US20040203020A1

US20040203020A1 - Self-replicating RNA molecule from hepatitis C virus

Info

Publication number: US20040203020A1
Application number: US10/686,835
Authority: US
Inventors: George Kukolj; Arnim Pause
Original assignee: Boehringer Ingelheim Canada Ltd
Current assignee: Boehringer Ingelheim Canada Ltd
Priority date: 2000-12-22
Filing date: 2003-10-16
Publication date: 2004-10-14
Also published as: DE60128835D1; US20040180333A1; EP1379660B1; WO2002052015A3; US20060246424A1; DE60128835T2; IL156058A0; WO2002052015A2; CA2430607C; JP2004516039A; JP2006304803A; ES2287074T3; AU2002218906B2; HUP0302593A3; US20020142350A1; HUP0302593A2; HU229567B1; EP1379660A2; US7344723B2; NZ527035A

Abstract

A unique HCV RNA molecule is provided having an enhanced efficiency of establishing cell culture replication. Novel adaptive mutations have been identified within the HCV non-structural region that improves the efficiency of establishing persistently replicating HCV RNA in cell culture. This self-replicating polynucleotide molecule contains, contrary to all previous reports, a 5′-NTR that can be either an A as an alternative to the G already disclosed and therefore provides an alternative to existing systems comprising a self-replicating HCV RNA molecule. The G-->A mutation gives rise to HCV RNA molecules that, in conjunction with mutations in the HCV non-structural region, such as the G(2042)C/R mutations, possess greater efficiency of transduction and/or replication. These RNA molecules when transfected in a cell line are useful for evaluating potential inhibitors of HCV replication.

Description

RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No. 10/029,907, filed Dec. 21, 2001, which claims, as does the present application priority to U.S. Provisional Application Serial No. 60/257,857 filed on Dec. 22, 2000, the disclosures of all of which are incorporated by reference in their entireties.[0001]

FIELD OF THE INVENTION

The present invention relates generally to a HCV RNA molecule that self-replicates in appropriate cell lines, particularly to a self-replicating HCV RNA construct having an enhanced efficiency of establishing cell culture replication.

BACKGROUND OF THE INVENTION

Hepatitis C virus (HCV) is the major etiological agent of post-transfusion and community-acquired non-A non-B hepatitis worldwide. It is estimated that over 200 million people worldwide are infected by the virus. A high percentage of carriers become chronically infected and many progress to chronic liver disease, so called chronic hepatitis C. This group is in turn at high risk for serious liver disease such as liver cirrhosis, hepatocellular carcinoma and terminal liver disease leading to death. The mechanism by which HCV establishes viral persistence and causes a high rate of chronic liver disease has not been thoroughly elucidated. It is not known how HCV interacts with and evades the host immune system. In addition, the roles of cellular and humoral immune responses in protection against HCV infection and disease have yet to be established.

Various clinical studies have been conducted with the goal of identifying pharmaceutical compounds capable of effectively treating HCV infection in patients afflicted with chronic hepatitis C. These studies have involved the use of interferon-alpha, alone and in combination with other antiviral agents such as ribavirin. Such studies have shown that a substantial number of the participants do not respond to these therapies, and of those that do respond favorably, a large proportion were found to relapse after termination of treatment. To date there are no broadly effective antiviral compounds for treatment of HCV infection. HCV is an enveloped positive strand RNA virus in the Flaviviridae family. The single strand HCV RNA genome is of positive polarity and comprises one open reading frame (ORF) of approximately 9600 nucleotides in length, which encodes a linear polyprotein of approx. 3010 amino acids. In infected cells, this polyprotein is cleaved at multiple sites by cellular and viral proteases to produce structural and non-structural (NS) proteins. The structural proteins (C, E1, E2 and E2-p7) comprise polypeptides that constitute the virus particle (Hijikata, M. et al., 1991, Proc. Natl. Acad. Sci. USA. 88, 5547-5551; Grakoui et al., 1993(a), J. Virol. 67,1385-1395). The non-structural proteins (NS2, NS3, NS4A, NS4B, NS5A, NS5B) encode for enzymes or accessory factors that catalyze and regulate the replication of the HCV RNA genome. Processing of the structural proteins is catalyzed by host cell proteases (Hijikata, M. et al., 1991, Proc. Natl. Acad. Sci. USA. 88, 5547-5551). The generation of the mature non-structural proteins is catalyzed by two virally encoded proteases. The first is the NS⅔ zinc-dependent metalloprotease which auto-catalyses the release of the NS3 protein from the polyprotein. The released NS3 contains a N-terminal serine protease domain (Grakoui et al., 1993(b), Proc Natl Acad Sci USA, 90,10583-7; Hijikata, M. et al., 1993, J. Virol. 67, 4665-4675) and catalyzes the remaining cleavages from the polyprotein. The released NS4A protein has at least two roles. First, forming a stable complex with NS3 protein and assisting in the membrane localization of the NS3/NS4A complex (Kim et al., Arch Virol. 1999, 144, 329-343) and second, acting as a cofactor for NS3 protease activity. This membrane-associated complex, in turn catalyzes the cleavage of the remaining sites on the polyprotein, thus effecting the release of NS4B, NS5A and NS5B (Bartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844; Grakoui et al., 1993(a), J. Virol. 67,1385-1395; Hijikata, M. et al., 1993, J. Virol. 67, 46654675; Love, R. A. et al., 1996, Cell, 87, 331-342; reviewed in Kwong et al., 1998 Antiviral Res., 40, 1-18). The C-terminal segment of the NS3 protein also harbors nucleoside triphosphatase and RNA helicase activity (Kim et al., 1995, Biochem. Biophys. Res. Comm., 215, 160-166.). The function of the protein NS4B is unknown. NS5A, a highly phosphorylated protein, seems to be responsible for the Interferon resistance of various HCV genotypes (Gale Jr. et al. 1997 Virology 230, 217; Reed et al., 1997 J. Virol. 71, 7187. NS5B is an RNA-dependent RNA polymerase (RdRp) that is involved in the replication of HCV.

The open reading frame of the HCV RNA genome is flanked on its 5′ end by a non-translated region (NTR) of approx. 340 nucleotides that functions as the internal ribosome entry site (IRES), and on its 3′ end by a NTR of approximately 230 nucleotides. Both the 5′ and 3′ NTRs are important for RNA genome replication. The genomic sequence variance is not evenly distributed over the genome and the 5′NTR and parts of the 3′NTR are the most highly conserved portions. The authentic, highly conserved 3′NTR is the object of U.S. Pat. No. 5,874,565 granted to Rice et al.

The cloned and characterized partial and complete sequences of the HCV genome have also been analyzed with regard to appropriate targets for a prospective antiviral therapy. Four viral enzyme activities provide possible targets such as (1) the NS2/3 protease; (2) the NS3/4A protease complex, (3) the NS3 Helicase and (4) the NS5B RNA-dependent RNA polymerase. The NS3/4A protease complex and the NS3 helicase have already been crystallized and their three-dimensional structure determined (Kim et al., 1996, Cell, 87, 343; Yem et al. Protein Science, 7, 837,1998; Love, R. A. et al., 1996, Cell, 87, 331-342; Kim et al., 1998, Structure, 6, 89; Yao et al., 1997 Nature Structural Biology, 4, 463; Cho et al., 1998, J. Biol. Chem., 273, 15045). The NS5B RNA dependent RNA polymerase has also been crystallized to reveal a structure reminiscent of other nucleic acid polymerases (Bressanelli et al. 1999, Proc. Natl. Acad. Sci, USA 96,13034-13039; Ago et al. 1999, Structure 7, 1417-1426; Lesburg et al. 1999, Nat. Struct. Biol. 6, 937-943).

Even though important targets for the development of a therapy for chronic HCV infection have been defined with these enzymes and even though a worldwide intensive search for suitable inhibitors is ongoing with the aid of rational drug design and HTS, the development of therapy has one major deficiency, namely the lack of cell culture systems or simple animal models, which allow direct and reliable propagation of HCV viruses. The lack of an efficient cell culture system is still the main reason to date that an understanding of HCV replication remains elusive. Although flavi- and pestivirus self-replicating RNAs have been described and used for the replication in different cell lines with a relatively high yield, similar experiments with HCV have not been successful to date (Khromykh et al., 1997, J. Virol. 71, 1497; Behrens et al., 1998, J. Virol. 72, 2364; Moser et al., 1998 J. Virol. 72, 5318). It is known from different publications that cell lines or primary cell cultures can be infected with high-titer patient serum containing HCV (Lanford et al. 1994 Virology 202, 606; Shimizu et al. 1993 PNAS, USA 90, 6037-6041; Mizutani et al. 1996 J. Virol. 70, 7219-7223; lkda, et al. 1998, Virus Res. 56,157; Fourner et al. 1998, J. Gen. Virol. 79, 2376; Ito et al. 1996, J. Gen. Virol. 77, 1043-1054). However, these virus-infected cell lines or cell cultures do not allow the direct detection of HCV-RNA or HCV antigens.

It is also known from the publications of Yoo et al. 1995 J. Virol., 69, 32-38; and of Dash et al., 1997, Am. J. Pathol., 151, 363-373; that hepatoma cell lines can be transfected with synthetic HCV-RNA obtained through in vitro transcription of the cloned HCV genome. In both publications the authors started from the basic idea that the viral HCV genome is a plus-strand RNA functioning directly as mRNA after being transfected into the cell, permitting the synthesis of viral proteins in the course of the translation process, and so new HCV particles could form HCV viruses and their RNA detected through RT-PCR. However the published results of the RT-PCR experiments indicate that the HCV replication in the described HCV transfected hepatoma cells is not particularly efficient and not sufficient to measure the quality of replication, let alone measure the modulations in replication after exposure to potential antiviral drugs. Furthermore it is now known that the highly conserved 3′ NTR is essential for the virus replication (Yanagi et al., 1999 Proc. Natl. Acad. Sci. USA, 96, 2291-95). This knowledge strictly contradicts the statements of Yoo et al. J. Virol., 69, 32-38(supra) and Dash et al., 1997, Am. J. Pathol., 151, 363-373. (supra), who used for their experiments only HCV genomes with shorter 3′ NTRs and not the authentic 3′ end of the HCV genome.

In WO 98/39031, Rice et al. disclosed authentic HCV genome RNA sequences, in particular containing: a) the highly conserved 5′-terminal sequence “GCCAGCC”; b) the HCV polyprotein coding region; and c) 3′-NTR authentic sequences.

In WO 99/04008, Purcell et al. disclosed an HCV infectious clone that also contained only the highly conserved 5′-terminal sequence “GCCAGC”.

Recently Lohman et al. 1999 (Science 285,110-113) and Bartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844(in CA 2,303,526, laid-open on Oct. 3, 2000) disclosed a HCV cell culture system where the viral RNA (1377/NS2-3′) self-replicates in the transfected cells with such efficiency that the quality of replication can be measured with accuracy and reproducibility. The Lohman and Bartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844 disclosures were the first demonstration of HCV RNA replication in cell culture that was substantiated through direct measurement by Northern blots. This replicon system and sequences disclosed therein highlight once again the conserved 5′ sequence “GCCAGC”. A similar observation highlighting the conservation of the 5′NTR was made by Blight et al. 2000 (Science 290,1972-1974) and WO 01/89364 published on Nov. 29, 2001.

In addition to the conservation of the 5′ and 3′ untranslated regions in cell culture replicating RNAs, three other publications by Lohman et al. 2001, J. Virol. 1437-1449 Krieger et al. 2001 J. Virol. 46144624 and Guo et al., (2001) J. Virol. 8516-8523 have recently disclosed distinct adaptive mutants within the HCV non-structural protein coding region. Specific nucleotide changes that alter the amino acids of the HCV non-structural proteins are shown to enhance the efficiency of establishing stable replicating HCV subgenomic replicons in culture cells.

Applicant has now found that, contrary to all previous reports, the highly conserved 5′-NTR can be mutated by adaptation to give rise to a HCV RNA sequence that, in conjunction with mutations in the HCV non-structural region, provides for a greater efficiency of transduction and/or replication.

Applicant has also identified novel adaptive mutations within the HCV non-structural region that improves the efficiency of establishing persistently replicating HCV RNA in cell culture.

One advantage of the present invention is to provide an alternative to these existing systems comprising a HCV RNA molecule that self-replicates. Moreover, the present invention demonstrates that the initiating nucleotide of the plus-strand genome can be either an A as an alternative to the G already disclosed.

A further advantage of the present invention is to provide a unique HCV RNA molecule that transduces and/or replicates with higher efficiency. The Applicant demonstrates the utility of this specific RNA molecule in a cell line and its use in evaluating a specific inhibitor of HCV replication.

SUMMARY OF THE INVENTION

In a first embodiment, the present invention provides a 5′-non translated region of the hepatitis C virus wherein its highly conserved guanine at position 1 is substituted for adenine.

