gcactgcagc ctgtcgacag agtaagactt ggtctaaaaa aaaaaaataa ataaataaat 120000

aaaagataaa tatggaacca tcatgaattt gggtgtcacc tctgtgtggg ggccagggta 120060

atctctgtca atccaatttt tttttatgtg caccagcaaa gcaagcactt atgtttaggt 120120

attattcttc ctgtgagcat acaatatgta gatctttatt ttctttgaga catggtctca 120180

aaaatgtatg atgtggctgg gcacagtggc tcaagcctgt aatcccagca ctttgggagg 120240

ctgaggcgtg tggatcaccc gaggtcagga gttcaagacc agcctggcca acatggtgaa 120300

aatccgtctc tacgataaat acatatatat atatatgtat atatgccggg catggtggca 120360

ggtacgggta atcccagcta cttgggagcc tgaggcagga gaatcgcttg aacctgggag 120420

gtagaggtta ctcatatcac tatgagattg ttttttgttc ttgtgtgtgt ggtttttgtt 120480

tgtttttgag acaaggtctg gctctgtcac ccaggcaaaa gtgtagtagt gcgatctcgg 120540

cttactgtaa cctccatctc ccacctatgc gtcctgaatt agtgtgtcta caggcatgca 120600

ccaccacagt gggctacctt ttttttttga gagggaattt tgctcttgtt gcacaggctg 120660

gagtacaatg gcaagatttc agatcactgc aacctccgcc ttccgggttc atgtgattct 120720

cctgcctcag cctcccaagt agctgcgatt acaggcatgc accatcacac ttggctaatt 120780

ttgtattttg agtagataca ggtttcccca tgcttgtcag gctggtctcc aactcctgac 120840

ctctagtgat ccacctgcct cggactccca aattgctggg attacaggca tgagctacca 120900

cacccagcca cctggctaac ttttgtagtt tttctacaga ctacaaaatg gtttcaccat 120960

tttacctgag actacaaaat ggtttcacca ttttacctga gactacaaaa tggtttcacc 121020

attttacctg agctggtctc caacacctga gctaaagtga tcctcctgcc tcggcctcat 121080

aaaatgttgg gattagaggt gtgagccacc gtgcctcacc cgtacttgtg tatttcttta 121140

tttttattga tttatttatt ttgagacaga gtttcactct tgttgcccaa gctggagtgc 121200

aatggcacaa tctcggctca atgcaacctc cttctcttag gttcaagcga ttctcctgcc 121260

tcagcttccc gggtagctgg gattaaaggc gtccaccacc acacccagct aatttttgtg 121320

tttttagtag agacagggtt tcagcatgtt ggccagactg gtctcaaact cctaacctca 121380

ggtaatccac ctgcctcagc cttccaaact gctgggatta caggcatgag ccactgcgcc 121440

tagcctcaag tggtcctctt aagtcagcct aaactgtggt ctatcctgga ggatgcttgt 121500

gtgtgtgaca atttatattc ttcaacgtct tagattctat tttggaggct cccaagggga 121560

ttgtgtgtcc ctgtgctgga actcaagtga acacttggct gtacatccat tgctgttctc 121620

tagaaattca gcccaattct cttggttaaa gataaggtat gtgtagtagg cattgctttt 121680

tctttccaga gacaaaactc aggaggattg ccccttgatg agcaaggcta acctgctgat 121740

tctttgaagc aaagaaccag agatggtcct tttaggggtt tatgttctgg attccagaaa 121800

acaggcaaac aggaccaata aatatatttt tatttttcta tccactttaa cctggtcaag 121860

gaaaattcca acaagaaacc cagagtgctg gagtgagaag atcccatgct gtgaccctct 121920

agagggaagc actttctgtt gtctgaaaga aaccaaagcg cttccctttg gagcgttacg 121980

gtttgagaaa ctcaatgttg aagttgatgc tgacttcagt aatacatttg tagaggatgc 122040

ttatcatcag acttggatga tatcggggtt ctgtgctttt tttttttttt ttctcctaag 122100

acaaaatctg tattttccag gctggagtcc ataagcactt ccaacctagg tctcttgcgc 122160

tcagatagtc ctcttttttg ggacagagtc ttgctctgtg acccaggctg gagtgcagtg 122220

gcgtgatctc ggctcactgc aacctctgtc tcctgggctc aagcaattct ccttcctgag 122280

cttcctgagg agctgggatt acaggcgcct gacatcatcc ttggctaaca tttgtatttt 122340

tcttttttat tcatgtattt atttactttt cagatggagt ctcgctgtcg accaggctag 122400

agggcagtgg cacgatcccg gctgactaca acctgtacct ctcaggttca agcaattctc 122460

ctgcctcagc ctctggagta gctggaatta caggaacaca ccaccaggcc cggctacttt 122520

ttgtattatt attattatta ttattttttt tttttttcac taaagagggg gtttcaccat 122580

gttgtccaga gaatacaagc atgagccact gcgcagccat caacaccttt tactgcatgg 122640

aaattattct ggcactggta tagaacctca catggggtca ggtggagttg aggggacctc 122700

agtgtccctg cagatgggat gagcaagagc acggaggtgg agtgcatggg gcttcagtgt 122760

ttattgggga aatgaagctg aaatcttggg tgcatgacca ggaaataaat gcatgagaca 122820

ggggtctcac tgtgccgccc tggctaaagt ggtcttagat cctcctgcct ctgcccctcc 122880

ccagtcctag gtagatggga ctacatgtga atattaaccc atgcacagac aagaagaaag 122940

taaggactgt tattttgttc gtacccagcc cccagttaaa tttatcatat atataatata 123000

tcatatatat aacataatca tatacacaca aaatgctgat ttgaatacag agaatactct 123060

gatatcactt tgggtatttt ttggtttgtt tctgtgtgtg tgtgttttat ttgtttggtt 123120

ggttattttt gagacaaggt ctggctctat cacccaggct ggagtgtagt ggtgcgttct 123180

cagctcactg caacctccgc cttccaccta agcctgctgc attagtgtct acaggcatgc 123240

accaccacac ccggctcact tttgtatttt ttgtacacat ggggttacac catgttgccg 123300

gggctggtct cgaactcctg acctcaggtg attctcccgt cttggcctcc taaagtgcta 123360

gaattagagg tgtgagccac cgtgccccac ctgtacttgt gtatttatgt taaaattgtg 123420

ttaaggagtt tttatgacca agcctgtgat ctattctgga gaatatttgt gcgtatgaca 123480

atgtatattc ttcaacatcg tagattccat tttggatgct cccactggga ctgtgtgcct 123540

atactggaac tcaagtgaac acttggctca aaatccattg ctgttctcta gaaatccagc 123600

ccaattctct tggttaaata taaggtatgt gtagtaggca ttgctttttc tttctagaga 123660

caaaactcag gaggattgcc ccttgatgaa caaggtaacc tgctgattct ttgaaacaag 123720

aaactggaga tggttccttt aggggtttat gttctggatt ccaggaaaca tgcaaacagg 123780

gcccacaaat gcatctttat gtttgtgtcc attttaacct ggtcgaagaa cattccaaca 123840

aaaaatccac catgcaccga gcaagaatat ctcaggctgt gaccatctgg aggtaagaag 123900

cactttctgt tttgtgaaag aaaagaaagt gcttcctttc agagggttac tctttgagaa 123960

aagcaacgtt gaagttgatg ctgatcttgg taatacattt ggagagcatg cttatcatca 124020

gacgtggatg acggttgggt tttgtttttg ttttattttt tatcttaggc agggtctctg 124080

ttgcccaggc tggagtgtgt tggcacttcc aacctagatc tcttggtctt aaatggtcct 124140

ctttttgggg ggcggagtct cactctgacc caggctggag tgcaatggca ggaactcccc 124200

tcactgcaac ctccacctcc cgggttcaag cgattctcct gcctcagttt tccgagtcgc 124260

taggattaca ggctcactcc tccacacctg gctaatattt gtagttttct ttattattca 124320

tttatttatg tgtttttgag aaggagtctc gctgtcgccc aggctggagt gcagctgcac 124380

aatctcagct cactgcaact tctgccactg gagtaactgg gatttcaggt gcgtgccacc 124440

acacttggcc aattttttat gtgtgtattt ttagacccgg ggttttccca tgttggccag 124500

gctggtctgg aactcctgac ctcaggtgac ccatgcacct cggcctccca gagtgctggg 124560

attacaggca tcagccaccg cacccggccc tgtctctgac ttttgcgaac tgaggaagta 124620

accttttgaa atttactctg aagtgtgaat gattttagca tattttcaac caccaacatt 124680

ctagttcagg agattttcat caccccaaag aggcttattc ccatttgcag tcagttccca 124740

ccccactctt ccacccagac cgtgacaaac actttgcatc tctctagctc tggatctgcc 124800

tcttgtgggc atgtcacata gatcagtttt gaataggtgg cctgttgtga ccactgtaat 124860

tgaggataat gggttggtcc tggttgtctg cagtgtgaat cttaccactg aagggtggtc 124920

cctggatgga agcaggaggc tgggagaact gggcggaaca tcctttggga atggagtcgg 124980

gcgggcagac cctgatgcct gggaagctca caagggtgga agaccacatc ttcctccctg 125040

agaactgcaa ggtgaccctc ctggggtact ggaaggagtg aaggcctctg gactgggaac 125100

accagggcat tgcaccggtg caggcaggat gagccaaggg gaacggagag ccaggcatcg 125160

ctaactggcg acaatttggg tttgatctgg acggagtctg tgtcttctgg tagagagaac 125220

ccctgggatt ttcgctctgc tcctggctgt ctttcagtca tggaatctga tgacaaagac 125280

tcccgcccag agccaagaca tttggtttct ggaccccagt ggtcctttct gcctggactt 125340

gggatctttt ggggaagttt gggatctggc agggcatctg cataatccat agaaatccct 125400

gagagtccct tccctttgct gacatctcca tgttcctacc tattagcttc caaaggagac 125460

tcctatctga attgccgaag gcagcttccc aggccaggga tacccagtta aatttgtatt 125520

tcagatcaat agtatatttc agatgtactt gaacaagaaa tgatttgctg tttaggtgtg 125580

ggcatttgta tttccccctt tttttttttt tttttttttt tttgacagag ttttgctctt 125640

cttacccagg ctggaatgca atggcacaat cttgactcac tgcaacctcc acctcccagg 125700

ttcaagcgat tctcctgcct cagcagcccg agtagctggg attacaggca tgcgccacca 125760

cacccggctc agtttttgca tttagtggag atgggggttc accatgttag ccaggctggt 125820

ctcaaactcc tgacctcaag tgatttccct gcatcggcct cccaaagtgc tgggattata 125880

ggcatgagcc tccacgccta gcctcaagtg gtcctcttga gtcaacctag actacagtct 125940

atgctagaga atgtttgtgt gtgtgacaat gtatgttctt gaacatcata gattccactt 126000

tggatgctcc tgtcgggact gtgtgtccct gtgctggaac tcaagtgaga gttggctgaa 126060

aatccattgc tgttctctag aaatccagcc caagtctctt ggttagagat aaggtatgtc 126120

tagtgggcat tgctttttct ttttggggac aaaactcaga aggattgccc cttgatgaac 126180