Particularly, the present invention provides a hepatitis C virus polynucleotide comprising adenine at position 1 as numbered according to the 1377/NS2-3′ construct (Lohmann et al. 1999, Science 285,110-113, Accession # AJ242651).

Particularly, the invention provides a HCV self-replicating polynucleotide comprising a 5′-terminus consisting of ACCAGC (SEQ ID NO. 8).

In a second embodiment, the present invention is directed to a HCV self-replicating polynucleotide encoding a polyprotein comprising one or more amino acid substitution selected from the group consisting of: R(1 35)K; S(1148)G; S(1560)G; K(1691)R; L(1701)F; 1(1984)V; T(1993)A; G(2042)C; G(2042)R; S(2404)P; L(2155)P; P(2166)L and M(2992)T.

Particularly, the invention is directed to a HCV self-replicating polynucleotide encoding a polyprotein comprising the any one of the amino acid substitutions as described above, further comprising the amino acid substitution E(1202)G.

More particularly, the invention provides a HCV self-replicating polynucleotide encoding a polyprotein comprising a G2042C or a G2042R mutation.

Most particularly, the invention provides for HCV self-replicating polynucleotide comprising a nucleotide substitution G-->A at position 1, and said polynucleotide encodes a polyprotein further comprising a G2042C or a G2042R mutation.

Particularly, the polynucleotide of the present invention can be in the form of RNA or DNA that can be transcribed to RNA.

In a third embodiment, the invention also provides for an expression vector comprising a DNA form of the above polynucleotide, operably linked with a promoter.

According to a fourth embodiment, there is provided a host cell transfected with the self-replicating polynucleotide or the vector as described above.

In a fifth embodiment, the present invention provides a RNA replication assay comprising the steps of:

incubating the host cell as described above in the absence or presence of a potential hepatitis C virus inhibitor;

isolating the total cellular RNA from the cells;

analyzing the RNA so as to measure the amount of HCV RNA replicated;

comparing the levels of HCV RNA in cells in the absence and presence of the inhibitor.

In a sixth embodiment, the invention is directed to a method for testing a compound for inhibiting HCV replication, including the steps of:

a) treating the above described host cell with the compound;

b) evaluating the treated host cell for reduced replication, wherein reduced replication indicates the ability of the compound to inhibit replication.

DETAILED DESCRIPTION OF THE DRAWINGS

FIG. 1 is a schematic view of the bi-cistronic replicon RNA. The sequence deviations between the 1377/NS2-3′ replicon from Lohman et al., 1999 Science 285: 110-113 and the APGK12 replicon are indicated below the replicon. In place of a G nucleotide at the +1 position in the 1377/NS2-3′replicon, the APGK12 contains an additional G resulting in GG at the 5′ terminus (the first G being counted as position −1). In the linker region between the neo gene and the EMCV IRES sequence two areas deviate from 1377/NS2-3′: 14 nucleotides (CGCGCCCAGATGTT) which are not present in 1377/NS2/3 are inserted at position 1184 in APGK12; 11 nucleotides (1231-1241) present in 1377/NS2-3′ are deleted to generate APGK-12. In the NS5B coding region, a T at [0035] position 8032 was mutated to C to eliminate a NcoI restriction site.
FIG. 2 shows Northern blots of RNA-transfected Huh-7 cell lines. 12 μg of total cellular RNA or control RNA was separated on 0.5% agarose-formaldehyde gels and transferred to Hybond N+paper, fixed and (FIG. 2A) radioactively probed with HCV specific minus-strand RNA that detects the presence of plus-strand replicon RNA. [0036] Lanes 1 and 2: positive controls that contain 10⁹copies of in vitro transcribed APGK12 RNA. Lane 3: negative control of total cellular RNA from untransfected Huh-7 cells. Lanes 4 and 5: cellular RNA from B1 and B3 cell lines that have integrated DNA copies of the neomycin phosphotransferase gene. Lane 6: total cellular RNA from a Huh-7 cell line, designated S22.3, that harbors high copy number HCV sub-genomic replicon RNA as highlighted by the arrow. Other cell lines have no detectable replicon RNA. FIG. 2B is identical to FIG. 2A with the exception that the blot was radioactively probed with HCV specific plus-strand RNA to detect the presence of HCV minus-strand RNA. Lanes 1 and 2 are positive control lanes that contain 109 copies of full length HCV minus strand RNA. Lane 6, which contains 12 μg of total cellular RNA from cell line S22.3, harbors detectable minus-strand replicon RNA at the expected size of 8-9 kilobases. M represent the migration of non-radioactive molecular size markers on the agarose gel. 28s represents the migration of 28s ribosomal RNA and accounts for the detection of this species in a samples of total cellular RNA.
FIG. 3 shows indirect immunofluorescence of a HCV non-structural protein in the S22.3 cell line. Indirect immunofluorescence was performed on cells that were cultured and fixed, permeabilized and exposed to a rabbit polyclonal antibody specific for a segment of the HCV NS4A protein. Secondary goat anti-rabbit antibody conjugated with red-fluor Alexa 594 (Molecular Probes) was used for detection. Top panels shows the results of immunofluorescence (40× objective) and the specific staining of the S22.3 cells. The bottom panels represent the identical field of cells viewed by diffractive interference contrast (DIC) microscopy. The majority of S22.3 (FIG. 3A) cells within the field stain positively for HCV NS4A protein that localizes in the cytoplasm, whereas the B1 cells (FIG. 3B) that fail to express any HCV proteins, only have background level of staining. [0037]
FIG. 4 shows Western-blots following SDS-PAGE separation of total proteins extracted from three cell lines: (i) naive Huh-7 cell line, (ii) neomycin resistant Huh-7 cell line B1, and (iii) the S22.3 cell line. Panels A, B, and C, demonstrate the results of western blots probed with rabbit polyclonal antisera specific for neomycin phosphotransferase (NPT), HCV NS3, and HCV NS5B, respectively. Visualization was achieved through autoradiographic detection of a chemiluminescent reactive secondary\goat anti-rabbit antibody. Panel A shows that the S22.3 RNA replicon cell line, expresses the NPT protein at levels higher than control B1 cells and that the naïve Huh-7 cell line does not produce the NPT protein. Panels B and C show that only the S22.3 cell line produces the mature HCV NS3 and NS5B proteins, respectively. M represents molecular weight (in kilodaltons) of pre-stained polypeptide markers. [0038]
FIGS. 5A and 5B identify the nucleotide and amino acid sequences respectively that differ from the APGK12 sequence in the different HCV bi-cistronic replicons. The S22.3 adapted replicon is a first generation replicon selected following the transfection of RNA transcribed from the APGK12 template. R3, R7, R16 are second generation replicons that were selected following the transfection of RNA isolated from the S22.3 first generation replicon cell line. FIG. 5A: Nucleotide mutations that were characterized in each of the adapted replicons are indicated adjacent to the respective segment of the replicon (IRES, NS3, NS4A, NS5A, and NS5B). FIG. 5B: Amino acid numbers are numbered according to the full length HCV poly-protein with the first amino acid in the second cistron corresponding to [0039] amino acid 810 in NS2 of 1377/NS2-3′ construct.
FIG. 6 depicts the colony formation efficiency of four in vitro transcribed HCV sub-genomic bi-cistronic replicon RNAs. The APGK12 serves as the reference sequence; highlighted are the initiating nucleotides of the HCV IRES in each of the constructs and the amino acid differences (from the [0040] APGK1 2 reference sequence) in the HCV non-structural region for the two R3-rep. Note that the in vitro transcribed APGK-12 RNAs that harbor either a 5′G or 5′A form colonies with the same efficiency (ca. 80 cfu/μg in panels A and B) following selection with 0.25 mg/ml G418. RNA isolated from the second generation R3 cell line was reverse transcribed into DNA and cloned into the pAPGK12 vector backbone to generate the R3-rep, which was sequenced and found to encode additional changes that included the L(2155)P substitution in the NS5A segment of the HCV polyprotein (compare R3-rep sequence with the R3 sequence in tables 2 and 3). Various quantities of in vitro transcribed R3-rep-5′A RNA, were transfected into naïve Huh-7 cells to determine a colony formation efficiency of 1.2×10⁶cfu/μg of RNA (panel C). Various quantities of R3-rep-5′G were also transfected resulting in a colony formation efficiency of 2×10⁶cfu/μg of RNA (panel D).
FIG. 7 displays a typical RT-PCR amplification plot (left panel) and the graphical representation of Ct values versus known HCV RNA quantity in a standard curve (right panel). Each of the plotted curves in the left panel, graph the increment of fluorescence reporter signal (delta-Rn) versus PCR cycle number for a predetermined quantity of HCV replicon RNA. The Ct value is obtained by determining the point at which the fluorescence exceeds an arbitrary value (horizontal line). The right panel demonstrates the linear relationship between starting RNA copy number of the predetermined standards (large black dots) and the Ct value. Smaller dots are the Ct values of RNA samples (containing unknown quantity of HCV replicon RNA) from S22.3 cells treated with various concentrations of a specific inhibitor of HCV replication. [0041]
FIG. 8 shows the effect of increasing concentration of inhibitor A on HCV RNA replicon levels in Huh7 cells. S22.3 cells were grown in the presence of increasing concentrations of inhibitor A starting at 0.5 nM and ranging to 1024 nM. The inhibitor dose-response curve is the result of 11 concentrations from serial two-fold dilutions (1:1). One control well, without any inhibitor, was also included during the course of the experiment. The cells were incubated for 4 days in a 5% CO[0042] ₂incubator at 37° C. Total cellular RNA was extracted, quantified by optical density. HCV replicon RNA was evaluated by real time RT-PCR and plotted as genome equivalents/μg total RNA as a function of inhibitor concentration