aaggctaacc tgctgattct ttgaaggaag gaactggaga tggtctttta ggggtttata 126240

ttctggattc cagaaaactt gcaaacaggg ccaatacatg catctttata tttgtgtccg 126300

ttttgaactg gttaaggaaa atttcaacaa gaaacccaga gtgccggagc aagaagatct 126360

caagctgtgg gtctgcaaag ggaagccctt tctgttgtct aaaagaagag aaagcgcttc 126420

cctttgctgg attacggttt gagaaagcga cgttgaagtt gatgcatttg cagagcatgc 126480

ttatcatcag gcttggacaa tggcggggtt ctgttttggt tttgcttttt cattctaaga 126540

caggatctct gttgcccagg ctggagtgcg gtggcagtgg catgtccaac ctagccctct 126600

tggattcaaa tggtcctttt ttgcaagcaa agtctctctc tctcacccag gctggagtgc 126660

agtgatgtga tctcggctcg ccgcaacccc cgcctcctgg gttcaagcga ttctcctgct 126720

tcagcttccc aagtagctga gattacaggc acccatcacc acacctggat aatatttgta 126780

tttttaaaaa ttcatttatt tatttatgag acggagtctc gctgtcaccc atgctggagg 126840

gcagtgacac gatctcggct cactgcaatc tctgcctcct gggttcaagc aattcttcct 126900

cagcctcctg actagctgga attacaggag cacaccaccg cacctggcta ctttttgtat 126960

ttttttttta gtagaaatgg gatttcacca tgttggttag actggtctca aactcctgac 127020

ctcaagtcat ccgcccacct ctgactcttt ctgggattgc aggtatgcgc ccccacgcct 127080

ggcctaattt ttgaatgttt agcagagatg gggtttcacc atgttggcca ggctgaactc 127140

agctcctgac cttaggtgat ctgcctgcct cggcctccca aagtgctgag aatacaggcg 127200

tgagccaccg cgcctggcca tcaacacctt ttaccccaca gaaattattc tggcactggt 127260

ttatggaact tcacttgggg tcaggtagag gtgaagggga ccccaatgtc ccttgcagat 127320

gggatgtgta ctgctcagca agagcatgga ggtggagtgc atggggtttg agttttcatt 127380

ggggaaatga agctgaaatc ttgggtgcat gaccaggtcg tatatgcaac gagacgggcg 127440

tctcactatg ccgcccaagc taaagtgggc ttagatcctc ctgcctctgg ccctccccag 127500

tccttagtag ctgggactac atgtgaatat taactcatgt acaggaagag gaaagtaagg 127560

actgttcttt gattcatgcc ccacccccac ttaaatttct atttcagata aacaatgtat 127620

ttgagatgta cttgaacaac aaatgatttg ctgtttaggt gtgggcatct ttttttgttc 127680

ctaattttaa gtataggaca aggctgggta ccacggctca cacctgtgat tccatcattt 127740

tgggaggcca agacaggagg atcacctgag gtcagttgtt tgagaccagc ctggccaaca 127800

tggtgaatcc ccatctctac taaaaataca aaacttagcc cagtgtggtg gtggtacacg 127860

cctgtaatcc cagctactca ggaggctgag gcaggagaat cacaggaagc cgggaagctg 127920

agctctcagt gagctgagat cgcaccactg aactccagcc taggcaacag tgagactcaa 127980

tctcaaaaaa ataaaaaata ggccgggtgc ggtggcacac gcttgtaatc ccagcacttt 128040

gggaggtcga gacgggtgga tcacctcagg tcagagttca agagcagccc cgccatcaag 128100

acaaaacctc ctctctacta aaaatgcaaa atatagctag gcgtggtggt acacacctgt 128160

agtctcagct acgtgggagg cggcagcagg agaatctctt gaacccgtga gaaggagatt 128220

gcagtgagct aaaatcacac cattgcactg cagcctgggc gactccatct caaaaaataa 128280

atacataaat aaataaatac ataaatacat aaaaaataac ccagagtact ggagcaagaa 128340

gatctcaggc agtgaccctc tagatggaag cactgtctgt tgtctaagaa aagatcgtgc 128400

atccttttag agtgttactg tttgagaaaa tcaacgttga agatgctgct gatcttggta 128460

acacatttgc agagcgtgct tatcatcaga cgtgcatgat gttggggttc tgtttgtttt 128520

gttttttttt tccaagacag tgtctctgtt gcccagacta gagtgggtgg cacctccaac 128580

cgagatctct tgggctcaag tggtcctctt tttatttttt gattttttga gacccactct 128640

ggtttcactg ggagtgcagt agcagaatct ctgttgaccg gaaactccac ctctcaggtt 128700

ccagtgtttc taccacctca gcctgccgaa tcattgggaa ttcagttgcc tgccatcagg 128760

tcttgctaat tattattgtt tttttgtttt ttttttttga ggcggagttt cgctcttgtt 128820

gcccaggctg gagggcaata gcgcgatctc ggctcaccgc aacctcggcc tcccgggttc 128880

aagtgattct cctgcctcag cctcccagat agctggaatt acaggtgcct accaccccac 128940

ccagctaatt tttgtgtttt cagtagagat ggggtttcac catgttgggc aggctggtct 129000

caaactcctg acctcaggtg atccaccagc cttggcctcc caaagtactg ggattacagc 129060

tgtgagccat cacccctaat ctcaaatggt cctgtagagt cagcctagac tgtggtctat 129120

cttggaggat gtttgtgtgt gagacaatgt atattcttca acattgtaga ttccattttg 129180

gatgcttcca tcgggactgt gtgcccctgt cctggaactc agctgaacac ttggctcaaa 129240

atccattgct gttctctgga aattctgccc taatctcttg gttaaatata aggtatgtgt 129300

agtaggcttt gctttttctc tttggggaca aaactcagga gggttgcccc ttggtgaaca 129360

aggctaacct gctgagcctt tgaagcaagg aactggagat ggtcctttta ggggtttatg 129420

ttctggattc cagaaaatat gcaaacaggg ccaataaatg catctttatt ttgtgtgtat 129480

tttaacctgg tcaaggaaaa ttccaacaag aaacccagag tgctggagca agaagatccc 129540

atgctgtgac cctctagagg aagcactttc tgtttgttgt ctgagaaaaa acaaagtgct 129600

tccctttaga gttactgttt gggaaaagca gtgttgaagt tgatgctgat gttggtaata 129660

tatttgcatg cttatcatca gacttggatg atgttggggt tctgggtttt ttttgttttc 129720

taagacaggg tctctgtggc ccaggctgga gtacagaggc acttccaacc taggtctctt 129780

gggctcaaat ggtccttttt gggacagagt cttgctctgt gactcaggct ggagtgcagt 129840

ggtgcaatct cagctcactg caccctccac ctcctgggtt caagtgattc tcctgcctca 129900

gcttcctggg attacaggca cctgacatca tgcccagcta atatttgtat ttttcttttt 129960

tattcattca tttatttatt ttttaggtgg agtcttgctg ttgccaagtc tggagggcca 130020

tggcacgatc ccggctgact ccaacctctg tctctcaggt tcaagcaatt ctcctgcctc 130080

agcctcctga gtagctggaa ttacaggcgc ttaccaccac gcctgactgc ttttcgaatg 130140

tgtgcgtatg tgtgtgtatg tgtgttagag agaaagagag agagagagat ggagtttcac 130200

tcttgttgcc caggctggag ggcaatgaca tgatctcggc ccatacaacc tcagcctcct 130260

gggttcaagt gattctcctg cctcagcctc ctgagtagct aggattacag gcatgtgcca 130320

ccacacctgg ctaattttgt atttttagta gatatggggc ttccccatgt tggtcaggct 130380

ggtctcgaac tcctgacctc aaatgattca cctgcctcag tctcccaaag tgctgggatt 130440

acaggtgtga gccaacatgc ccggcctgaa tttttttttt tttttttttt ttttcaagta 130500

aagacggggt ttcaccatgt tgaccaggct ggtctcaaac ctctgaactc acgtgagctg 130560

ctcgcctcgg cctcccaaag gcatgagcca tggtgcgtgg ccatcaacac ctcttacttt 130620

atcgaaattt ttctggcact ggtatagaac ctcacgtggg gtcaggtgga gttgaaggga 130680

cctcagtgtc cctgcagatg ggatgagcaa gagcacggag gtggagtgca tggggcttca 130740

gtgtttattg gggaaatgaa gctgaaatct tgggtgcgtg accaggaaat aaatgcatga 130800

gacgggggtc tcactatgcc accctggcta aagtgggctt agatcctcct gcctctgccc 130860

cccgcagtcc ttgctagatg ggactacatg tgaatattaa cccatgcaca gacaagaaga 130920

atgtaagaac cattatttgg ttcacaccgg gccctcagtt aaacttgtat tttagacaaa 130980

caatgtatct gagatgtcct tgaacaacaa atgatttgct gttcaggtgt gggcatcttt 131040

gttttttccc taattttaat gatgggacta gtctgggtac ggtggctccc gcctgtaatt 131100

ccagcacttt gggaggctga gataggagga tcacctgagg tcagttgttt gagaccagcc 131160

tcgccaacat gatgaaacct cgtctgaact aaaaatatag atattagcca gacggggtgg 131220

tggtgcaccc ctgcaatccc agctactcag gaggctgagg caggagaatc acttgaagcc 131280

aggaggctga gctttcagtg agctgagatt gagccactgc actccagcct gggcaacaga 131340

gtgagactca gtctcaaaaa agtaaaaaat aggctgggtg tgggccgggc gcggtggctc 131400

acgcctgtaa tcccagcact ttgggaggcc gaggtgggtg gatcacaagg tcaggagact 131460

gagaccatcc tggctaacac ggtgaaaccc tgtctctagt aaaaatacaa aaaaaaaaaa 131520

aaaattagcc agacgtgatg gtgtgcgcct gtactcctag ctactcagaa ggctgaggca 131580

ggagaatggc ctgagcccgg gaggcggagc ttgcagttag ctgagatcgc accactgcac 131640

tccagcctgg gtgacagagc aagactccgt ctcaaaaaaa aaaaaaaaaa aaaaaaggcg 131700

gggtgtggta gtgcacatct gtaatcccag catttcggga ggccaagaca ggaattacct 131760

gaggtcagga gttcaagacc aacagggcca acatggcaaa acctcgtctc tactaaaaat 131820

acaaaaatta tctgggtgca gtggtacaca tctgtaattc cggctaccca gaggctgagg 131880

caggagaata gcttgaaccc aggaggagga gatttcagtg agctgaaatc acaccattgc 131940

acttcagcct gggccacaga gcaagattgt gtctcaaaaa taaataaata ataaaaatga 132000

cataaaatca ataaataaaa ataaatacat aaatacagga acatcattaa tttggatgtc 132060

acctttgtgc aggggccagg gtaatctctg tcattccaat tttttttatg tgcactgcca 132120

aagcaagcac tcatgtagga attatccttc ctgtgagcat ataatatatg gaacattctt 132180

tgagacaggg tctcaaaaat atgatgtgta tttcatcata tatataatat aatcgtatac 132240

acacaaaata ctgtttaaat acagagaata ccctgatatc accttgggta tttttctttg 132300

tttctgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg tgtgtttgtt tggttatttc 132360

tgagacaagg tctggctcta tcgcccaggc tggagtgtag tggtgtgatc tcaactcact 132420

gcaacctccg ccccccacct aaacctgctg aattagtgtc tacaggcatg caccagcaca 132480

cctggctaac ttttgtattt tttgtacata tggggttaca ccatgttgcc ggggctggcc 132540

tctaactcct gagctcaggt gatcctcccg cctcagcccc taaagtgctg ggattagagg 132600

tgtgagccac cgcaccccag ccatacttgt gtatttctat tcaaatttta ttgaggagtt 132660

tttaccgccc agcctgtggt ctatcctgga gaatgtttgt gtgtgcgaca atgtatattc 132720

ttcaccatcg tagattccgt tttggatgct cccatcggga ctgtgtgtcc ctgtgctgga 132780

actcgagtga acacttggct caaaatccac tgctgttctc tagagatctg tagggtccag 132840

ccctactggg cctgtgggtt tttctcttcg tgagcagata ggagagattg tagaaataaa 132900

gacacaagac aaagagagag aagaaaaggc agctgggcct gggggaccac taccaccaag 132960

acgtggagac cggtagtggc cccgaatgcg tggccgcgcc gttatttatt gtatacgagg 133020

caaaagggca gggtaagaag tgtgagtctt ctctaatgat aggtaagatg acgcgagtca 133080

cgtgtccacc ggacaggggg ccccttccct atttggtagc ttaggcggag agagattggg 133140

gacagcttac gtcattattt cttctatgta tttctcggag agatcaaaga ctttaatact 133200

ttcacttatt ctgctaccgt tatctagaag gcggagccag gtgtgcagag cggaacgtga 133260

aagtggacca ggagcgtgac cgctgaagca cagcatcaca gggagacgtt taggccacca 133320

gacggctgcg ggcgggcttg actgatgtca ggctttccac aagaggtggt ggagcagagt 133380

cttctctaac tcccctggag aaagggagac tccctctccg ggtttgggaa ggtaaggggg 133440

tccttcccag gcactggcgc taccgctgtg ctaagtgacg ggtgccttcc ccctggtgtt 133500

accgctggac cagggagccc tctagtggcc gtgtccgggc atgacagagg gctcacgctc 133560

ttgtcttctg gtcgcttctc accgtgtccc ttcagctcct atctctgtat ggcctggttt 133620

tccctaggtt atgattgtag aacaaagatt attataatat tggaataaag agtaatgcta 133680

caaactaatg attaataata ttcatatata atcatatcta taatctattt ctagtataac 133740

tagtcttatt ctatatattt tctttgttat actggaacgg cttgtggtga gtttctttat 133800

tataccggaa cagcttgtgc cttcggtctc ttgcctcggc acctgggtgg cttgccaccc 133860

acagaaatcc tgcccgtttc tcttggttaa atataatgta tttctagtag gcattgcttc 133920

ttctttccag agacaaaact caggaggatt gtgctatgat aaacaaggct aacctgctga 133980

ttctctgaag caaggaactg gagatggtcc ttctaagggt ttatattctg gattccagga 134040

aacatgcaaa caggacccat aaatgcatct ttattttggt gcccattttg acctggtcag 134100

gagaattcca acaaaaaatc cacggtgttg gagcaagatc tcaggctctc aggctgtggc 134160

catctagagg taagaagcac tttctgttgt cttaaagaaa agaaagtcct tttttttttt 134220

tttttttttt tttgagacaa tgtctcgctc tgtcgcccag gctggaatgc agtggcgtga 134280

tcttggctca ctgcaagctc cgcctcccgg gttcatgcca ttctcctgcc tcagcctcct 134340

gtgtagctgg gactacaggc gcccgccacc aagcccggct aattttttgt atttttagta 134400

gagacggggt ttcatcttgt taggcaggat ggtctcgatc tcctgacctc atgatccacc 134460

cgcctcggcc tcccaaagtg ctgggattac aggcgtgagc caccacgccc gcccagaaag 134520

tgctttcttt cagagggtta cggtttgaga aaagcagcat tgaagttgac gctgatcttg 134580

gtaatacatt tgcagagcat gcttatggtc agacgtggat gatggtgggg ttttgttttt 134640

gttttgtttt ttatctgaga cagggtgtct ctgttgccca ggctggagtg cagtggcaca 134700

tccaacctag atctcttggg ctcaagtggt cctctttttg aaacagagtc tcactctgtt 134760

accctggctg gagttcagtg gcaggaactt ggctcactgc aatctctgtc tcctgggttc 134820

aagcaattct ctggcctcag cctcccgagc agggttacag gcatgtgcca ccacgcccgg 134880

ctaatttttg cattttttta attgatttat ttgtttgttg ttgagacagt ctcactctgt 134940

tgcccaggct gtagtgcagt ggtgtgatct cagctcaccg caacctccac ctcccatgtc 135000

agcctcctga atacctgtct ccaggcatgc acccccacac ctggctaact tttgcatttt 135060

ttgtacagat gaggtttcac catgttgccc aggctggtct tgaacccctg agctcaagtg 135120

gtcctcccac ctcggcctcc taaactgctg tgattaatgg tgtgagccac cgtgccacac 135180

ccgtacttgt ttgtttctgc tcaaattgta ttgaggagct tttacatcct agtctgtggt 135240

ctatcctgga ggatgtttgt gtatataaca atgtacattt tcaacatttt agattccatt 135300

ttggatgctc ccatcgggac tgtgtgtccc tgtgctggaa ctcgagtgaa cacttggctc 135360

aaaatccatt gctgttctct agaaatcctg ccctattctc ttggtgaaat ataaggtatg 135420

tctagtcggc attgattttt ctttctggag acaaaactca ggagggttgc ccctgcatga 135480

acaaggctaa cctgctgagc ctttgaagca aggaactgga gatggttttt ttaggggttt 135540

atattctgga ttccagaaaa catgcaaaca gggacaatga atgcatcttt atttttctgt 135600

ccattttaac ctggtaaacg aaaatttcac caaaaaaccc agagtcctgg agcaagaaga 135660

tctcaggctg tgaccctcca aagggaagaa ctttctgttg tctaaaagaa aagaacgcac 135720

ttccctttag agtgttaccg tgtgagaaaa gcaacgttga agctgatgct gatcttggta 135780

ataagtttgc agagcctgct tatcatcaga cttggacgac ggtgtagttc tgttttggtt 135840

ttgagttttt tgtttgtttg ttttgttttg cttttgagat ggaggcttgc tctgtcgccc 135900

aggctggagt gcagtggcgt gttctcagct cactgcaacc tccgcctccc aggttcacgc 135960

cattcttctg cctcttgagt agctgggact acaggcgccc gccaccacgc ctggctaatt 136020

ttttgtattt ttagtagaca cagagtttca ctgtgttagc caggatggtc ttgatctcct 136080

gaccttgtga tccacccgcc ttggcctcca aaagtgctgg gattacaggc gtgaaccact 136140

gcgcccggcc ttaatttttg tatttttagt agagacagga ggctgagctt tcagtgagct 136200

gagttcgttc cactgcactc cagcctgggc aacagagtga gactccatct caaaaaaata 136260

aaaaataggc cgagcgtggt ggcacatgcc tgtaatccca gccctttggg aggccgagac 136320

aggtggatca cctgaggtca ggagttaaag gctggctggg ccaacaaggc aaaacctcga 136380

ctctactaaa aatacaaaat tacacctgta atcttagctg cttggggggc tgtggcagaa 136440

gaacctcttg aacccaggaa gaggagattg cagtgagacg aaatcatacc attgcactct 136500

agcctgggcg acagagcgag attctgtctc aaaaaataaa taagtagggc agacagggtg 136560

gctcacgtct gtaatcccag cactttggga ggcaaggcgg gtaaatcaca aggtcaggag 136620

ttcgagacca gcctggccaa catggcgaaa ccccatctct actaaaaata caaaaaagta 136680

gctgggcatg gtggctggca cctgtagtcc cagctacttg ggaggctgag gcaggagaat 136740

cgtgtgaacc caggaagcgg aggttgcagt gagccgagat cgggccattg cactccagcc 136800

tgggcgacag tgcgagactc tgtatcaaaa ataagtaaat aaataaacaa aataaaagta 136860

aaaagaatta aatagataaa tatggtacca ttatgaattt gagtgtcacc tttgtgcagg 136920

ggtcatggta atctttgtta tttcaatttt tttatgcgta ttgccaaagc aagcacatat 136980

gcgtaggaat tatacttcct gtgagaatac aatatatgga gctttctttt ttgttttttc 137040

tgtttgtttg ttttttgaga cacagtttca ctcttgttgc ccaggctgga gtgcaatgtc 137100

acgatctcag ctcacggcaa cctctgcctc ctgggttcaa gcgattctcc tgcctcagcc 137160

tcccaagtag ctgggattgc ttgcatgtgc caccaagcct agctaatttt ttgtatttag 137220

agtagagatg gggtttctcc atgttggtca ggctggtctc gaactcctga cctcagatga 137280

tccatccacc tcagcctccc aaagtgctgg gattacagat gtgagccacc atgcccggcc 137340

atgatatatt tttatatatg taatatcaca tacacacaaa atactcttct atgcacagag 137400

aatagtctta tatcactttg ggtatttgtg tgtgtgtgtg tgtgtgtgtg tgtgtgtgtg 137460

tgtgtgtgtg tgttttgttt ggttggttat ttttgagaca aggtctggct ctgtcatcca 137520

ggctggagtg tagtggtgtg atctcagctc actgcaacct ccacccccga cctaagccta 137580

ctgaattagt gtctacaggc acgcactacc acacctggct cacttttgta ttttttgtac 137640

acatggggtt tcacaatgtt gccggggctg gtctcaaact cctgtgctca ggtgatcctc 137700

ccatcacggc ctcctaaagt gctggaatta gaggtgtgag ccgccacgcc ccacctgtac 137760