DEFINITIONS

Unless defined otherwise, the scientific and technological terms and nomenclature used herein have the same meaning as commonly understood by a person of ordinary skill to which this invention pertains. Generally, the procedures for cell culture, infection, molecular biology methods and the like are common methods used in the art. Such standard techniques can be found in reference manuals such as for example Sambrook et al. (1989) Molecular Cloning—A Laboratory Manual, Cold Spring Harbor Labs and Ausubel et al. (1994). Nucleotide sequences are presented herein by single strand, in the 5′ to 3′ direction, from left to right, using the one letter nucleotide symbols as commonly used in the art and in accordance with the recommendations of the IUPAC-IUB Biochemical [0043]
Nomenclature Commission (1972) Biochemistry, 11, 1726-1732. The present description refers to a number of routinely used recombinant DNA (rDNA) technology terms. Nevertheless, definitions of selected examples of such rDNA terms are provided for clarity and consistency. [0044]
The term “DNA segment or molecule or sequence”, is used herein, to refer to molecules comprised of the deoxyribonucleotides adenine (A), guanine (G), thymine (T) and/or cytosine (C). These segments, molecules or sequences can be found in nature or synthetically derived. When read in accordance with the genetic code, these sequences can encode a linear stretch or sequence of amino acids which can be referred to as a polypeptide, protein, protein fragment and the like. [0045]
As used herein, the term “gene” is well known in the art and relates to a nucleic acid sequence defining a single protein or polypeptide. The polypeptide can be encoded by a full-length sequence or any portion of the coding sequence, so long as the functional activity of the protein is retained. [0046]
A “structural gene” defines a DNA sequence which is transcribed into RNA and translated into a protein having a specific structural function that constitute the viral particles. “Structural proteins” defines the HCV proteins incorporated into the virus particles namely, core “C”, E1, E2, and E2-p7. [0047]
“Non-structural proteins”, defines the HCV proteins that are not comprised in viral particles namely, NS2, NS3, NS4A, NS5A and NS5B. [0048]
“Restriction endonuclease or restriction enzyme” is an enzyme that has the capacity to recognize a specific base sequence (usually 4, 5 or 6 base pairs in length) in a DNA molecule, and to cleave the DNA molecule at every place where this sequence appears. An example of such an enzyme is EcoRI, which recognizes the base sequence G⇓AATTC and cleaves a DNA molecule at this recognition site. [0049]
“Restriction fragments” are DNA molecules produced by the digestion of DNA with a restriction endonuclease. Any given genome or DNA segment can be digested by a particular restriction endonuclease into at least two discrete molecules of restriction fragments. [0050]
“Agarose gel electrophoresis” is an analytical method for fractionating polynucleotide molecules based on their size. The method is based on the fact that nucleic acid molecules migrate through a gel as through a sieve, whereby the smallest molecule has the greatest mobility and travels the farthest through the gel. The sieving characteristics of the gel retards the largest molecules such that, these have the least mobility. The fractionated polynucleotides can be visualized by staining the gel using methods well known in the art, nucleic acid hybridization or by tagging the fractionated molecules with a detectable label. All these methods are well known in the art, specific methods can be found in Ausubel et al. (supra). [0051]
“Oligonucleotide or oligomer” is a molecule comprised of two or more deoxyribonucleotides or ribonucleotides, preferably more than three. The exact size of the molecule will depend on many factors, which in turn depend on the ultimate function or use of the oligonucleotide. An oligonucleotide can be derived synthetically, by cloning or by amplification. [0052]
“Sequence amplification” is a method for generating large amounts of a target sequence. In general, one or more amplification primers are annealed to a nucleic acid sequence. Using appropriate enzymes, sequences found adjacent to, or in between the primers are amplified. An amplification method used herein is the polymerase chain reaction (PCR) and can be used in conjunction with the reverse-transcriptase (RT) to produce amplified DNA copies of specific RNA sequences. [0053]
“Amplification primer” refers to an oligonucleotide, capable of annealing to a RNA or DNA region adjacent to a target sequence and serving as the initiation primer for DNA synthesis under suitable conditions well known in the art. The synthesized primer extension product is complementary to the target sequence. [0054]
The term “domain” or “region” refers to a specific amino acid sequence that defines either a specific function or structure within a protein. As an example herein, is the NS3 protease domain comprised within the HCV non-structural polyprotein. [0055]
The terms “plasmid” “vector” or “DNA construct” are commonly known in the art and refer to any genetic element, including, but not limited to, plasmid DNA, phage DNA, viral DNA and the like which can incorporate the oligonucleotide sequences, or sequences of the present invention and serve as DNA vehicle into which DNA of the present invention can be cloned. Numerous types of vectors exist and are well known in the art. [0056]
The terminology “expression vector” defines a vector as described above but designed to enable the expression of an inserted sequence following transformation or transfection into a host. The cloned gene (inserted sequence) is usually placed under the control of control element sequences such as promoter sequences. Such expression control sequences will vary depending on whether the vector is designed to express the operably linked gene in vitro or in vivo in a prokaryotic or eukaryotic host or both (shuttle vectors) and can additionally contain transcriptional elements such as enhancer elements, termination sequences, tissue-specificity elements, and/or translational initiation and termination sites. [0057]
A host cell or indicator cell has been “transfected” by exogenous or heterologous DNA (e.g. a DNA construct) or RNA, when such nucleic acid has been introduced inside the cell. The transfecting DNA may or may not be integrated (covalently linked) into chromosomal DNA making up the genome of the cell. In prokaryotes, yeast, and mammalian cells for example, the transfecting/transforming DNA may be maintained on an episomal element such as a plasmid. With respect to eukaryotic cells, an example of a stably transfected cell is one in which the transfecting DNA has become integrated into a chromosome and is inherited by daughter cells through chromosome replication. A host cell or indicator cell can be transfected with RNA. A cell can be stably transfected with RNA if the RNA replicates and copies of the RNA segregate to daughter cells upon cell division. This stability is demonstrated by the ability of the eukaryotic cell to establish cell lines or clones comprised of a population of daughter cells containing the transfecting DNA or RNA. Transfection methods are well known in the art (Sambrook et al., 1989, Molecular Cloning—A Laboratory Manual, Cold Spring Harbor Labs; Ausubel et al., 1994, Current Protocols in Molecular Biology, Wiley, New York). If the RNA encodes for a genetic marker that imparts an observable phenotype, such as antibiotic resistance, then the stable transfection of replicating RNA can be monitored by the acquisition of such phenotype by the host cell. [0058]
As used herein the term “transduction” refers to the transfer of a genetic marker to host cells by the stable transfection of a replicating RNA. [0059]
The nucleotide sequences and polypeptides useful to practice the invention include without being limited thereto, mutants, homologs, subtypes, quasi-species, alleles, and the like. It is understood that generally, the sequences of the present invention encode a polyprotein. It will be clear to a person skilled in the art that the polyprotein of the present invention and any variant, derivative or fragment thereof, is auto-processed to an active protease. [0060]
As used herein, the designation “variant” denotes in the context of this invention a sequence whether a nucleic acid or amino acid, a molecule that retains a biological activity (either functional or structural) that is substantially similar to that of the original sequence. This variant may be from the same or different species and may be a natural variant or be prepared synthetically. Such variants include amino acid sequences having substitutions, deletions, or additions of one or more amino acids, provided the biological activity of the protein is conserved. The same applies to variants of nucleic acid sequences which can have substitutions, deletions, or additions of one or more nucleotides, provided that the biological activity of the sequence is generally maintained. [0061]
The term “derivative” is intended to include any of the above described variants when comprising additional chemical moiety not normally a part of these molecules. These chemical moieties can have varying purposes including, improving a molecule's solubility, absorption, biological half life, decreasing toxicity and eliminating or decreasing undesirable side effects. Furthermore, these moieties can be used for the purpose of labeling, binding, or they may be comprised in fusion product(s). Different moieties capable of mediating the above described effects can be found in Remington's The Science and Practice of Pharmacy (1995). Methodologies for coupling such moieties to a molecule are well known in the art. The term “fragment” refers to any segment of an identified DNA, RNA or amino acid sequence and/or any segment of any of the variants or derivatives described herein above that substantially retains its biological activity (functional or structural) as required by the present invention. [0062]
The terms “variant”, “derivative”, and “fragment” of the present invention refer herein to proteins or nucleic acid molecules which can be isolated/purified, synthesized chemically or produced through recombinant DNA technology. All these methods are well known in the art. As exemplified herein below, the nucleotide sequences and polypeptides used in the present invention can be modified, for example by in vitro mutagenesis. [0063]
As used herein, the term “HCV polyprotein coding region” means the portion of a hepatitis C virus that codes for the polyprotein open reading frame (ORF). This ORF may encode proteins that are the same or different than wild-type HCV proteins. The ORF may also encode only some of the functional protein encoded by wild-type polyprotein coding region. The protein encoded therein may also be from different isolates of HCV, and non-HCV protein may also be encoded therein. [0064]
As used herein, the abbreviation “NTR” used in the context of a polynucleotide molecule means a non-translated region. The term “UTR” means untranslated region. Both are used interchangeably. [0065]
Preferred Embodiments [0066]
Particularly, the invention provides a HCV self-replicating polynucleotide molecule comprising a 5′-terminus consisting of ACCAGC (SEQ ID NO.8). [0067]
According to the first embodiment of this invention, there is particularly provided a HCV polynucleotide construct comprising: [0068]
a 5′-non translated region (NTR) comprising the sequence ACCAGC at, or proximal to, its 5′-terminus; [0069]
a HCV polyprotein coding region; and [0070]
a 3′-NTR region. [0071]
In a second embodiment, the present invention is directed to a HCV self-replicating polynucleotide encoding a polyprotein comprising one or more amino acid substitution selected from the group consisting of: R(1135)K; S(1148)G; S(1560)G; K(1691)R; L(1701)F; 1(1984)V; T(1993)A; G(2042)C; G(2042)R; S(2404)P; L(2155)P; P(2166)L and M(2992)T. [0072]
Particularly, the invention is directed to a HCV self-replicating polynucleotide encoding a polyprotein comprising the any one of the amino acid substitutions as described above, further comprising the amino acid substitution E(1202)G. [0073]
Alternatively, the first embodiment of the present invention is directed to HCV self-replicating polynucleotide molecule comprising a G2042C/R mutation. [0074]
According to the second embodiment, the present invention particularly provides a HCV polynucleotide construct comprising: [0075]
a 5′-NTR region comprising the sequence ACCAGC at, or proximal to, its 5′-terminus; [0076]
a HCV polyprotein region coding for a HCV polyprotein comprising a G(2042)C or a G(2042)R mutation; and [0077]
a 3′-NTR region. [0078]
Preferably, the polynucleotide construct of the present invention is a DNA or RNA molecule. More preferably, the construct is a RNA molecule. Most preferably, the construct is a DNA molecule. [0079]
More particularly, the first embodiment of this invention is directed to a RNA molecule encoded by the DNA molecule selected from the group consisting of: SEQ ID NO.2, 4, 5, 6, 7, 24 and 25. [0080]
Most particularly, the invention provides a DNA molecule selected from the group consisting of: SEQ ID NO. 2, 4, 5, 6, 7, 24 and 25. [0081]
In a third embodiment, the invention also is directed to an expression vector comprising DNA forms of the above polynucleotide, operably linked with a promoter. [0082]
Preferably, the promoter is selected from the group consisting of: T3, T7 and SP6. [0083]
According to a fourth embodiment, there is provided a host cell transfected with the self-replicating polynucleotide or vector as described above. Particularly, the host cell is a eukaryotic cell line. More particularly, the eukaryotic cell line is a hepatic cell line. Most particularly, the hepatic cell line is Huh-7. [0084]
In a fifth embodiment, the present invention provides a RNA replication assay comprising the steps of: [0085]
a) incubating the host cell as described above under conditions suitable for RNA replication; [0086]
b) isolating the total cellular RNA from the cells; and [0087]
c) analyzing the RNA so as to measure the amount of HCV RNA replicated. [0088]
Preferably, the analysis of RNA levels in step c) is carried out by amplifying the RNA by real-time RT-PCR analysis using HCV specific primers so as to measure the amount of HCV RNA replicated. [0089]
Alternatively in this fifth embodiment, the construct comprises a reporter gene, and the analysis of RNA levels in step c) is carried out by assessing the level of reporter expressed. [0090]
According to a preferred aspect of the sixth embodiment, the invention is directed to a method for testing a compound for inhibiting HCV replication, including the steps of: [0091]
a) carrying step a) as described in the above assay, in the presence or absence of the compound; b) isolating the total cellular RNA from the cells; and [0092]
c) analyzing the RNA so as to measure the amount of HCV RNA replicated. [0093]
d) comparing the levels of HCV RNA in cells in the absence and presence of the inhibitor, [0094]
wherein reduced RNA levels is indicative of the ability of the compound to inhibit replication. [0095]
Preferably, the cell line is incubated with the test compound for about 34 days at a temperature of about 37° C. [0096]

EXAMPLES

EXAMPLE 1

Replicon Constructs (APGK-12; FIG. 1) [0097]
pET9a-EMCV was obtained by ligating an [0098] oligonucleotide linker 5′ gaattccagatggcgcgcccagatgttaaccagatccatggcacactctagagtactgtcgac 3′ (SEQ ID NO.9) to pET-9a (Novagen) that was cut with EcoRI and SalI to form the vector pET-9a-mod. This linker contains the following restriction sites: EcoRI, AscI, Hpal, NcoI, XbaI, ScaI, SaII. The EMCV IRES was amplified by PCR from the vector pTM1 with primers 5′ cggaatcgttaacagaccacaacggtttccctc 3′ (SEQ ID NO.10) and 5′ ggcgtacccatggtattatcgtgtttttca 3′ (SEQ ID NO.11) and ligated into pET-9a-mod via EcoRI and NcoI to form pET-9a-EMCV.
The sequence of HCV NS2 to NS5B followed by the 3′UTR of HCV was obtained from the [0099] replicon construct 1377/NS2-3′ (Lohman et al., 1999 Science 285:110-113; accession number: AJ242651) and synthesized by Operon Technologies Inc. with a T to C change at the NcoI site in NS5B at nucleotide 8032. This sequence was released from an GenOp® vector (Operon Technologies) with NcoI and ScaI and transferred into pET-9a-EMCV to form pET-9a-EMCV-NS2-5B-3′UTR.
pET-9a-HCV-neo was obtained by amplification of the HCV IRES from a HCV cDNA isolated from patient serum with [0100] primers 5′ gcatatgaattctaatacgactcactataggccagcccccgattg 3′ (SEQ ID NO.12) containing a T7 promoter and primer 5′ ggcgcgccctttggtttttctttgaggtttaggattcgtgctcat 3′ (SEQ ID NO.13) and amplification of the neomycin phosphotransferase gene from the vector pcDNA 3.1 (Invitrogen) with primers 5′ aaagggcgcatgattgaacaagatggattgcacgca 3′ (SEQ ID NO.14) and 5′ gcatatgttaactcagaagaactcgtcaagaaggcgata 3′ (SEQ ID NO.15). These two PCR fragments were mixed and amplified with primers 5′ gcatatgaattctaatacgactcactataggccagcccccgattg 3′ (SEQ ID NO.16) and 5′ gcatatgttaactcagaagaactcgtcaagaaggcgata 3′ (SEQ ID NO.15), cut with EcoRI and Hpal and transferred into pET-9a-mod to form pet-9a-HCV-neo. The EMCV-NS2-5B-3′UTR was released from pET-9a-EMCV-NS2-5B-3′UTR with Hpal and ScaI and transferred into pet-9a-HCV-neo that was cut with Hpal to form pET-9a- APGK12. This insert was sequenced with specific successive primers using a ABI Prism® BigDye™ Terminator Cycle sequencing kit and analyzed on ABI Prism® 377 DNA Sequencer and is shown in SEQ ID NO 1.
RNA In Vitro Transcription [0101]
pET-9a-APGK12 DNA was cut with ScaI for expression of the full-length replicon or with BgIII for expression of a truncated negative control RNA. DNA was analyzed on a 1% agarose gel and purified by Phenol/Chloroform extraction. RNA was produced using a T7 Ribomax® kit (Promega) followed by extraction with phenol/chloroform and precipitation with 7.5 M LiCl[0102] ₂. RNA was treated with DNAse I for 15 min to remove the DNA template and further purified with an RNeasy® column (Qiagen). RNA integrity was verified on a denaturing formaldehyde 1% agarose gel.