ttgtgtatct ctgttcaaat gttactgagg agcttttaca gcgtaacctg tggtctatcc 137820

tggaaaatgt ttgtgtgtga gacaatgtac attcttcaac atcttagatt gcagtttgga 137880

tgctcccgtc aggactgtgt gtccctgtgc tgggactcaa gtgaacactg ggctctccat 137940

ccattgctgt tgtctagaaa tccagcccaa ttctcttggg taaatatgag gtatgtgtag 138000

taggcattgc tttttctttc tggagacaaa gctcaggagg attgcccctt gataaacaaa 138060

gctaacctgc tgattctttg aagcaaggaa ctggagatgg tccttttagg ggtttatatt 138120

ctggattcca gaaaacatgc aaacaggacc aataaatgcg tgcttatttt tgtgtctgtt 138180

ttaacctggt caaggaaaat tccaacaaaa aatccacgat gctggagcaa gaagatctca 138240

ggctgtgtcc ctctagaggg aagcgctttc tgttgtctga aagaaaagaa aatggttccc 138300

tttagagtgt tacgctttga gaaaagcatc gctgatcttg gtaacacatt tgcagagaat 138360

gcttataatc agacgtggat gatgttgaag ttttgcgttt gttttgtttt gttttttttc 138420

ctagacaggg tgtctgctgc ccaaactgga gtgcggtggc acttccaacc tagacctctt 138480

gggtttaagt ggccctctat tttgggatag agtcttgctc tgtggccctg gctggagtgc 138540

agtgtcagga actctgctca ctgcgacctc tgcctcctag gttcaagcga ttctcttgcc 138600

tcagcctcct gagcagctgg gattacgggc atgtgccacc atgcctggct aatttttgta 138660

ttttttatta tttatttatt tatttatttt tgagacagag tctcgctctg tcacccaggc 138720

tgaagtgcag tggtgcgatc ttggctcacc gcaacctcta cctcccacct cagcctcctg 138780

aacagctgtc tccaggcgtg caccaccaca cctggctaac ttctgtattt gttttacagt 138840

catggtttca ccatgttgcc cgggctcatc ttgaactcct gagctcaagt gatcctcccg 138900

cctcagcctc ctaaagtgct gtgagccacc gtgtcccaac agtacttgtg tgtttccgtt 138960

caaattgtat tgaggagctt ttacagcata gcctgtggtc tctccgggag aatgtttttg 139020

tttacgacaa tgtatattct tcaacatcgt agattccatt ttggatgccc tcattgggac 139080

tgtgtgtccc tgtactggaa ctcaagtgaa cacttggctc aaaatccatt gctgttctct 139140

agaaatccag cctaattctc ttggttaaag gtaaggtatg agtagtaagc attgcttttt 139200

ctctttggga acaaaactca ggaggattgc cccttgatga acaaagctaa cctgttgatt 139260

ctttgaggga aggaactgga gatggtcctt ttaggggttt atgttctgga ttctagaaaa 139320

cacacaaacc ggacaaataa atgcatcttt atttttgtgt ctattttaac ctcgtcaagg 139380

aagattccaa caaaaaatcc acagtgccgg agcaagaaga cctcaggctg tgacactcta 139440

gagggaagcg ctttctgttg tctgaaagaa aggaaagtgc atccttttag agtgttactg 139500

tttgagaaaa gcaacattga agctgatgct gatcttggta ataaatttgc agcgaatgct 139560

tataatcaga cttggatgat gttgggcttc tgagtttttt tttgttttgt tttgtttagt 139620

tttggggttt ttctgttttt tgttttttgt tttttttttt tttgaggccg agtcttgctc 139680

tttcgcccag gctggagtgc agtggcgcta tctcggctca ctgcaagttc tgcctcccgg 139740

gttcacgcca ttctcctgcc tcagcctcct gtgtagctgg gactacaggt gcccgccagc 139800

acacccggct aattctttgt atttttagta gagacggggt ttcaccgtgt tagccaggat 139860

ggtctcgatc tcctgacctc gtgatccacc cgcctcagcc tcccaaagtg ctgggattac 139920

aggcgtgagc caccgcgtcc agccttgtta ttgtttttga gactgagtct tgctctgtgg 139980

cccaggctgg agcgcagtgg cgtgatctcg gctcactgca acctccatct cccaggctca 140040

agcaattctc ctgcctcagc ctcctgagta gctgggatta caggcctgtg ccaccatgcc 140100

cggctaatta tttgtatttt tagtagagac agggtttgca ccatgttggc caggctggtc 140160

tcgaactcct gacctcaggt aatccaccca cgtcgatctc ccaaagtgct gggattacaa 140220

ctgtgagcca ctgcgcccgg cctgtttttc tctaccacag ggtgtctgtt gcccaggctg 140280

gagtgcggtg gcatttttaa catagatctc ttgggctcga gtggtccttt tttttggttc 140340

agtcttgctt tgtgacccag gctggagggc agtggcagaa actctcctca ctgcaacctc 140400

cgcctcctag gttcaagcca ttctcctgac tcagcctcct gagcagctgg gattaccggt 140460

gtgagccacc acacccagct aatttttgta tttttattta tttgtttatt tatttattta 140520

ttgttgagac ggagtcttgc tgtgtctccc aggctggagt gcagtggcgc gatcgtggct 140580

tactgcaacc tcctcctcct tggttcaacc aattctcctg cctcagcctc ctgagaagct 140640

aggattacag gcatgcacca ccatgcctgg ctaatttaga ttccattttg gatgctccca 140700

tcaggactgt gtccctgtac tggaactcag gtgaacactt ggttcaaaat ccgtcgcttt 140760

tctctagaca tccagctcag ttctcggtaa agataaggta tgtgtagtag gcattgcttt 140820

ttccctttag agacaaaact caggaggatt gccccttggt gaacaaggct aacttgcaga 140880

ttctttgaag caaggaactg gagatggtcc ttttaggggt ttatgttctg gattccagaa 140940

aacatgcaaa cagggccaat acatgcatct ttatttttgt gtccgttttg acctggtcaa 141000

ggaaaatttc aacaagaaac ccagagtgcc ggagcaagaa gatctcaagc tgtgactgca 141060

aagggaagcc ctttctgttg tctaaaagaa aagaaagtgc ttccctttgg tgaattacgg 141120

tttgagaaaa gcaacgttga agttgatgct gatctcggta atacatttgc agagcatgct 141180

tatcacactt ggacggtggt ggggttctgt tttggttttg cttttttatt ctaagacagg 141240

gtctgtgttg cccatgctgg agtgcggtgg cacatcccac ctaggtctct tggattcaaa 141300

tggtcctttt tgggggcaga gtctctctat caccgaggct ggagtgcagt gatcccatct 141360

tggctcacta caacctccgc ctcccaggtt caagcaattc tcctgcttcg gcctcccaag 141420

tagctgagat ttacaggtgc ccatccccac acctggataa tatttggttt tattcatgta 141480

tttatttatt tttaagatgg agtctcactg tcacccagac tggagggcag tggcacgatc 141540

tcggctcgct gccacctgcg cttccctggt tcaagtaatg cttctgcctt agcctcctga 141600

gtagctggaa ttacagaagt acaccaccac acctggctac tttttatatt tcatttttaa 141660

tacagactgg atttcaccat gttggctagt atgatatata ttttgtcata tattcttata 141720

tatataatca tatacacaca aaatactctt ctgcacacga agagtattct tatatcgctt 141780

tgggtttttt tgtttgtgtg tttgttttgt gtttgtttct ttgtttttga gacaacgtct 141840

ggctctgttg ctcaggctaa agtgtagtat tgcaagctca gctcactgca acctccacct 141900

cccacctaaa gtcccctgaa ttagtgtcta caggcatgca ccaccacacc tggctcactt 141960

ttttttctga gaaggaattt tgcttttgtc gctcaggctg gagtgcaatg gggtgatttc 142020

agctcactgc aacctctgac tcccggcttc aagcaattct cctgcctcag cctcccaagt 142080

agctgggatt acaggcatgc accaccacac ctggctaatt ttgtattttt agtagagatg 142140

ggtttcacct tctttctcag gctggtcttg aactcctgac ctcaagtgat ccacctgcct 142200

cgacctccca aagtgctagg attacaggtg agagacattg cacctggcca cctaactttt 142260

gtagtttttt tacagacagg gcttcaccat attgcccaag ctggtctcca gctactgagc 142320

tcaagtgatg ctcctgtctc ggccactgtc cctcacccaa acttgtgtat ttcttttctt 142380

ttcttttttt tttttttttt ttgagatgga gtctggctct gtcacccagg ctggagtgca 142440

gtggcacgat cttggctcac tgcaagctcc gcctccccag ttcatgatat ttttctgcct 142500

cagcctccca agtagctggc actacaggca cccgccacca cgcctggcta attttttgta 142560

tttttagtga agacggggtt tcactggatt agccaggatg gtcttgatct cctgagctcg 142620

tgatctgcct gccttggcct cccaaagtgc tgggattaca ggcttgagcc accgcaccca 142680

gcttttttgt ttttttgttt tgtttttttg agatggggtc tcgctctgtc acccagcctg 142740

gagtgcaatg gcacgatctc ggttcactgc atcctacccc tcccgggttc aagcgattct 142800

cctgcctcat cctcccaagt aactgggatt acaggtgccc accaccgcgc ctggctaatt 142860

ttttgtattt ttagtagaga cggggtttca ccgtattagc caggatggtc ttgatctcct 142920

gacctcgtga tctccctgcc tcagcctccc aaagtgctgg gattacaggt gtgagccact 142980

gcacccggcc tttttttttt tttttttttt tttgagatgg gatctcgctc tgtcacccag 143040

gctggagtgc aacggcatga tctcagctca cggcaacctc cccctcccgg gttcaggtga 143100

ttctactgcc tcagcctccc aagtagctgg gattacaggt gcccacgacc acgcctggct 143160