EXAMPLE 2

Primary transfection of Huh7 cells and selection of replicon cell lines Human hepatoma Huh7 cells (Health Science Research Resources Bank, Osaka, Japan) were grown in 10% FBS/DMEM. Cells were grown to 70% confluency, trypsinized, washed with phosphate buffered saline (PBS) and adjusted to 1×10[0103] ⁷cells/ml of PBS. 800 μl of cells were transferred into 0.4 cm cuvettes and mixed with 15 μg of replicon RNA. Cells were electroporated using 960μF, 300 volts for ˜18 msec and evenly distributed into two 15 cm tissue culture plates and incubated in a tissue culture incubator for 24 hours. The selection of first and second generation replicon cell lines was with 10% FBS/DMEM medium supplemented with 1 mg/ml of G418. Cells were selected for 3-5 weeks until colonies were observed that were isolated and expanded.
Following the G418 selection and propagation of Huh-7 cells transfected with APGK12 (SEQ ID NO. 1) RNA, cells that formed a distinct colony were treated with trypsin and serially passed into larger culture flasks to establish cell lines. Approximately 10×10[0104] ⁶cells were harvested from each cell line. The cells were lysed and the total cellular RNA extracted and purified as outlined in Qiagen RNAeasy® preparatory procedures. FIG. 2 shows the analysis of 12 μg of total cellular RNA from various cell lines as analyzed on a Northern blot of a denaturing agarose-formaldehyde gel.
FIG. 2A is a Northern blot (radioactively probed with HCV specific minus-strand RNA) that detects the presence of plus-strand replicon RNA. [0105] Lanes 1 and 2 are positive controls that contain 109 copies of in vitro transcribed APGK12 RNA. Lane 2 contains the in vitro transcribed RNA mixed with 12 μg of total cellular from naive Huh-7 cells. Lane 3 is a negative control of total cellular RNA from untreated Huh-7 cells. Lanes 4 and 5 contain cellular RNA from the B1 and B3 G418 resistant cell lines that have DNA integrated copies of the neomycin phosphotransferase gene. Lane 6 contains total cellular RNA from a Huh-7 cell line, designated S22.3, that harbors high copy number of HCV sub-genomic replicon RNA as detected by the positive signal in the 8 kilo-base range. Other cell lines have no detectable replicon RNA. FIG. 2B is a Northern blot of a duplicate of the gel presented in 2A with the exception that the blot was radioactively probed with HCV specific plus-strand RNA to detect the presence of HCV minus-strand RNA ( lanes 1 and 2 are positive control lanes that contain 10⁹copies of full length genomic HCV minus strand RNA); only lane 6, which contains 12 μg of total cellular RNA from cell line S22.3, harbors detectable minus-strand replicon RNA at the expected size of 8-9 kilobases. An quantitative estimation of RNA copy number, based on phosphorimager scanning of the Northern blots, is approximately 6×10⁷copies of plus-strand/μg of total RNA, and 6×10⁶copies of minus strand/μg of total RNA. The presence of the plus-strand and minus-strand intermediate confirms that the HCV sub-genomic RNA is actively replicating in the S22.3 cell line.

Example 3

S22.3 Cell Line Constitutively Expresses HCV Non-structural Proteins. [0106]
HCV non-structural protein expression was examined in the S22.3 cell line. FIG. 3 displays the result of indirect immunofluorescence that detects the HCV NS4A protein in the S22.3 cell line and not in the replicon negative B1 cell line (a G418 resistant Huh-7 cell line). Indirect immunofluorescence was performed on cells that were cultured and fixed (with 4% paraformaldehyde) onto Lab-tek chamber slides. Cells were permeabilized with 0.2% Triton X-100 for 10 minutes followed by a 1 hour treatment with 5% milk powder dissolved in phosphate-buffered saline (PBS). A rabbit serum containing polyclonal antibody raised against a peptide spanning the HCV NS4A region was the primary antibody used in detection. Following a 2 hour incubation with the primary antibody, cells were washed with PBS and a secondary goat anti-rabbit antibody conjugated with red-fluor Alexa® 594 (Molecular Probes) was added to cells for 3 hours. Unbound secondary antibody was removed with PBS washes and cells were sealed with a cover slip. FIG. 3 (top panels) shows the results of immunofluorescence as detected by a microscope with specific fluorescent filtering; the bottom panels represent the identical field of cells viewed by diffractive interference contrast (DIC) microscopy. The majority of S22.3 (FIG. 3A) cells within the field stain positively for HCV NS4A protein that localizes in the cytoplasm, whereas the B1 cells (FIG. 3B) that fail to express any HCV proteins, only have background level of staining. A small proportion of S22.3 cells express high levels of intensely stained HCV NS4A. [0107]
Expression of the proteins encoded by the bi-cistronic replicon RNA was also examined on Western-blots following SDS-PAGE separation of total proteins extracted from: (i) naive Huh-7 cell line, (ii) neomycin resistant Huh-7 cell line B1, and (iii) the S22.3 cell line. FIG. 4 panels A, B, and C, demonstrate the results of western blots probed with rabbit polyclonal antisera specific for neomycin phosphotransferase (NPT), HCV NS3, and HCV NS5B, respectively. Visualization was achieved through autoradiographic detection of a chemiluminescent reactive secondary HRP-conjugated goat anti-rabbit antibody. FIG. 4 panel A shows that the S22.3 RNA replicon cell line, expresses the NPT protein at levels higher than B1 cells (which contain an integrated DNA copy of the npt gene) and that the naïve Huh-7 cell line does not produce the NPT protein. FIG. 4 panels B and C show that only the S22.3 cell line produces the mature HCV NS3 and NS5B proteins, respectively. The western blots demonstrate that the S22.3 cell line, which harbors actively replicating HCV sub-genomic replicon RNA, maintains replication of the RNA through the high level expression of the HCV non-structural proteins. [0108]

Example 4

Sequence Determination of Adapted Replicons [0109]
Total RNA was extracted from replicon containing Huh7 cells using a RNeasy Kit (Qiagen). Replicon RNA was reverse transcribed and amplified by PCR using a OneStep RT-PCR kit (Qiagen) and HCV specific primers (as selected from the full-length sequence disclosed in WO 00/66623). Ten distinct RT-PCR products, that covered the entire bi-cistronic replicon in a staggered fashion, were amplified using oligonucleotide primers. The PCR fragments were sequenced directly with ABI Prism® BigDye™ Terminator Cycle PCR Sequencing and analyzed on ABI Prism® 377 DNA Sequencer. To analyze the sequence of the [0110] HCV replicon 3′ and 5′ ends a RNA ligation/RT-PCR procedure described in Kolykhalov et al. 1996 J. of Virology, 7, p. 3363-3371 was followed. The nucleotide sequence of S22.3 is presented as SEQ ID NO. 2.

Example 5

Serial Passage of HCV Replicon RNA [0111]

The total cellular RNA from the S22.3 cell line was prepared as described above. HCV Replicon RNA copy number was determined by Taqman® RT-PCR analysis and 20 μg of total S22.3 cellular RNA (containing 1×10 ⁹copies of HCV RNA) was transfected by electroporation into 8×10⁶naive Huh-7 cells. Transfected cells were subsequently cultured in 10 cm tissue culture plates containing DMEM supplemented with 10% fetal calf serum (10% FCS). Media was changed to DMEM (10% FCS) supplemented with 1 mg/ml G418 24 hours after transfection and then changed every three days. Twenty-three visible colonies formed three to four weeks post-transfection and G418 selection. G418 resistant colonies were expanded into second generation cell lines that represent the first cell lines harboring serially passaged HCV Replicon RNA. Three of these cell lines: R3, R7, and R16 were the subject of further analyses. First, the efficiency of transduction by each of the adapted replicons was determined by electroporation of the total cellular RNA (extracted from the R3, R7 and R16) into naive Huh-7 cells; following electroporation, the transduction efficiency was determined as described above, by counting the visible G418 resistant colonies that arose following 3 to 5 weeks of G418 selection (Table 1). Second, the sequence of the serially passed adapted replicons was determined from the total cellular RNA that was extracted from each of the R3, R7 and R16 replicon cell lines as described in example 4 (SEQ ID NO. 4, 5, 6). Using the pAPGK12 as a reference sequence (SEQ ID NO. 1), the nucleotide changes that were selected in HCV segment of the adapted replicons are presented in FIG. 5A. Some of these nucleotide changes are silent and do not change the encoded amino acid whereas others result in an amino acid substitution. FIG. 5B summarizes the amino acid changes encoded by the adapted replicons with the amino acid sequence of pAPGK12 as the reference. It is important to note that the reference sequence APGK-12 (SEQ ID NO.1) contains an extra G at the 5′-terminal (5′-GG) that is not maintained in the replicating RNA of the established cell lines. Also noteworthy is that, in addition to G->A at nucleotide 1, there is also an adapted mutation G->C/R at amino acid 2042 (shown as amino acid 1233 in the sequence listing since a.a. 810 of NS2 is numbered as a.a. 1 in SEQ ID) that can be found in all clones analyzed.

TABLE 1


Transfection of Huh-7 cells

RNA	Copies of Replicon	# Colonies	SEQ ID

5 ng APKG12 replicon	1.2 × 10⁹	0
in 20 μg total Huh-7 RNA
15 μg APKG12	3 × 10¹²	1 (S22.3)	1
replicon RNA
20 μg total:	3 × 10⁹	23 (3 clones	2
S22.3 cellular RNA		analyzed)
R3 cellular RNA	1 × 10⁹	200	4
R7 cellular RNA	1 × 10⁹	20	5
R16 cellular RNA	3 × 10⁸	100	6
cloned R3rep RNA	2.3 × 10⁸	2000	7

Example 6

Construction of APGK12 with 5′ G->A substitution (APGK12-5′A, SEQ ID NO.24) The pAPGK12 DNA was modified to change the first nucleotide in the sequence to replace the 5′GG with a 5′A. The change in the pAPGK12 was introduced by replacing an EcoRI/AgeI portion of the sequence with a PCR-generated EcoRI/AgeI fragment that includes the mutation. The oligonucleotides used for the amplification were (SEQ ID. NO. 20): 5′-GTG GAC GAA TTC TAA TAC GAC TCA CTA TA[0113] A CCA GCC CCC GAT TGG-3′ and (SEQ ID. NO. 21): 5′-GGA ACG CCC GTC GTG GCC AGC CAC GAT-3′ and generated a 195 bp DNA fragment that was then digested with EcoRI and AgeI. The resulting 178 bp restriction fragment was used to replace the EcoRI/AgeI fragment in pAPGK12 to generate the pAPGK12-5′A plasmid.