aattttttgt atttttagta gagatggggt ttcatcaagg tggccaggct ggtctcaaac 143220

tcctgacctc atgatctact ggcctcggcc taccgaagtg ctgggattac aggcatgagc 143280

caccgcgccc agcttctgta tttcttttca aattttattg aggatttttt atgtcctagc 143340

ctgtggtcta tcctggagga tgtttgtgtg tgagacaatg tatattcatc aacatcttag 143400

attccatttt gattgcttcc gtagaaactg tgtgtccttg tgctggaact ccggtgaaca 143460

cctggctcaa tatccattgc tcttctctag aaatccagcc cagttctctt ggttaaatat 143520

aagatatgtg tagcaggcat tgctttttct ttccagagac aaaactcagg aggattgccc 143580

cttgatgaac aaagctaacc tgctgattct ttgaagcaag taactggaga tcctcctttt 143640

aggggtttat attctggatt ccagaaaaca tgcaaacagg gccaataaat gcatctttat 143700

gtttctgtcc attttaactt gatctaggaa aattccaaca aaaaacccac ggtgctggag 143760

caagatctca ggctgtgacc ttctcgagga aagaagcact ttctgttgtc tgaaagaaaa 143820

gaaagtgctt cctttcagag ggttacggtt tgagaaaagc aacgttgaag ttgacgctga 143880

tcttggtaat acatttgcag agcgtgctga tcatcagacg tggataatgg tggggttttg 143940

ttttttatct aagacagggt gtctgttccc caggctggag tgcggtggca cttccaactt 144000

agatctcttg ggttcaagtg gccctctttt tgggacagag tctcactctg tggccctggc 144060

tggagtgcag tggcaggaac ttggctcact gcaatctctg cctcctgggt tcaagcaatt 144120

ctcctgcctc agcctccgga gcagctgggg ttacaggctg tgccaccatg cccagctaac 144180

ttttgcattt ttattattga tttattgatt tatttttgag acagggtctg gctctgtcac 144240

ccaggctata gggcagtggt gcgatctcgg ctcactgcaa cctccacctc ccacctcagc 144300

ctcctgaata cctgtctcca ggcatgcact accacacctg cctaactttt gtattttttg 144360

tatagatgag gtttcaccat gttgcccagg ctggtctcgg acccttgagc tcaattgctc 144420

ctcccgcctg ggcctccaaa actgctgtga ttaatggtgt gagcaagcgt gccctactct 144480

tgtgtgtttc tgctcaaatt ttatggagga gcttttacat cccagcctgt ggtctatcct 144540

ggagaatgtt tgtgtgtgtg acaatgtata ttcttcaaca ttttagattc catttggatg 144600

ctcccatccg gactgtgtgc ccctgtactg gaactcaagt gaacacttgg ctcaacatcc 144660

attgctgttc tcatgaattc caggccaatt gtcttggtta aagacaaggt atgtgggctg 144720

ggcgtggtgg ctcatgcctg taatcccagc actttgggag accgaggtgg gaggatcacg 144780

aggtcaggag ttcgagacaa gcctgaccaa cacggtgaaa cccatctcta ctaaaaatac 144840

aaaaattagc tgggcgtggt ggctccacca gtaatctcag ctacttggga ggctgaggca 144900

ggagaattgc ttgaacccag gaagcggagg ttgcagtgag ctgagatcgc gccactgcac 144960

tccagcctgg gtgacagagc gagacttcat ctcaaaaaat aaaaaataaa aaggtatgtg 145020

tagtaggcat tgctttttct ctttgtagat aatactcagg aggactgcct cttgaacaag 145080

gctaacctgc tgagcctttg aagcaaggaa ctggagatgg tccttttagg gggttatgtt 145140

ctggattcca gaaaacatgc aaatagggac aatgaatgca tctttatttt tctgtccatt 145200

ttaacctggt caaggaaaat ttcaacaaga aacccagagt gctggagcaa gaagatctca 145260

agctgtgagt ctacaaagga aagcgctttc tgttgtctga aagaaaagaa atcgcttccc 145320

tttggagtgt tacggtttga gaaaagcagc gttgaagttg atgctgatct cggtaataca 145380

tttgcagagc atgcttatca cacttggacg gtggcggggt tctgttttgg ttttgctttg 145440

ttattctaaa acagggtctg tgttgcccat gctggagtgc ggtggcacat cccacctagg 145500

tctcttggat ttaaatggtc ctttttgggg gccagagtct ctctctgtca ccgaggctga 145560

agtgcagtca tcccatcttg gctcactaca acctccgcct cccaggttca agcaattctc 145620

ctgcttcagc ttcccaagta gctgagattt acaggcgccc atccccacac ctggataata 145680

tttgggtttt ttttttattt atttttaaga tggggtctca ccgtcaccca ggctgtagtt 145740

cagtggcttg atctcggctt actgcaatgt atgccaccca ggatcaagtg attctcctgc 145800

cccagactcc tgagtagctg gaattacacg cacctgccac tacacccggc tactttttga 145860

attttttttt ttttttttaa agtaaagatg ggctttcacc atgttggcca ggctggtctc 145920

aaacccctga cctcaagtca gctgcttgcc tcagcctccc aaagtgctga gaatacaaac 145980

atgagccact gcgtggccat caacacctct tactttatgg aaatttttct ggcactggta 146040

tagaacctca cgtggggtca ggtggagttg aggggacctc agtttccctt gcagatggga 146100

tgtgcactgc tcagcaagag cacagaggtg gagtgcatgg ggcttcagtg tttattgggg 146160

aaatgaagct aaaatcttcg gtgtgtgacc aggagataaa tgcatgagat ggggatctca 146220

ctatgctgcc caggctgaag tgggcttaga tcctcctgcc tctgcccctc cccagtcctt 146280

ggtacatggg actacatgtg aatattaacc ccccatgcac agacaagaag aaagtaagga 146340

ctgttctttg gttcatacct gaccccagtt aaacatgtat tttagataaa caatgtattt 146400

gaaatgtact tgaacaacaa atgatttgct gtttaggtgt gggcatcttt ttttttcttc 146460

ctaactttaa atatgggact agtccaggta cggtggctca tgcctgtaat tccagcactt 146520

tgggaggcgg agacaggagg atcacctgag gtcagttgtt cgagaccaga ctggccaaca 146580

tggtgaaacc tcgtctgtac taaaaataaa aaaattagtg agacgtggtc ttggtgcatg 146640

cctgtaatcc cagctactcg ggtggatgag gcaggagaat tgcttgaggc tgggaggctg 146700

agctttcagt gagctgagat caagcctctg cactccagcc tgggcaacag agtgagactg 146760

agtctcaaaa aagtaaaaaa tcagctgggt gcgctggcaa acacttgtaa tcccagcact 146820

ttgggaggcc gagacgggag gatcacctga ggtcatgttc aagactaaca gggctgaaat 146880

agcaaaacct catctctatt aaaaatacaa aaattagctg ggtgcagtgg tgtatatctg 146940

tatttccagc tacttggagg ctaaggcagg agaatcgctt gaacccagga ggaagagatt 147000

tcagtgagct gaagccatgc cattgcactc cagcctgggc cacagagcaa gattccatct 147060

caaaaataaa ataaataata aaaatgaaat taaatagata aaaataaata gataaatatg 147120

agaccatcat gaatttggat gtcacttttg tgcaggggcc agggtaatct ctgtcattcc 147180

aatttttttt atgtgcactg ccaaagcaag cactcatgtg taggaattat tcttcctgtg 147240

agcatataat atatggaact ttgagacaga gtctcaaaca tatataatgt atattttatc 147300

atatattctt tatatataaa atatcatata gacacaaaat actgttctaa atacagagaa 147360

tactctgata tcaacttggg tattttttgt ttctgtgtgt gtgtattttg tttggttggt 147420

tatttttgag acaaggtcta gctctatcac acaggctgga gtgtagtggt gcgttctcgg 147480

ctcactgcaa cctccgcctc ccacctaagc ctgctgcatt agtgtctaca ggcatgcacc 147540

accacacccg gctcactttt gtattttttg tacacatggg gttacaccat gttgccgggg 147600

ctggtctcga actcctgagc tcaggtgatt ctcctgtctt ggcctcctaa agtgctggga 147660

ttagaggtgt gagccaccat gccccatcgg tacttgtgat ttaggttcaa attttgatga 147720

gtttttatga ccaagcctgt ggtctatcct agagaatgtt tgtgtgtttg acaattcaac 147780

atcgtagagt tgattttggg tgctcccatt gggactgtgt gtccctgtac tggaacttga 147840

gtgaacactt ggctcataat ccattgctct tctctagaaa tccagcccaa ttctcttggt 147900

taaatataag gtatgtgtag caggcattgc tttttccttc cagagacaaa actcaggagg 147960

attgcccctt gatgaacaag gctaacctgc tgattctttg aagcaaagaa ctggagatgg 148020

tccttttaga ggtttatatt ctggattcca gaaaacatgc aaacagggcc aataaatgca 148080

tctttatgtt ttcgtccatt ttaacttgat ctaggaaaat tccaaaaaaa aaaacccacg 148140

gtgctggagc aagaagatct caggttgtga ccttctcgag gaaagaagca ctttctgttg 148200

tctgaaagaa aagaaagtgc ttcctttcag agggttacgg tttgagaaaa gcaacgtcga 148260

agttgacgct gatcttggta atacatttgc agagcgtgct gatcatcaga catggataat 148320

ggtggggttt tgtttttgtt ttattttttt atctaagaca aggtatctgt tgcccaggct 148380

ggagtgcggt ggcacttcta acctagatct cttgggttca agtggccctc ttttcgggat 148440

agagtcttct ttgctctgtg gccctggctg gagtgcagtg gcaggaactc ggctcaccgc 148500

aacctctgcc tcctgggttc aagcaattct cctgcctcag cctgtcgagc agctggggtt 148560