Example 7

cDNA Cloning Of The R3-replicon (R3REP). [0114]
The cDNA clone of the R3 replicon was produced by RT-PCR of RNA extracted from the R3 cell line. The following two oligonucleotides were used: (SEQ ID. NO. 22): 5′-GTC GTC TTC TCT GAC ATG GAG AC-3′ and (SEQ ID. NO. 23): 5′-GAG TTG CTC AGT GGA TTG ATG GGC AGC-3′. The ˜4400 nt PCR fragment, starting within the NS2 coding region and extending to the 5′-end of the NS5B coding region, was cloned into the plasmid pCR3.1 by TA cloning (Invitrogen). The SacII/XhoI portion of this R3 sequence was then used to replace the SacII/XhoI fragment present in the pAPGK12 and the pAPGK12-5′A described above. Consequently, two R3 cDNA sequences were generated: (I) R3-Rep-5′G with an initiating 5′G (SEQ ID NO.7), and R3-Rep-5′A (SEQ ID NO.25) with an initiating 5′A. Sequencing of the R3 rep cDNA identified unique nucleotide changes that differ from the original pAPGK12 sequence (see FIG. 5A); some of these changes are silent and do not change the encoded amino acid, whereas others do result in an amino acid change (see FIG. 5B). The differences between R3 and the R3-rep reflect the isolation of a unique R3-rep cDNA clone encoding nucleotide changes that were not observed from the sequencing of the total RNA extracted from the R3 cell line. [0115]

Example 8

Efficiency of Colony Formation With Modified Constructs [0116]
RNA from pAPGK12, pAPGK12-5′A, pR3-Rep and pR3-Rep-5′A was generated by in vitro transcription using the T7 Ribomax® kit (Promega) as described in example 1 above. The reactions containing the pAPGK12-5′A and pR3-Rep-5′A templates were scaled-up 10-fold due to the limitation of commercial RNA polymerase in initiating transcripts with 5′-A. The full length RNAs and control truncated RNA for each clone were introduced into 8×10[0117] ⁶naive Huh-7 cells by electroporation as described in example 2. Replicon RNA was supplemented with total cellular Huh-7 carrier RNA to achieve a final 15-20 μg quantity. The cells were then cultured in DMEM medium supplemented with 10% fetal calf serum and 0.25 mg/ml G418 in two 150 mm plates. The lower concentration of G418 was sufficient to isolate and select replicon containing cell lines as none of the transfectants with the control truncated RNA produced any resistant colonies. In contrast, in vitro transcribed APGK-12 RNAs that harbor either a 5′G or 5′A form colonies with the same efficiency (ca. 80 cfu/μg in FIG. 6 panels A and B) following selection with G418. Various quantities (ranging from 0.1 ng to 1 μg) of the R3-rep-5′A RNA, were transfected into naïve Huh-7 cells to determine a colony formation efficiency of 1.2×10⁶cfu/μg of RNA (FIG. 6 panel C depicts transfection with 1 μg of RNA). Various quantities (ranging from 0.1 ng to 1 μg) of R3-rep [5′G] were similarly transfected resulting in a colony formation efficiency of 2×10⁶cfu/μg of RNA (FIG. 6 panel D depicts colony formation with 1 μg of RNA). Note that, shown for the first time, HCV subgenomic replicons replicate as efficiently with a 5′ A nucleotide in place of the 5′G. APGK12 with a 5′A or 5′G RNA have similar transduction efficiencies. Similarly, R3-Rep RNAs with either the 5′A or 5′G both display the markedly increased transduction efficiency. Notably, the adaptive mutants within the HCV non-structural segment encoded by the R3-Rep provides for a substantial increase in transduction efficiency as depicted by the dramatic increase in colony forming units per μg of transfected RNA.

Example 9

Quantification of HCV Replicon RNA Levels in Cell Lines [0118]

S22.3 cells, or cell lines harboring other adapted replicons, were seeded in DMEM supplemented with 10% FBS, PenStrep and 1 μg/mL Geneticin. At the end of the incubation period the replicon copy number is evaluated by real-time RT-PCR with the ABI Prism 7700 Sequence Detection System. The TAQMAN® EZ RT-PCR kit provides a system for the detection and analysis of HCV RNA (as first demonstrated by Martell et al. 1999 J. Clin. Microbiol. 37: 327-332). Direct detection of the reverse transcription polymerase chain reaction (RT-PCR) product with no downstream processing is accomplished by monitoring the increase in fluorescence of a dye-labeled DNA probe (FIG. 6). The nucleotide sequence of both primers (adapted from Ruster, B. Zeuzem, S. and Roth, W. K., 1995. Analytical Biochemistry 224:597-600) and probe (adapted from Hohne, M., Roeske, H. and Schreier, E. 1998, Poster Presentation: P297 at the Fifth International Meeting on Hepatitis C Virus and Related Viruses Molecular Virology and Pathogenesis, Venezia-Lido Italy, June 25-28,1998) located in the 5′-region of the HCV genome are the following: HCV Forward primer:


HCV Forward primer:
5′ ACG CAG AAA GCG TCT AGC CAT GGC	(SEQ ID NO.17)

GTT AGT 3′

HCV Reverse primer:
5′ TCC CGG GGC ACT CGC AAG CAC CCT	(SEQ ID NO.18)

ATC AGG 3′

HCV Probe:
5′ FAM-TGG TCT GCG GAA CGG GTG AGT	(SEQ ID NO.19)

ACA CC-TAMRA 3′

FAM: Fluorescence reporter dye. [0120]
TAMRA: Quencher dye. [0121]

Using The TAQMAN® EZ RT-PCR kit, the following reaction was set up:



	Volume per sample
Component	(μL)	Final Concentration

RNase-Free Water	16	—
5 × Taqman EZ Buffer	10	1×
Manganese Acetate 25 mM	6	3 mM
dATP
10 mM	1.5	300 μM
dCTP
10 mM	1.5	300 μM
dGTP
10 mM	1.5	300 μM
dUTP
20 mM	1.5	300 μM
HCV Forward Primer 10 μM	1	200 nM
HCV Reverse Primer 10 μM	1	200 nM
HCV Probe
5 uM	2	200 nM
rTth DNA Polymerase	2	0.1 U/μL
2.5 U/μL
AmpErase UNG 1 U/μL	0.5	0.01 U/μL
Total Mix	45	—

To this reaction mix, 5 μL of total RNA extracted from S22.3 cells diluted at 10 ng/μL was added, for a total of 50 ng of RNA per reaction. The replicon copy number was evaluated with a standard curve made from known amounts of replicon copies (supplemented with 50 ng of wild type Huh-7 RNA) and assayed in an identical reaction mix (FIG. 7). [0123]

Thermal cycler parameters used for the RT-PCR reaction on the ABI Prism 7700 Sequence Detection System were optimized for HCV detection:



		Time
Cycle	Temperature (° C.)	(Minutes)	Repeat	Reaction

Hold	50	2		Initial Step
Hold	60	30		Reverse
				Transcription
Hold	95	5		UNG Deactivation
Cycle	95	0:15	2	Melt
	60	1		Anneal/Extend
Cycle	90	0:15	40	Melt
	60	1		Anneal/Extend

Quantification is based on the threshold cycle, where the amplification plot crosses a defined fluorescence threshold. Comparison of the threshold cycles provides a highly sensitive measure of relative template concentration in different samples. Monitoring during early cycles, when PCR fidelity is at its highest, provides precise data for accurate quantification. The relative template concentration can be converted to RNA copy numbers by employing a standard curve of HCV RNA with known copy number (FIG. 7). [0125]

Example 10

A specific HCV NS3 protease anti-viral compound inhibits replication of the HCV replicon in S22.3 cell lines. [0126]
In order to determine the effect of a specific HCV NS3 protease anti-viral compound on replicon levels in S22.3 cells, the cells were seeded in 24 Well Cell Culture Cluster at 5×10[0127] ⁴cells per well in 500 μL of DMEM complemented with 10% FBS, PenStrep and 1 μg/mL Geneticin. Cells were incubated until compound addition in a 5% CO₂incubator at 37° C. The dose-response curve of the inhibitor displayed 11 concentrations resulting from serial two-fold dilutions (1:1). The starting concentration of compound A was 100 nM. One control well (without any compound) was also included in the course of the experiment. The 24 well plates were incubated for 4 days in a 5% CO₂incubator at 37° C. Following a 4 day incubation period, the cells were washed once with PBS and RNA was extracted with the RNeasy® Mini Kit and Qiashredder® from Qiagen. RNA from each well was eluted in 50uL of H₂O. The RNA was quantified by optical density at 260 nm on a Cary 1 E UV-Visible Spectrophotometer. 50 ng of RNA from each well was used to quantify the HCV replicon RNA copy number as detailed in Example 6. The level of inhibition (% inhibition) of each well containing inhibitor was calculated with the following equation (CN═HCV Replicon copy number): $% \cdot inhibition = (\frac{CN \cdot control - CN \cdot well}{CN \cdot control}) * 100$
The calculated % inhibition values were then used to determine IC[0128] ₅₀, slope factor (n) and maximum inhibition (I_max) by the non-linear regression routine NLIN procedure of SAS using the following equation: $% \cdot inhibition = \frac{I_{\max} \times {[inhibitor]}^{n}}{{[inhibitor]}^{n} + {IC}_{50}^{n}}$
Compound A was tested in the assay at least 4 times. The IC[0129] ₅₀curves were analyzed individually by the SAS nonlinear regression analysis. FIG. 8 shows a typical curve and Table 2 shows the individual and average IC₅₀values of compound A. The average IC₅₀of compound A in the replication assay was 1.1 nM.

TABLE 2

IC₅₀of compound A in the S22.3 Cell line Replicon Assay.

Compound IC₅₀(nM) Average IC₅₀(nM)

A 1.2

1.2

1.0

0.9

1.1 ± 0.2

DISCUSSION

The reproducible and robust ex vivo propagation of hepatitis C virus, to levels required for the accurate testing of potential anti-viral compounds, has not been achieved with any system. As an alternative approach to studying the molecular mechanisms of hepatitis C virus RNA replication, selectable self-replicating bi-cistronic RNAs were developed (Lohman et al., 1999, Science 285, 110-113; Bartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844 CA 2,303,526). Minimally, these replicons encode for some or all of the non-structural proteins and also carry a selectable marker such as the neomycin phosphotransferase. Though intracellular steady-state levels of these sub-genomic replicon RNAs among the selected clones is moderate to high, the frequency of generating G418-resistant colonies upon transfection of the consensus RNA described by Lohman et al. or Bartenschlager, R. et al., 1993, J. Virol., 67, 3835-3844 is very low. Less than 100 colonies are generated when 8 million cells are transfected with 1 μg of in vitro transcribed bi-cistronic replicon RNA. A low efficiency of colony formation was first noted by Lohmann et al (1999 et al, Science 285,110-113). Since then, Lohmann et al. (2001) J. Virol. 1437-1449, Blight et al. 2000, Science 290, 1972-1974, and Guo et al., (2001) J. Virol. 8516-8523, have isolated sub-genomic RNAs with markedly improved efficiencies in the colony formation assay. Lohmann et al., 1999 Science 285,110-113 originally reported that selection of sub genomic replicons may not involve the selection of adaptive mutants as serially passaged RNA did not demonstrate an improved transfection efficiency. Nevertheless, in an effort to characterize the function and fitness of replicating HCV RNA, we serially passaged the replicon RNA that was isolated from the first selected cell-line. Notably, a significant increase in colony forming efficiency was obtained from this experiment, even though the quantity of replicon RNA was orders of magnitude lower than originally used to transfect the in vitro transcribed RNA. Furthermore, a second round serial passage of replicon RNA from this first generation clone into naive Huh-7 cells provided for yet another increase in colony formation efficiency (Table 1). [0130]
Our analysis of replicating HCV RNAs identified several adaptive mutations that enhance the efficiency of colony formation by up to 4 orders of magnitude. Adaptive mutations were found in many non-structural proteins, as well as in the 5′ non-translated region. The substitution of the 5′-GG doublet for a 5′-A as the inaugurating nucleotide of the [0131] HCV 5′-UTR is a variant of the HCV genome that has not been previously described, despite the sequencing of innumerable genotypes and subtypes from across the world. Our original replicon that carried a 5′-GG evolved to variants with either a single 5′-A or 5′-G, both of which showed equal transduction efficiency. We describe here the first report of a HCV genome that can tolerate and stably maintain a 5′A extremity. Moreover, we were successful in re-introducing this defined single nucleotide substitution into our cDNA clone and generate in vitro transcribed RNA harboring such an extremity to confirm that a 5′A functions as efficiently as a 5′G.
We have identified adaptive amino acid substitutions in the HCV non-structural proteins NS3, NS4A and NS5A in the R3 replicon, and a substitution in NS5B in the R7 clone (see FIG. 5B). These mutations, particularly the combination defined by the R3-rep (SEQ ID NO. 7), when reconstituted into a cDNA clone and transcribed onto a RNA replicon, result in a significantly enhanced transduction efficiency of up to 20,000 fold from the original wild type APGK12 replicon RNA. However, the steady state levels of intracellular replicon RNA were comparable from each of the different isolated clones. This result suggests that the increase in replication efficiency by the adaptive mutations does not result in higher stable intracellular RNA levels due to higher RNA replication, but rather confers increased permissivity for establishing the replicon in a greater number of Huh7 cells. Such a phenotype may be manifested transiently, through an initial increase of the amount of de novo replication, that is required to surpass a defined threshold to establish persistently replicating RNAs within a population of dividing cells. [0132]
Recently three other groups also identified other distinct adaptive mutants. Lohmann et al. (2000) reported enhanced transduction efficiencies of up to 10,000 fold with mutations in NS3, NS4B, NS5A and NS5B. Blight et al. 2000, Science 290:1972-1974 reported an augmentation of transduction efficiencies up to 20,000 fold with a single mutation in NS5A whereas Guo et al., (2001) J. Virol. 8516-8523 reported increases in transduction efficiencies of 5,000-10,000 fold with a deletion of a single amino acid in NS5A. The amino acid substitutions that we describe here have not previously been identified as adaptive mutants that enhance the efficiency of RNA transfection and/or replication. One exception is the mutation of E1202G in NS3 that we found in both the R7 and R16 replicons. This adaptation was previously described by Guo et al., (2001) J. Virol. 8516-8523 and Krieger et al (2001) J. Virol. 4614-4624. All other adaptive mutations, without exception, described herein are unpublished. [0133]
The development of selectable subgenomic HCV replicons has provided for potential avenues of exploration on HCV RNA replication, persistence, and pathogenesis in cultured cells. However, the low transduction efficiency with the HCV RNA-containing replicons as originally described (Lohmann et al., 1999 Science 285: 110-113) showed that it was not a practical system for reverse genetics studies. The adaptive mutants described herein overcome the low transduction efficiency. In light of the recent descriptions of adaptive mutants by other groups, we note that adaptation can be achieved by distinct mutations in different HCV NS proteins, although the level of adaptation can vary drastically. The replicons encoding adaptive mutants that are described herein are ideally suited for reverse genetic studies to identify novel HCV targets or host cell targets that may modulate HCV RNA replication or HCV replicon RNA colony formation. The adapted and highly efficient replicons are suitable tools for characterizing subtle genotypic or phenotypic changes that affect an easily quantifiable transduction efficiency. [0134]
Lastly, we have used our adapted HCV sub genomic replicon cell-line to demonstrate the proficient inhibition of HCV RNA replication by a specific small molecule inhibitor of the HCV NS3 protease. This is the first demonstration that an antiviral, designed to specifically inhibit one of the HCV non-structural proteins, inhibits HCV RNA replication in cell culture. Moreover, this compound and our S22.3 cell line validate the proposal that RNA replication is directed by the HCV non-structural proteins NS3 to NS5B. The assay that we have described and validated will be extremely useful in characterizing other inhibitors of HCV non-structural protein function in cell culture in a high throughput fashion. [0135]
All references found throughout the present disclosure are herein incorporated by reference whether they be found in the following list or not. [0136]
1 25 1 8639 DNA HCV CDS (1803)...(8408) 1 ggccagcccc cgattggggg cgacactcca ccatagatca ctcccctgtg aggaactact 60 gtcttcacgc agaaagcgtc tagccatggc gttagtatga gtgtcgtgca gcctccagga 120 ccccccctcc cgggagagcc atagtggtct gcggaaccgg tgagtacacc ggaattgcca 180 ggacgaccgg gtcctttctt ggatcaaccc gctcaatgcc tggagatttg ggcgtgcccc 240 cgcgagactg ctagccgagt agtgttgggt cgcgaaaggc cttgtggtac tgcctgatag 300 ggtgcttgcg agtgccccgg gaggtctcgt agaccgtgca ccatgagcac gaatcctaaa 360 cctcaaagaa aaaccaaagg gcgcgccatg attgaacaag atggattgca cgcaggttct 420 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc 480 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc 540 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc 600 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg 660 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag 720 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc 780 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt 840 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc 900 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc 960 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg 1020 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag 1080 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg 1140 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagttcgcgc ccagatgtta 1200 acagaccaca acggtttccc tctagcggga tcaattccgc ccccccccct aacgttactg 1260 gccgaagccg cttggaataa ggccggtgtg cgtttgtcta tatgttattt tccaccatat 1320 tgccgtcttt tggcaatgtg agggcccgga aacctggccc tgtcttcttg acgagcattc 1380 ctaggggtct ttcccctctc gccaaaggaa tgcaaggtct gttgaatgtc gtgaaggaag 1440 cagttcctct ggaagcttct tgaagacaaa caacgtctgt agcgaccctt tgcaggcagc 1500 ggaacccccc acctggcgac aggtgcctct gcggccaaaa gccacgtgta taagatacac 1560 ctgcaaaggc ggcacaaccc cagtgccacg ttgtgagttg gatagttgtg gaaagagtca 1620 aatggctctc ctcaagcgta ttcaacaagg ggctgaagga tgcccagaag gtaccccatt 1680 gtatgggatc tgatctgggg cctcggtgca catgctttac atgtgtttag tcgaggttaa 1740 aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa cacgataata 1800 cc atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta 1847 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val 1 5 10 15 ggt ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct 1895 Gly Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala 20 25 30 agg ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac 1943 Arg Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His 35 40 45 ttg caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc 1991 Leu Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala 50 55 60 gtc atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc 2039 Val Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile 65 70 75 acc aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct 2087 Thr Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala 80 85 90 95 ggt ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt 2135 Gly Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg 100 105 110 gca tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg 2183 Ala Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met 115 120 125 gct ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat 2231 Ala Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His 130 135 140 ctc acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg 2279 Leu Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala 145 150 155 gtg gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc 2327 Val Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile 160 165 170 175 acc tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg 2375 Thr Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu 180 185 190 ccc gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac 2423 Pro Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp 195 200 205 agc ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac 2471 Ser Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr 210 215 220 tcc caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctc aca 2519 Ser Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr 225 230 235 ggc cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc 2567 Gly Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr 240 245 250 255 gca aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act 2615 Ala Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr 260 265 270 gtc tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca 2663 Val Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro 275 280 285 atc acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa 2711 Ile Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln 290 295 300 gcg ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg 2759 Ala Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser 305 310 315 gac ctt tac ttg gtc acg agg cat gcc gat gtc att ccg gtg cgc cgg 2807 Asp Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg 320 325 330 335 cgg ggc gac agc agg ggg agc cta ctc tcc ccc agg ccc gtc tcc tac 2855 Arg Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr 340 345 350 ttg aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct 2903 Leu Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala 355 360 365 gtg ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg 2951 Val Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala 370 375 380 gtg gac ttt gta ccc gtc gag tct atg gaa acc act atg cgg tcc ccg 2999 Val Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro 385 390 395 gtc ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag 3047 Val Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln 400 405 410 415 gtg gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg 3095 Val Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val 420 425 430 ccg gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg 3143 Pro Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro 435 440 445 tcc gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat 3191 Ser Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His 450 455 460 ggt atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt 3239 Gly Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly 465 470 475 gcc ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt 3287 Ala Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly 480 485 490 495 tgc tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca 3335 Cys Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser 500 505 510 act gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg 3383 Thr Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala 515 520 525 gag acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg 3431 Glu Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro 530 535 540 gga tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc 3479 Gly Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser 545 550 555 agc act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc 3527 Ser Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr 560 565 570 575 atc aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt 3575 Ile Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys 580 585 590 gat gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca 3623 Asp Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala 595 600 605 tat tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc 3671 Tyr Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val 610 615 620 att gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc 3719 Ile Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe 625 630 635 gac tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc 3767 Asp Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe 640 645 650 655 agc ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac 3815 Ser Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp 660 665 670 gcg gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg 3863 Ala Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met 675 680 685 ggc att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc 3911 Gly Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe 690 695 700 gat tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac 3959 Asp Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr 705 710 715 gag ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac 4007 Glu Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn 720 725 730 735 aca cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc 4055 Thr Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser 740 745 750 gtc ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act 4103 Val Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr 755 760 765 aag cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg 4151 Lys Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr 770 775 780 gtg tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg 4199 Val Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp 785 790 795 aag tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg 4247 Lys Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu 800 805 810 815 ctg tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc 4295 Leu Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro 820 825 830 ata acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc 4343 Ile Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val 835 840 845 acg agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg 4391 Thr Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala 850 855 860 tat tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg 4439 Tyr Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu 865 870 875 tcc gga aag ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag 4487 