acagacatgt gccaccacgc ctggctaatt tttgcatttt tattgtcgat cgattgattg 148620

atttttgaga cagagtctgg ctctgtcacc caggctgtcg ggcagtggtg cgatcttggc 148680

tcactgcaac ctccacctcc cacctcagcc tcctgaatac ctgtctacag gcatgcacaa 148740

ccacaccttg ctaacttttg tattttttgt acagatgagg tttcaccatg ttgcccaggc 148800

tggtctcgga cccctgagct caagtactcc tcaaacctgg gcctcctaaa ctgctgtgat 148860

taatggtgtg agccaccgtg ccctactctt acttgtgtgt ttctgctcaa attttattaa 148920

ggagctttta tggcctcgcc tgtggtctgt cctggaggat gtttgtgtgt gtgacaatgt 148980

atattcttca acatcttcga ttccattttg ggtgcctcca tggggactgt gtgtccctgt 149040

actggaacgc aagtgaagac ttggctcaga gtccatttgc tgttctctag aaatccagcc 149100

taatcctctt gtgcaaatat aatatatatc tagtaggcat tgctttttct ttctggagac 149160

aaaacacagg aggattgccc cttgatgaac aggactaacc tgctgattct ttgaagcaag 149220

gaactggaaa tggtcctttt agggatttat gctctggatt ccagaaaaca cgcaaacagg 149280

gccaataaat gcatctttat ttttgtgtcc attttgacct ggtcaaggaa gattccaaca 149340

aaaaatccac agtgccggag caagaagatc tcaggctgtg tccctctaca gggaagcgct 149400

ttctgttgtc tgaaagaaag gaaagtgcat ccttttagag tgttactgtt tgagaaaagc 149460

aacgttgaag ttgatgctga ttttggtaat acatttgcag agcatgctta tcatcagact 149520

tggatgatgt tgggttctgt ttttgctttg tttttttttc caagacagtg tgtttgttgc 149580

ccaggctgga gtgcggtggt acttcccacc tagatctctt gggctcaaga ggtctttttt 149640

tatttttctt tctcaagaga gagtctggtg gtgacaccca ggctggagtg cagtggtgca 149700

ttatcagctc actgcagcct tcccctcccc ggttcaagtg attctttcac ctcagcctcc 149760

cgagtagctg ggattacagg tgtgggctac cacacccggc taatttttgt atttttagca 149820

gagacagggt tttaccatgt tggggaggct ggtctcaact cctgtcctca agcgatccac 149880

ctcccttgcc tcccaagtac tgagattaca ggcgtgagca actgcgcccg gcctcaagtg 149940

gtcctcttaa gtcagcctac caagttttgg gactacatgg ggcatgccac cacacttggc 150000

taagttttta attttttttt tttttttttt tttttttttt gagacggagt ctcactctgt 150060

cgcccaggct ggagtgcagt ggcaagatct cggctcactg caagctcggc ctccggggtt 150120

cacgccattc tcctgcctca gcctcccgag tagctgggac tacaggtgcc cgccaccacg 150180

cctggctagt tttttgtatt tttagtagag acggggtttc accctgttag ctaggatggt 150240

ctcaatctcc tgacctcgtg atccacccgc ctcggcctcc caaagtgctg ggattacagg 150300

cgtgagccac cgcgcccggc cgatagtttt taatttttga tagaaaggga atctctcttg 150360

cctaagatgg tctcaactcc tgagctcaag ggatcctaaa ggtgtgagcc gccttgtcct 150420

gatgacccat ttcaaacgta gctgacatgg ccaggcatca tggggcacac agtcccagct 150480

actgcagaag ccggggtggg agggtccttt gatttccagg ctataccatg tgctgatcac 150540

acttttgatc ccgagtagct gggattacag gcagccaccg ccaggccggc taatttttat 150600

ttatttactt attttttcag acggtgtttc cctcttgttg cccaggctgg agtgcaatgg 150660

catgatctcg gatcactgca acctccacct ccctggttca agcgattctc ctgcctcagc 150720

ctcccgagca gctgggatta caggcatgca tcaccacgcc cggctaattc tttgtatttt 150780

tagtagagat gggatttctc catgttggtc aggctgatct tgaactccca accttaggtg 150840

atccacccac ctcggcctcc caaagtgctg ggattacagg catgagccat tgtgcccggc 150900

ccatttcatt tatttttatg tgtgctgctg aagcaagcac ttatgtgtag gaattgttct 150960

tcctgtgagc atatgttggc cagcctggac cacataccaa gatcccatct ctttaaaaac 151020

acagattacg tggcacctgg cacctggtcc cagagacttc atttgggttg gtcatttgaa 151080

acactagcct cccatcaatt tagtgtaatc aatccaaatc atgtgtcctt cattaagaga 151140

ctaagaacgc ctccacgtct atccagtcta ttttgtaatc cccaacggtt gtcaatatta 151200

ataaaatttc ttttcttttt cctattcatt tgtgtcttta gtttttcttc cccaaaaact 151260

tgccatcatt tctcggaata gacctgcttt ctctgcagga agggtgtggt tgattcaacc 151320

cttacccact aatgccaacc ccagtgagtt cttttatcct attttctatg ggtaacgatt 151380

ccaaggtacc attccaccgg gcaaaagctc acatctaagt gtcagttgct cggtaagatg 151440

tgaaatgttt gctgtggcta atataagaaa caaactattg tagacaagat gaatctcagt 151500

ggcagtgata aatgtcgcac aagacaaaac cacagatcct tttttttttt tttttttttt 151560

ttagacaaag tttcactctt gttgcccagg ctggagtgca atggtgcgat ctcggctcac 151620

tgcaacctcc gcctcctgga ggcgattcaa gcgattctcc tgcctcagcc tcccaagtag 151680

ctgggattac aggcatgcac caccacgccc aggtaatttt gtatttttag tagagacggg 151740

gtttcgccat gctggtcagg ctggaactcc tgacctcaga tgatcctccc acctcagcct 151800

cccaaagtgc tgggattaca ggcgtgagcc actgtgcccg gccaaccaca gatactttca 151860

cgaaagcctt tagggcccta gaaggggctc cctagtaaca ggtgggatgc gaggcagctc 151920

tcgttgttgc cgtagtgagc gatgcctgtt cgtccagccc tcaacacctt ttactccgtg 151980

gaagttatgc ctgcactggt ttacagaacc tcccttgact tgggttgagg tagagctgaa 152040

gggaacctca gtgtcccttg caggtgggat gtgcactgct tagcaagagc acggaggtgg 152100

agtgcatggg ctttgagttt ttattgggaa aatgaagctg aaatgtaggg cgcatgacca 152160

catcataaat gcacatttga tttaattttt ctattttatt tttatttatt tattttgaga 152220

tggagcctcg ctctgttgcc caggctggag tgcactggcc tgatctcggc tcactgcaac 152280

ctccacctcc cgggttcaag caattattgt gcctcagcct cccaggtagc tgggattaca 152340

cgcatgcttc cacgcccggc cgatttttgt atttttagta gagacggggt ttcaccatgt 152400

tggccaggct ggtttcaaaa tcctgacctc aagtgatccg cccgcctcgg cctctcaaag 152460

tcctgagatt acaggcgtga gccaccatga ccagcctaat ttttctattt tagagacagt 152520

ggtctagcta tgccacccgg gctaaactag gctttagaga tcttcctgcc tctgcccctc 152580

cacggtctct ggtagtttgg actacaaatg aacattagca tatttgcaac caccaccact 152640

ctagtttgga agatttttat caccccaaag aagcttatac ccatttgcct tcagtaccca 152700

cccccctctt ccactcagac cctggcaact actctacatc tctctagctc tggatttgcc 152760

tcttgtgggc atttcacaaa aaccagtctt gaatgggtgg cctgttgtga ccacttttaa 152820

tataatgggt tggttctggt tgtctgaagt gtgaatcttg ccaatgaagg atggtccctg 152880

gatggaagca ggaggctggg agaactgggc ggaacatcct ttcggaatgg agtggggtgg 152940

gcacaccctg atgtctggga agctcacaag ggtggaagaa cccatcttcc tctctgataa 153000

ctgcaaggtg accctcctgg ggcactggat ggagtgaagg catctggact gggaacacca 153060

gggcattgca ctggtgcagg caggatgagc cgaggggaaa ggagtgccag gcatcattct 153120

ctggtgacag tttgggtttg atctggatgg agcaggtgtc ttctggtaga gagagtccct 153180

gggatttttg ctctgctcct ggctgtcttt cagtcatgga atctgatgac aaaggctccc 153240

actctgggcc acttcatttg gtttctggag cccagtggtc ctttctgccc ggactcagga 153300

tcttttgggg aaatttggga cctcgcagga catctgcaca atccatagaa atccctgaga 153360

gccccttccc tttgctgaca tctccgtatt cctacctatt gccttccaaa aaaagaccct 153420

tatctgaatt gccaaagggg gcttcccaga gcagggaaac ccggttaaat ttgtatttca 153480

gattaacagc atatctagaa tctactcgaa caagaagtga tttgttgttt aagtgtgaga 153540

aatttttttc ccctaatttt aaatacggaa cccatcatga atttggctgt cacctttgcg 153600

caggggccgt gggaacctat catttcattg tttaatgtgt gctgccaaag caagcactta 153660

agtgtgggaa ttattcttca cgtgaggata caatagatgg aacgttatta cttttttctt 153720

tcataattga gattttattg gttgaagatc ggtacagaca tttcaatttg tacacaattc 153780

ttaacatacg taccgaaaat ctaaaaagcc atgtattgta aatcgttttg ttttgttttg 153840

tttttttgag acagagtttc gctcttgtca cccaggctgg agcgcaatga cgcaatctcg 153900

gctcaccgca acctccgcct cccgggttca agcaattctc ctgcctcagc ctcccaagta 153960

gctgggatta caggcaaatg ccaccatgcc cggctaattt ttgtattttt ttttttttaa 154020

gtagagacag ggcttctgcc tgtcggtcag gctggtctcg aattcccgac ctcaggtgat 154080

ctgcccgcct cggcctctta aagtgctgag attacaggcg tgagccaccg cacccggcca 154140

tgtattgtaa ttcttctaca aagttattcc ggcgactttc cagcttaaaa tttggaagca 154200

cattttcctt aagaggctat caagtaccag tatcttcaca tgttgatcag ctgttacgga 154260

cgtccctcca attcacaact aaaaatagca tgtaccctac atattcaaat ttttcatctt 154320

tcacaacgca gaaacaaact tattaggaga acagaactac cacaatcaaa gatgttacag 154380

agtccacaca attctaacag ggagagccat ggtcagggag tggttttctt taggaaacaa 154440

ttccaaaata cgacaggaga atagaagtaa tttaaaatgt tcaagacact aaatgcagag 154500

ctgattccat gctgccattt aatatgcttt gtattatagg atataaacac gaaccctggc 154560

cgggtgcagt ggctcacgcc tgtaatccca gcactctggg aggccgaagt gggcagatca 154620

cctgaggtca ggagttcaag accagcctgg cccacatggt gaaaccccat ccctactaaa 154680

aatatgaaaa ttagccaggc atggtggtgc gcgcctgtaa tcccagctac tccggatgct 154740

gaggcatgag aatcgcttga acccgggagg tggagattgc agtgaactga aatcacgcca 154800

ctgccctcca gcctgggcga caagaacaag actctgtttc aaaaaaaaaa aaaaaaaaaa 154860

agtttctgga actactaaaa aacttgcatt tacaaaatag ttgataaaaa gattcctctg 154920

ggttttacaa gaagcgagac agggagcact gatgagacgt ggtatacggt gaatcagact 154980

gggcatcaga ggctgggcct cctcagtttt cctttcccca ttttctgcag ataaatcttt 155040

agtttcttgg ttagccactt ctgcccgttt tccctttgct cccctttgcc cttctgttca 155100

cacttttttg tctgaagatg tatccttcgt tgctgccttt ttcggcttca tttccactgg 155160

tttttttgtt ttgttttgtt ttgttttgtt tgagacgtag tttcaccctg ttgctcaggc 155220

tggagtgcaa tggcgtgatc tcggctcaca gcaacttctg ccttccggtt caagcgattc 155280

tcttgcctca gcctcccgag tagctgggct tataggcgcg caccgccatg cccggcttat 155340

ttttgtattt ttagtagaga cgggggtttc accatgttgg ccaggctggt ctcgaactcc 155400

tgacctcagg tgatccgccc gcctcagcct cccaaagtgt tgtgattaca agcgtgagcc 155460

accgcacccg gccttgcttc cacttttgca ggagcaggtt tagctgacaa ccgtgcgatc 155520

tcctcgtggg ctcttccttg gcggccccta tggcggagct aacctgcctc ttgggcatcc 155580

tggtggcgga gagggcgcgt gccggctgtc tgcgggccgc ggctgccgag agccttggcg 155640

aagctgggct gcctggcggc tgcggctcct cccgccgccc gagctgctga gacccacagc 155700

ggggtcggtg ggagaaccga atggaacccg aaactttctt attttgagac aggatctggc 155760

tctgtcaccc aggctggagt gcttggctca ctgcaacgtc tgcctcaagt gatccttcca 155820

cctcaacctc ccaagtagct gagattacag gcgcgcgtgc cactatgcct ggctaatttt 155880

tgtactcttt gtagagatgg gggtctcgct atgttgccca cgctggtctt gaactcctgg 155940

gctcaaatga tcctcctggc ctggcttccc aaagtgctgg ttgtaacgcc agcagtttgg 156000

gaggccacag tcggcgggat ggattagcta gcaacaaaca ttccaggcag agaatggtgc 156060

tctctcaact ctcagtcagc aaggggctag tatatattga aggggctcag ggcatgtcac 156120

cccaaagtca tcgcgtcagc atatggatta tttcgacctg gaagcctgtg ggaaaaagca 156180

gatgcatctg acctccccct tcctacgtaa aagtagattg taaaatttat caagaggaaa 156240

acgcccttcc tgcgccagga agagaagaat gttcctatca gcaggggcca ggagtccata 156300

tgctaaacaa ctcagctact aacccttatc ttccttaagt tccccactgt ttcctggtca 156360

cttcccctag cccaagcctc tttgtcttgt cacatcccca caacttatca ttcctttttt 156420

ttttttgagg tggagtttag ctcttgttgc ccaggctaga gtgcgatggc gcaatcttgg 156480

ctcactgcaa cctctgcctc ccgggttcaa gtgattctcc tgcctcagcc tcccaagtag 156540

ctgggattac aggcgcccac caccacgctc ggctgatttt ttgtattttt agtagagatg 156600

gggttttgcc atgctggcca ggctggtctt gaactcctga tctccggtga tccgcccgcc 156660

tcagcctccc aaagtgctgg gattgattac aggcgtgagc caccgcgccc ggccatcatt 156720

ctcttttttg ttttgttgtg gttgttgttg tttgagacag cgtctggctt tgtcacccaa 156780

gctggagtgc agtggcgcca tctcagctca ctgcgacctc tgcctcccgg gttcaagcaa 156840

ttctcatacc tcagcctccc gagtagctgg aaccacaggc atgagccacc atgcccggct 156900

agttacaggg ttttcctatg ctctccaggc tggagtgcag tggcgcaatc atagttcact 156960

gcagtcttca actcctgggc tcaagcaatc ctcccacctc agcctcttaa actgctggga 157020

ttacaggtgg tagccatcat gccctgcccc agagaagtct ttaggcttca gccaggcacg 157080

gtggctcagg ccggtaaacc cagcactttg ggaggctgag gtgggaggat cgcttgtgcc 157140

caggagttga aggctgcaat cagctatgac tgcaccacag cactccagtc tgggcaacaa 157200

agtgagaccc tgtctcaaaa aacaacaccg taggcccagc acggtggctc ctgcctgtaa 157260

tcccagcact ttgggaggcc aaggcaggtg gatcacttga ggtccagagt tcaggaccag 157320

catgaccaac atggtgaaac cccgtctcta ctaaaaatat acaaattagc caggtgtggt 157380

ggcagacacc tgtaatccca gctacttggg agactgaggg aggataattg cttgaacccg 157440

ggaggcggag gttgcggtga gccaagatcg cgccattgca ctccagcctg gttgacagag 157500

caagaccctg tctcaaaaag aaaggaaaag aaaaagaata caccagaagg tacccagaga 157560

ggccagtgtg gatgtacagc cagcacataa gatgctggtt agggacaggc gcagtggctc 157620

acgcctgtaa tcccagcact ctgggaggcc caggcagggg gatcacctga ggtcaggagt 157680

tcaagaccag cctggccaac atggcgaaac cccgtctgta ctagcaaata caaaaattac 157740

ttgggcgcgg tggcacgtgc tataatccca gctactccgg aggctgaggg tggaaaattg 157800

cttgaacccg ggaggctgag gttgcagtga gccgagatcg caccactgta ctccagcctg 157860

ggcaacaaga gcgaaactcc gtctcaaaaa attaaataaa taggccgggc gcggtggctc 157920

acgcctgtaa tcccagcact ctgggaggcc gaggagggcg gatcacgggg tcaggagatc 157980

aagaccatcc tggccaacat ggtgaaaccc cgcctctact aaaaatacaa aaacttcgcc 158040

ctgtatggcg gcacacggct gtattcccag ctactcggga ggctgaggca ggagaacggc 158100

ttgaacccgg gaggcagagg ctacagtgag ccgggatcac accgctgcac tccggcctgg 158160

gcagcaagag caaaactctg tctcaaaaaa caaaaaacag aaaaacaaaa agaatcaagt 158220

aagtcgaagc cacactgata acagccaatt tttgtgaacc aagggagtgt caattcaaga 158280

atttacatag atgtctactt ttgctatctc ctatgtgcca agcaagatac aggctctggg 158340

caatcagaaa caaaagagac tcactcgttc ctctcacagt actcagtcct tactgagata 158400

aggacaaaag aaaatgtcct gtctggaatg cagggaaacc agaacttcag gtcaggggac 158460

atttccattg aattgtgtgg agttgaagct gaaaatactt tttttttttt tttttttttt 158520

acatcacggc atggtttatt acgtgatttt tttactatac aaacaaaaaa tacagaaatg 158580

caatatgtga atacagctaa atgcagaatg gtgacttttt tctcttcaag aggccatgat 158640

tcccatttct agtaaaataa agagaccgca tacaggttgg ttgtgagatt cacaattttg 158700

cctagaaatg atctataaat gcatttttcc cccctgctac ctaccgtaaa tcgtaaaaag 158760

ggagttaaag caaagtttcc ttgttggttc ctaccatatg gaagatgcta tattctattt 158820

tagcagggtc aatatttgga aaatatctaa attaaatatt attacaaaaa tgaagctgta 158880

atgagattct ggctaaagag ggcactaaat gagaataata tatatttaaa gaatccaaaa 158940

caaacaaaca aaaagaggtt attataaaaa gctctaggtg cactgtaagc atatagggtt 159000

tttttttcat gtgttttttt ttaaacaatg gaagtgtcaa aaatagggtc aactgtgtta 159060

gactaaatta cattattgta tatgctgcat tgaatggaac ctttgtatta taattatcat 159120

agagaagcac agtttgcatc atattatggc aattcatcgt caatgaaaac cttccagagt 159180

ccttttattt tggaatctcc tgtaaactat caaaccacca gaacatgact gtacaacagt 159240

aaaatgttct cttgcattaa actgaagaga cctgtttaat aaaaaaagaa aaagaaaatg 159300

taggaaaggt acttagagct gttactttct aagtacacaa caccctagac aattcgaggc 159360