Ser Gly Lys Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu 880 885 890 895 ttc gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag 4535 Phe Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln 900 905 910 gga atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg 4583 Gly Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu 915 920 925 caa aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc 4631 Gln Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser 930 935 940 aag tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc 4679 Lys Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe 945 950 955 atc agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac 4727 Ile Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn 960 965 970 975 ccc gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg 4775 Pro Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro 980 985 990 ctc acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg 4823 Leu Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val 995 1000 1005 gcc gcc caa ctt gct cct ccc agc gct gct tct gct ttc gta ggc gcc 4871 Ala Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala 1010 1015 1020 ggc atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt 4919 Gly Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu 1025 1030 1035 gtg gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg 4967 Val Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val 1040 1045 1050 1055 gcc ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt 5015 Ala Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val 1060 1065 1070 aac cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc 5063 Asn Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val 1075 1080 1085 gtg tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct 5111 Val Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala 1090 1095 1100 gtg cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac 5159 Val Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His 1105 1110 1115 gtc tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc 5207 Val Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val 1120 1125 1130 1135 act cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt 5255 Thr Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu 1140 1145 1150 cac cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg 5303 His Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp 1155 1160 1165 cta aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag 5351 Leu Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys 1170 1175 1180 acc tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc 5399 Thr Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe 1185 1190 1195 ttc tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc 5447 Phe Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile 1200 1205 1210 1215 atg caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa 5495 Met Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys 1220 1225 1230 aac ggt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg 5543 Asn Gly Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp 1235 1240 1245 cat gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc 5591 His Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro 1250 1255 1260 tcc ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag 5639 Ser Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu 1265 1270 1275 gag tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggc 5687 Glu Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly 1280 1285 1290 1295 atg acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa 5735 Met Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu 1300 1305 1310 ttc ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg 5783 Phe Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala 1315 1320 1325 tgc aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat 5831 Cys Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn 1330 1335 1340 caa tac ctg gtt ggg tca cag ctc cca tgc gag ccc gaa ccg gac gta 5879 Gln Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val 1345 1350 1355 gca gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag 5927 Ala Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu 1360 1365 1370 1375 acg gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc 5975 Thr Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser 1380 1385 1390 tca tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act 6023 Ser Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr 1395 1400 1405 acc cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg 6071 Thr Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu 1410 1415 1420 tgg cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat 6119 Trp Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn 1425 1430 1435 aag gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat 6167 Lys Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp 1440 1445 1450 1455 gag agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa 6215 Glu Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys 1460 1465 1470 ttc cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca 6263 Phe Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro 1475 1480 1485 ctg tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac 6311 Leu Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His 1490 1495 1500 ggg tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg 6359 Gly Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg 1505 1510 1515 agg aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg 6407 Arg Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu 1520 1525 1530 1535 gcg gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc 6455 Ala Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val 1540 1545 1550 gac agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc 6503 Asp Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly 1555 1560 1565 gac gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag 6551 Asp Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu 1570 1575 1580 ggg gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta 6599 Gly Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val 1585 1590 1595 agc gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca 6647 Ser Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr 1600 1605 1610 1615 tgg aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg 6695 Trp Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu 1620 1625 1630 ccc atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc 6743 Pro Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val 1635 1640 1645 tat gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc 6791 Tyr Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr 1650 1655 1660 ttt gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag 6839 Phe Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys 1665 1670 1675 gag atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg 6887 Glu Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val 1680 1685 1690 1695 gag gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt 6935 Glu Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe 1700 1705 1710 ggc tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac 6983 Gly Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn 1715 1720 1725 cac atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca 7031 His Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro 1730 1735 1740 att gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca 7079 Ile Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro 1745 1750 1755 gag aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg 7127 Glu Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu 1760 1765 1770 1775 ggg gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc 7175 Gly Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr 1780 1785 1790 ctc cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct 7223 Leu Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro 1795 1800 1805 gga cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc 7271 Gly Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys 1810 1815 1820 cct atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act 7319 Pro Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr 1825 1830 1835 gag aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg 7367 Glu Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu 1840 1845 1850 1855 gcc ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac 7415 Ala Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr 1860 1865 1870 atc ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc 7463 Ile Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg 1875 1880 1885 cgg tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc 7511 Arg Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu 1890 1895 1900 aca tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag 7559 Thr Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln 1905 1910 1915 gac tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa 7607 Asp Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu 1920 1925 1930 1935 agc gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag 7655 Ser Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu 1940 1945 1950 gct atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa 7703 Ala Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu 1955 1960 1965 tac gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg 7751 Tyr Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala 1970 1975 1980 cac gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc 7799 His Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr 1985 1990 1995 acc ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc 7847 Thr Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val 2000 2005 2010 2015 aat tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca 7895 Asn Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala 2020 2025 2030 agg atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa 7943 Arg Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu 2035 2040 2045 caa ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc 7991 Gln Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser 2050 2055 2060 att gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt 8039 Ile Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu 2065 2070 2075 agc gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg 8087 Ser Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val 2080 2085 2090 2095 gct tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga 8135 Ala Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg 2100 2105 2110 cat cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg 8183 His Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg 2115 2120 2125 gct gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag 8231 Ala Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys 2130 2135 2140 ctc aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc 8279 Leu Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser 2145 2150 2155 tgg ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct 8327 Trp Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser 2160 2165 2170 2175 cgt gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta 8375 Arg Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val 2180 2185 2190 ggg gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8428 Gly Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt 8488 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct 8548 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga 8608 gtgctgatac tggcctctct gcagatcaag t 8639 2 8642 DNA HCV CDS (1802)...(8407) variation 6268 r = a or g 2 accagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc agg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga arg ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Xaa Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag ccc gaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 ttr gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Xaa Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttcccct tttttttttt tttttttttc tttttttttt 8487 tttttttttt tttttttttc tccttttttt tcctcttttt ttccttttct ttcctttggt 8547 ggctccatct tagccctagt cacggctagc tgtgaaaggt ccgtgagccg cttgactgca 8607 gagagtgctg atactggcct ctctgcagat caagt 8642 3 2201 PRT HCV VARIANT 882 Xaa is Lys or Arg 3 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser Val 740 745 750 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 Gly Xaa Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 Xaa Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 Val Gly Ile Tyr Leu Leu Pro Asn Arg 2195 2200 4 8643 DNA HCV CDS (1802)...(8407) 4 accagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc gcaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaga 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg aag cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Lys His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc cgg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag ggc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Gly Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga agg ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Arg Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tcc gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag gcc 5353 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Ala 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cga gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag ccc gaa ctg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Leu Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 ttg gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 acg ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt ttttcccttt tttttttttt tttttttttt tttttttttt 8487 tttttttttt tttttttttt ttttcttttt tcccaatttt tttccttttc tttcctttgg 8547 tggctccatc ttagccctag tcacggctag ctgtgaaagg tccgtgagcc gcttgactgc 8607 agagagtgct gatactggcc tctctgcaga tcaagt 8643 5 8648 DNA HCV CDS (1802)...(8407) 5 gccagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc agg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cgg ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gga acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Gly Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aag ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Lys Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg gta tgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Val Cys Thr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag ccc gaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg ccc tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gct tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc acg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Thr Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccctt ttttcccttt tttttttttt tttttttttt 8487 tttttttttt tttttttttt ttccccccct tttttcccct ttttttttcc ttttctttcc 8547 tttggtggct ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg 8607 actgcagaga gtgctgatac tggcctctct gcagatcaag t 8648 6 8638 DNA HCV CDS (1802)...(8407) 6 accagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac ggc agg ggg agc cta ctc tcc ccc agg ccc gtc tcc tac ttg 2857 Gly Asp Gly Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gga acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Gly Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aag ccg gcc atc att ccc gac agg gaa gtc ttt tac cgg gag ttc 4489 Gly Lys Pro Ala Ile Ile Pro Asp Arg Glu Val Phe Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 cgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Arg Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag ccc gaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ctg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg cct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Pro Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttttttt tttttttttt tttttttttt tttttttttt 8487 tttttttttt tttttttttt ttttttcctc ttttttttcc ttttctttcc tttggtggct 8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga 8607 gtgctgatac tggcctctct gcagatcaag t 8638 7 8638 DNA HCV CDS (1802)...