atcttaatct ccatcaagaa caacaacaac aaaaataatt tttgtcatgt tgttaaatcc 159420

atcattatgg atcaaactgg tgcaacttgg tcaaatgaat ccaacaaaca ctgatgtcca 159480

agctggcata ttggcaacta atacacaact ggtggtcaat agagagttta aaagatcttc 159540

cctttcttgt tgttcttttc caaggttctc aaagagttct gttgctctaa aatacgttgt 159600

tgagcttccc agtttgcttt ctcatcctca aactgacgac gtttctcctc taattctttg 159660

tgccgtgctc ccaaattctt tttcagttgc acatggcact gctggagcta aatcacagca 159720

caatatatac cccaaggata ctgccttcct ctgacccttt ttccattaac ttcagtgata 159780

atatcactac ccaccacagc aagaggtaaa cggtccttta tctttttaac acgtttcttt 159840

tcttcttcat catctgtttc tggaaattca tatattttaa ttttatgttc ttggatttct 159900

ttcattatct gttttgtaaa ctgttgacat tcctctggtg tgagtgtgtc tgctttggca 159960

gtaagtggga tgatattcac tttttcatgc aaatgtttca taaactcaac atccagtggt 160020

ttaagtctca acaaaaggca atgtcaagta gttacaaaca tgcactaccc aatcctgcct 160080

gaaggtggga gtcttcagtt atgtaatggt cctgttatca ggcatctgac gtctgctcac 160140

tcgcgattct gcatttaggt agtcctcaaa tttactatca atgtaatcga taacaggctg 160200

ccagcaatta ctactatcca ctgcatctcc aaatcctgag gtatcaacta tcctgagcag 160260

caactaaaca ccaccttctt tgattaaaac tttggattgt tccatccagg cttctatttt 160320

cccatagagg tccaaaaaac gcccttgttt gttaagttga tatatgtaaa ccttttctcc 160380

aggctgttcc ttgaggttta cataattggg aactccttca agcatctgta aataaacctt 160440

ttcagctgta gatctctcta ttttcagtta atttgcagac caccaactat aggagccaag 160500

ttgaaagaga aaaaagtttc tcttcccagc actttcttta ggatgctttt cattgcttcc 160560

caactggagg cctcgcccct ccacccacct gccctagtcc tcagctgcct ccagttctgt 160620

ccacttaaat acagactccc ggtagtagag aactttttta aaattttatt attattatta 160680

ttattattat tgacacagtc ttactctgtc gtccaggctg gagagcagtg gcacaatctc 160740

agctcactgc agcctctgcc tcctgggttc cagcgattct cctgcctcag cctcctgagt 160800

agctgggatt acagacaccc gccaccatgc ccggctaatt tttgtatttt tggtagagac 160860

gggggtttca ccatgttggc caggctggtc ttgaactcct aacctcaggt gatcctccta 160920

cctcggcctc ccaaagtgct gggattacag gcgacagcca atgtgtgcac 160970

<210> SEQ ID NO: 61

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 61

actgggacga catggagaaa a 21

<210> SEQ ID NO: 62

<211> LENGTH: 14

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 62

gccacacgca gctc 14

<210> SEQ ID NO: 63

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 63

tgcaaggaaa gcattgaaca a 21

<210> SEQ ID NO: 64

<211> LENGTH: 23

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 64

gaggagtcac ctggacaatc act 23

<210> SEQ ID NO: 65

<211> LENGTH: 22

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 65

cacatgaagc agcacgactt ct 22

<210> SEQ ID NO: 66

<211> LENGTH: 22

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 66

aactccagca ggaccatgtg at 22

<210> SEQ ID NO: 67

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 67

gcggtggttg cccaacagga 20

<210> SEQ ID NO: 68

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 68

acgacccgtg gtcatcttta 20

<210> SEQ ID NO: 69

<211> LENGTH: 23

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 69

tagaggatca cgtaattgca gga 23

<210> SEQ ID NO: 70

<211> LENGTH: 23

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 70

gttgtcaaag ctgagccttc tat 23

<210> SEQ ID NO: 71

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 71

atgccagacc gtcttgatac a 21

<210> SEQ ID NO: 72

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 72

tgacccaagt atttcagccc a 21

<210> SEQ ID NO: 73

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 73

gaacaacggt ttcgctcttt g 21

<210> SEQ ID NO: 74

<211> LENGTH: 23

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 74

gcttctacat taggccagac ttt 23

<210> SEQ ID NO: 75

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 75

acccgcttaa cagcgtcaac a 21

<210> SEQ ID NO: 76

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 76

ccaaagaggt gcgggagttt 20

<210> SEQ ID NO: 77

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 77

gccaatgagg gttcgagttc 20

<210> SEQ ID NO: 78

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 78

aacaacatcc cgtcgttcat c 21

<210> SEQ ID NO: 79

<211> LENGTH: 21

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 79

acgaatccca gtgtgttttg g 21

<210> SEQ ID NO: 80

<211> LENGTH: 22

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 80

tgctcaaaaa cggtatggac at 22

<210> SEQ ID NO: 81

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 81

caaccaagca aatgtgagga 20

<210> SEQ ID NO: 82

<211> LENGTH: 20

<212> TYPE: DNA

<213> ORGANISM: Artificial Sequence

<220> FEATURE:

<223> OTHER INFORMATION: Synthetic oligonucleotide

<400> SEQENCE: 82

ggagacttgc ctggtgaaaa 20

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Patent Valuation

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23.42/100 Score

Market Attractiveness

It shows from an IP point of view how many competitors are active and innovations are made in the different technical fields of the company. On a company level, the market attractiveness is often also an indicator of how diversified a company is. Here we look into the commercial relevance of the market.

60.0/100 Score

Market Coverage

It shows the sizes of the market that is covered with the IP and in how many countries the IP guarantees protection. It reflects a market size that is potentially addressable with the invented technology/formulation with a legal protection which also includes a freedom to operate. Here we look into the size of the impacted market.

73.3/100 Score

Technology Quality

It shows the degree of innovation that can be derived from a company’s IP. Here we look into ease of detection, ability to design around and significance of the patented feature to the product/service.

63.0/100 Score

Assignee Score

It takes the R&D behavior of the company itself into account that results in IP. During the invention phase, larger companies are considered to assign a higher R&D budget on a certain technology field, these companies have a better influence on their market, on what is marketable and what might lead to a standard.

17.0/100 Score

Legal Score

It shows the legal strength of IP in terms of its degree of protecting effect. Here we look into claim scope, claim breadth, claim quality, stability and priority.

Citation

Patents Cited in This Cited by
Title Current Assignee Application Date Publication Date
Microrna-based methods and compositions for reprogramming cells ALCEDO BIOTECH GMBH,BULLERDIEK, JÖRN,RIPPE, VOLKHARD 31 August 2010 03 March 2011
Viral and viral associated miRNAs and uses thereof ROSETTA GENOMICS LTD. 28 August 2006 14 September 2010
See full citation <>

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