(8407) 7 gccagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc acc agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg aag cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Lys His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc cgg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgc gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag ggc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Gly Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gct gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga agg ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Arg Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgt gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tcc gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag gcc 5353 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Ala 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cga gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ccg gtt ggg tca cag ctc cca tgc gag ccc gaa ctg gac gta gca 5881 Tyr Pro Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Leu Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gag aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt 8487 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct 8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga 8607 gtgctgatac tggcctctct gcagatcaag t 8638 8 6 DNA HCV 8 accagc 6 9 63 DNA HCV 9 gaattccaga tggcgcgccc agatgttaac cagatccatg gcacactcta gagtactgtc 60 gac 63 10 33 DNA HCV 10 cggaatcgtt aacagaccac aacggtttcc ctc 33 11 30 DNA HCV 11 ggcgtaccca tggtattatc gtgtttttca 30 12 45 DNA HCV 12 gcatatgaat tctaatacga ctcactatag gccagccccc gattg 45 13 45 DNA HCV 13 ggcgcgccct ttggtttttc tttgaggttt aggattcgtg ctcat 45 14 36 DNA HCV 14 aaagggcgca tgattgaaca agatggattg cacgca 36 15 39 DNA HCV 15 gcatatgtta actcagaaga actcgtcaag aaggcgata 39 16 45 DNA HCV 16 gcatatgaat tctaatacga ctcactatag gccagccccc gattg 45 17 30 DNA HCV 17 acgcagaaag cgtctagcca tggcgttagt 30 18 30 DNA HCV 18 tcccggggca ctcgcaagca ccctatcagg 30 19 26 DNA HCV Label with FAM fluorescence reporter dye 19 tggtctgcgg aacgggtgag tacacc 26 20 45 DNA HCV 20 gtggacgaat tctaatacga ctcactataa ccagcccccg attgg 45 21 27 DNA HCV 21 ggaacgcccg tcgtggccag ccacgat 27 22 23 DNA HCV 22 gtcgtcttct ctgacatgga gac 23 23 27 DNA HCV 23 gagttgctca gtggattgat gggcagc 27 24 8638 DNA HCV CDS (1802)...(8407) 24 accagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc act agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg agg cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Arg His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc agg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgt gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag agc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Ser Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gcc gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga aag ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Lys Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgc gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tct gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag acc 5353 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Thr 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 ggt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Gly Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cgg gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ctg gtt ggg tca cag ctc cca tgc gag ccc gaa ccg gac gta gca 5881 Tyr Leu Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Pro Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gaa aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt 8487 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct 8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga 8607 gtgctgatac tggcctctct gcagatcaag t 8638 25 8638 DNA HCV CDS (1802)...(8407) 25 accagccccc gattgggggc gacactccac catagatcac tcccctgtga ggaactactg 60 tcttcacgca gaaagcgtct agccatggcg ttagtatgag tgtcgtgcag cctccaggac 120 cccccctccc gggagagcca tagtggtctg cggaaccggt gagtacaccg gaattgccag 180 gacgaccggg tcctttcttg gatcaacccg ctcaatgcct ggagatttgg gcgtgccccc 240 gcgagactgc tagccgagta gtgttgggtc gcgaaaggcc ttgtggtact gcctgatagg 300 gtgcttgcga gtgccccggg aggtctcgta gaccgtgcac catgagcacg aatcctaaac 360 ctcaaagaaa aaccaaaggg cgcgccatga ttgaacaaga tggattgcac gcaggttctc 420 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct 480 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg 540 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca 600 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc 660 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga 720 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc 780 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc 840 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg 900 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct 960 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc 1020 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc 1080 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc 1140 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agttcgcgcc cagatgttaa 1200 cagaccacaa cggtttccct ctagcgggat caattccgcc ccccccccta acgttactgg 1260 ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt ccaccatatt 1320 gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga cgagcattcc 1380 taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg tgaaggaagc 1440 agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt gcaggcagcg 1500 gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat aagatacacc 1560 tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg aaagagtcaa 1620 atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg taccccattg 1680 tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt cgaggttaaa 1740 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgataatac 1800 c atg gac cgg gag atg gca gca tcg tgc gga ggc gcg gtt ttc gta ggt 1849 Met Asp Arg Glu Met Ala Ala Ser Cys Gly Gly Ala Val Phe Val Gly 1 5 10 15 ctg ata ctc ttg acc ttg tca ccg cac tat aag ctg ttc ctc gct agg 1897 Leu Ile Leu Leu Thr Leu Ser Pro His Tyr Lys Leu Phe Leu Ala Arg 20 25 30 ctc ata tgg tgg tta caa tat ttt atc acc agg gcc gag gca cac ttg 1945 Leu Ile Trp Trp Leu Gln Tyr Phe Ile Thr Arg Ala Glu Ala His Leu 35 40 45 caa gtg tgg atc ccc ccc ctc aac gtt cgg ggg ggc cgc gat gcc gtc 1993 Gln Val Trp Ile Pro Pro Leu Asn Val Arg Gly Gly Arg Asp Ala Val 50 55 60 atc ctc ctc acg tgc gcg atc cac cca gag cta atc ttt acc atc acc 2041 Ile Leu Leu Thr Cys Ala Ile His Pro Glu Leu Ile Phe Thr Ile Thr 65 70 75 80 aaa atc ttg ctc gcc ata ctc ggt cca ctc atg gtg ctc cag gct ggt 2089 Lys Ile Leu Leu Ala Ile Leu Gly Pro Leu Met Val Leu Gln Ala Gly 85 90 95 ata acc aaa gtg ccg tac ttc gtg cgc gca cac ggg ctc att cgt gca 2137 Ile Thr Lys Val Pro Tyr Phe Val Arg Ala His Gly Leu Ile Arg Ala 100 105 110 tgc atg ctg gtg cgg aag gtt gct ggg ggt cat tat gtc caa atg gct 2185 Cys Met Leu Val Arg Lys Val Ala Gly Gly His Tyr Val Gln Met Ala 115 120 125 ctc atg aag ttg gcc gca ctg aca ggt acg tac gtt tat gac cat ctc 2233 Leu Met Lys Leu Ala Ala Leu Thr Gly Thr Tyr Val Tyr Asp His Leu 130 135 140 acc cca ctg cgg gac tgg gcc cac gcg ggc cta cga gac ctt gcg gtg 2281 Thr Pro Leu Arg Asp Trp Ala His Ala Gly Leu Arg Asp Leu Ala Val 145 150 155 160 gca gtt gag ccc gtc gtc ttc tct gat atg gag acc aag gtt atc acc 2329 Ala Val Glu Pro Val Val Phe Ser Asp Met Glu Thr Lys Val Ile Thr 165 170 175 tgg ggg gca gac acc gcg gcg tgt ggg gac atc atc ttg ggc ctg ccc 2377 Trp Gly Ala Asp Thr Ala Ala Cys Gly Asp Ile Ile Leu Gly Leu Pro 180 185 190 gtc tcc gcc cgc agg ggg agg gag ata cat ctg gga ccg gca gac agc 2425 Val Ser Ala Arg Arg Gly Arg Glu Ile His Leu Gly Pro Ala Asp Ser 195 200 205 ctt gaa ggg cag ggg tgg cga ctc ctc gcg cct att acg gcc tac tcc 2473 Leu Glu Gly Gln Gly Trp Arg Leu Leu Ala Pro Ile Thr Ala Tyr Ser 210 215 220 caa cag acg cga ggc cta ctt ggc tgc atc atc acc agc ctc aca ggc 2521 Gln Gln Thr Arg Gly Leu Leu Gly Cys Ile Ile Thr Ser Leu Thr Gly 225 230 235 240 cgg gac agg aac cag gtc gag ggg gag gtc caa gtg gtc tcc acc gca 2569 Arg Asp Arg Asn Gln Val Glu Gly Glu Val Gln Val Val Ser Thr Ala 245 250 255 aca caa tct ttc ctg gcg acc tgc gtc aat ggc gtg tgt tgg act gtc 2617 Thr Gln Ser Phe Leu Ala Thr Cys Val Asn Gly Val Cys Trp Thr Val 260 265 270 tat cat ggt gcc ggc tca aag acc ctt gcc ggc cca aag ggc cca atc 2665 Tyr His Gly Ala Gly Ser Lys Thr Leu Ala Gly Pro Lys Gly Pro Ile 275 280 285 acc caa atg tac acc aat gtg gac cag gac ctc gtc ggc tgg caa gcg 2713 Thr Gln Met Tyr Thr Asn Val Asp Gln Asp Leu Val Gly Trp Gln Ala 290 295 300 ccc ccc ggg gcg cgt tcc ttg aca cca tgc acc tgc ggc agc tcg gac 2761 Pro Pro Gly Ala Arg Ser Leu Thr Pro Cys Thr Cys Gly Ser Ser Asp 305 310 315 320 ctt tac ttg gtc acg aag cat gcc gat gtc att ccg gtg cgc cgg cgg 2809 Leu Tyr Leu Val Thr Lys His Ala Asp Val Ile Pro Val Arg Arg Arg 325 330 335 ggc gac agc agg ggg agc cta ctc tcc ccc cgg ccc gtc tcc tac ttg 2857 Gly Asp Ser Arg Gly Ser Leu Leu Ser Pro Arg Pro Val Ser Tyr Leu 340 345 350 aag ggc tct tcg ggc ggt cca ctg ctc tgc ccc tcg ggg cac gct gtg 2905 Lys Gly Ser Ser Gly Gly Pro Leu Leu Cys Pro Ser Gly His Ala Val 355 360 365 ggc atc ttt cgg gct gcc gtg tgc acc cga ggg gtt gcg aag gcg gtg 2953 Gly Ile Phe Arg Ala Ala Val Cys Thr Arg Gly Val Ala Lys Ala Val 370 375 380 gac ttt gta ccc gtc gag tct atg gaa acc act atg cgg tcc ccg gtc 3001 Asp Phe Val Pro Val Glu Ser Met Glu Thr Thr Met Arg Ser Pro Val 385 390 395 400 ttc acg gac aac tcg tcc cct ccg gcc gta ccg cag aca ttc cag gtg 3049 Phe Thr Asp Asn Ser Ser Pro Pro Ala Val Pro Gln Thr Phe Gln Val 405 410 415 gcc cat cta cac gcc cct act ggt agc ggc aag agc act aag gtg ccg 3097 Ala His Leu His Ala Pro Thr Gly Ser Gly Lys Ser Thr Lys Val Pro 420 425 430 gct gcg tat gca gcc caa ggg tat aag gtg ctt gtc ctg aac ccg tcc 3145 Ala Ala Tyr Ala Ala Gln Gly Tyr Lys Val Leu Val Leu Asn Pro Ser 435 440 445 gtc gcc gcc acc cta ggt ttc ggg gcg tat atg tct aag gca cat ggt 3193 Val Ala Ala Thr Leu Gly Phe Gly Ala Tyr Met Ser Lys Ala His Gly 450 455 460 atc gac cct aac atc aga acc ggg gta agg acc atc acc acg ggt gcc 3241 Ile Asp Pro Asn Ile Arg Thr Gly Val Arg Thr Ile Thr Thr Gly Ala 465 470 475 480 ccc atc acg tac tcc acc tat ggc aag ttt ctt gcc gac ggt ggt tgc 3289 Pro Ile Thr Tyr Ser Thr Tyr Gly Lys Phe Leu Ala Asp Gly Gly Cys 485 490 495 tct ggg ggc gcc tat gac atc ata ata tgt gat gag tgc cac tca act 3337 Ser Gly Gly Ala Tyr Asp Ile Ile Ile Cys Asp Glu Cys His Ser Thr 500 505 510 gac tcg acc act atc ctg ggc atc ggc aca gtc ctg gac caa gcg gag 3385 Asp Ser Thr Thr Ile Leu Gly Ile Gly Thr Val Leu Asp Gln Ala Glu 515 520 525 acg gct gga gcg cga ctc gtc gtg ctc gcc acc gct acg cct ccg gga 3433 Thr Ala Gly Ala Arg Leu Val Val Leu Ala Thr Ala Thr Pro Pro Gly 530 535 540 tcg gtc acc gtg cca cat cca aac atc gag gag gtg gct ctg tcc agc 3481 Ser Val Thr Val Pro His Pro Asn Ile Glu Glu Val Ala Leu Ser Ser 545 550 555 560 act gga gaa atc ccc ttt tat ggc aaa gcc atc ccc atc gag acc atc 3529 Thr Gly Glu Ile Pro Phe Tyr Gly Lys Ala Ile Pro Ile Glu Thr Ile 565 570 575 aag ggg ggg agg cac ctc att ttc tgc cat tcc aag aag aaa tgc gat 3577 Lys Gly Gly Arg His Leu Ile Phe Cys His Ser Lys Lys Lys Cys Asp 580 585 590 gag ctc gcc gcg aag ctg tcc ggc ctc gga ctc aat gct gta gca tat 3625 Glu Leu Ala Ala Lys Leu Ser Gly Leu Gly Leu Asn Ala Val Ala Tyr 595 600 605 tac cgg ggc ctt gat gta tcc gtc ata cca act agc gga gac gtc att 3673 Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Ile 610 615 620 gtc gta gca acg gac gct cta atg acg ggc ttt acc ggc gat ttc gac 3721 Val Val Ala Thr Asp Ala Leu Met Thr Gly Phe Thr Gly Asp Phe Asp 625 630 635 640 tca gtg atc gac tgc aat aca tgt gtc acc cag aca gtc gac ttc agc 3769 Ser Val Ile Asp Cys Asn Thr Cys Val Thr Gln Thr Val Asp Phe Ser 645 650 655 ctg gac ccg acc ttc acc att gag acg acg acc gtg cca caa gac gcg 3817 Leu Asp Pro Thr Phe Thr Ile Glu Thr Thr Thr Val Pro Gln Asp Ala 660 665 670 gtg tca cgc tcg cag cgg cga ggc agg act ggt agg ggc agg atg ggc 3865 Val Ser Arg Ser Gln Arg Arg Gly Arg Thr Gly Arg Gly Arg Met Gly 675 680 685 att tac agg ttt gtg act cca gga gaa cgg ccc tcg ggc atg ttc gat 3913 Ile Tyr Arg Phe Val Thr Pro Gly Glu Arg Pro Ser Gly Met Phe Asp 690 695 700 tcc tcg gtt ctg tgc gag tgc tat gac gcg ggc tgt gct tgg tac gag 3961 Ser Ser Val Leu Cys Glu Cys Tyr Asp Ala Gly Cys Ala Trp Tyr Glu 705 710 715 720 ctc acg ccc gcc gag acc tca gtt agg ttg cgg gct tac cta aac aca 4009 Leu Thr Pro Ala Glu Thr Ser Val Arg Leu Arg Ala Tyr Leu Asn Thr 725 730 735 cca ggg ttg ccc gtc tgc cag gac cat ctg gag ttc tgg gag ggc gtc 4057 Pro Gly Leu Pro Val Cys Gln Asp His Leu Glu Phe Trp Glu Gly Val 740 745 750 ttt aca ggc ctc acc cac ata gac gcc cat ttc ttg tcc cag act aag 4105 Phe Thr Gly Leu Thr His Ile Asp Ala His Phe Leu Ser Gln Thr Lys 755 760 765 cag gca gga gac aac ttc ccc tac ctg gta gca tac cag gct acg gtg 4153 Gln Ala Gly Asp Asn Phe Pro Tyr Leu Val Ala Tyr Gln Ala Thr Val 770 775 780 tgc gcc agg gct cag gct cca cct cca tcg tgg gac caa atg tgg aag 4201 Cys Ala Arg Ala Gln Ala Pro Pro Pro Ser Trp Asp Gln Met Trp Lys 785 790 795 800 tgt ctc ata cgg cta aag cct acg ctg cac ggg cca acg ccc ctg ctg 4249 Cys Leu Ile Arg Leu Lys Pro Thr Leu His Gly Pro Thr Pro Leu Leu 805 810 815 tat agg ctg gga gcc gtt caa aac gag gtt act acc aca cac ccc ata 4297 Tyr Arg Leu Gly Ala Val Gln Asn Glu Val Thr Thr Thr His Pro Ile 820 825 830 acc aaa tac atc atg gca tgc atg tcg gct gac ctg gag gtc gtc acg 4345 Thr Lys Tyr Ile Met Ala Cys Met Ser Ala Asp Leu Glu Val Val Thr 835 840 845 agc acc tgg gtg ctg gta ggc gga gtc cta gca gct ctg gct gcg tat 4393 Ser Thr Trp Val Leu Val Gly Gly Val Leu Ala Ala Leu Ala Ala Tyr 850 855 860 tgc ctg aca aca ggc agc gtg gtc att gtg ggc agg atc atc ttg tcc 4441 Cys Leu Thr Thr Gly Ser Val Val Ile Val Gly Arg Ile Ile Leu Ser 865 870 875 880 gga agg ccg gcc atc att ccc gac agg gaa gtc ctt tac cgg gag ttc 4489 Gly Arg Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu Phe 885 890 895 gat gag atg gaa gag tgt gcc tca cac ctc cct tac atc gaa cag gga 4537 Asp Glu Met Glu Glu Cys Ala Ser His Leu Pro Tyr Ile Glu Gln Gly 900 905 910 atg cag ctc gcc gaa caa ttc aaa cag aag gca atc ggg ttg ctg caa 4585 Met Gln Leu Ala Glu Gln Phe Lys Gln Lys Ala Ile Gly Leu Leu Gln 915 920 925 aca gcc acc aag caa gcg gag gct gct gct ccc gtg gtg gaa tcc aag 4633 Thr Ala Thr Lys Gln Ala Glu Ala Ala Ala Pro Val Val Glu Ser Lys 930 935 940 tgg cgg acc ctc gaa gcc ttc tgg gcg aag cat atg tgg aat ttc atc 4681 Trp Arg Thr Leu Glu Ala Phe Trp Ala Lys His Met Trp Asn Phe Ile 945 950 955 960 agc ggg ata caa tat tta gca ggc ttg tcc act ctg cct ggc aac ccc 4729 Ser Gly Ile Gln Tyr Leu Ala Gly Leu Ser Thr Leu Pro Gly Asn Pro 965 970 975 gcg ata gca tca ctg atg gca ttc aca gcc tct atc acc agc ccg ctc 4777 Ala Ile Ala Ser Leu Met Ala Phe Thr Ala Ser Ile Thr Ser Pro Leu 980 985 990 acc acc caa cat acc ctc ctg ttt aac atc ctg ggg gga tgg gtg gcc 4825 Thr Thr Gln His Thr Leu Leu Phe Asn Ile Leu Gly Gly Trp Val Ala 995 1000 1005 gcc caa ctt gct cct ccc agc gct gct tcc gct ttc gta ggc gcc ggc 4873 Ala Gln Leu Ala Pro Pro Ser Ala Ala Ser Ala Phe Val Gly Ala Gly 1010 1015 1020 atc gct gga gcg gct gtt ggc agc ata ggc ctt ggg aag gtg ctt gtg 4921 Ile Ala Gly Ala Ala Val Gly Ser Ile Gly Leu Gly Lys Val Leu Val 1025 1030 1035 1040 gat att ttg gca ggt tat gga gca ggg gtg gca ggc gcg ctc gtg gcc 4969 Asp Ile Leu Ala Gly Tyr Gly Ala Gly Val Ala Gly Ala Leu Val Ala 1045 1050 1055 ttt aag gtc atg agc ggc gag atg ccc tcc acc gag gac ctg gtt aac 5017 Phe Lys Val Met Ser Gly Glu Met Pro Ser Thr Glu Asp Leu Val Asn 1060 1065 1070 cta ctc cct gct atc ctc tcc cct ggc gcc cta gtc gtc ggg gtc gtg 5065 Leu Leu Pro Ala Ile Leu Ser Pro Gly Ala Leu Val Val Gly Val Val 1075 1080 1085 tgc gca gcg ata ctg cgt cgg cac gtg ggc cca ggg gag ggg gct gtg 5113 Cys Ala Ala Ile Leu Arg Arg His Val Gly Pro Gly Glu Gly Ala Val 1090 1095 1100 cag tgg atg aac cgg ctg ata gcg ttc gct tcg cgg ggt aac cac gtc 5161 Gln Trp Met Asn Arg Leu Ile Ala Phe Ala Ser Arg Gly Asn His Val 1105 1110 1115 1120 tcc ccc acg cac tat gtg cct gag agc gac gct gca gca cgt gtc act 5209 Ser Pro Thr His Tyr Val Pro Glu Ser Asp Ala Ala Ala Arg Val Thr 1125 1130 1135 cag atc ctc tct agt ctt acc atc act cag ctg ctg aag agg ctt cac 5257 Gln Ile Leu Ser Ser Leu Thr Ile Thr Gln Leu Leu Lys Arg Leu His 1140 1145 1150 cag tgg atc aac gag gac tgc tcc acg cca tgc tcc ggc tcg tgg cta 5305 Gln Trp Ile Asn Glu Asp Cys Ser Thr Pro Cys Ser Gly Ser Trp Leu 1155 1160 1165 aga gat gtt tgg gat tgg ata tgc acg gtg ttg act gat ttc aag gcc 5353 Arg Asp Val Trp Asp Trp Ile Cys Thr Val Leu Thr Asp Phe Lys Ala 1170 1175 1180 tgg ctc cag tcc aag ctc ctg ccg cga ttg ccg gga gtc ccc ttc ttc 5401 Trp Leu Gln Ser Lys Leu Leu Pro Arg Leu Pro Gly Val Pro Phe Phe 1185 1190 1195 1200 tca tgt caa cgt ggg tac aag gga gtc tgg cgg ggc gac ggc atc atg 5449 Ser Cys Gln Arg Gly Tyr Lys Gly Val Trp Arg Gly Asp Gly Ile Met 1205 1210 1215 caa acc acc tgc cca tgt gga gca cag atc acc gga cat gtg aaa aac 5497 Gln Thr Thr Cys Pro Cys Gly Ala Gln Ile Thr Gly His Val Lys Asn 1220 1225 1230 tgt tcc atg agg atc gtg ggg cct agg acc tgt agt aac acg tgg cat 5545 Cys Ser Met Arg Ile Val Gly Pro Arg Thr Cys Ser Asn Thr Trp His 1235 1240 1245 gga aca ttc ccc att aac gcg tac acc acg ggc ccc tgc acg ccc tcc 5593 Gly Thr Phe Pro Ile Asn Ala Tyr Thr Thr Gly Pro Cys Thr Pro Ser 1250 1255 1260 ccg gcg cca aat tat tct agg gcg ctg tgg cgg gtg gct gct gag gag 5641 Pro Ala Pro Asn Tyr Ser Arg Ala Leu Trp Arg Val Ala Ala Glu Glu 1265 1270 1275 1280 tac gtg gag gtt acg cga gtg ggg gat ttc cac tac gtg acg ggc atg 5689 Tyr Val Glu Val Thr Arg Val Gly Asp Phe His Tyr Val Thr Gly Met 1285 1290 1295 acc act gac aac gta aag tgc ccg tgt cag gtt ccg gcc ccc gaa ttc 5737 Thr Thr Asp Asn Val Lys Cys Pro Cys Gln Val Pro Ala Pro Glu Phe 1300 1305 1310 ttc aca gaa gtg gat ggg gtg cgg ttg cac agg tac gct cca gcg tgc 5785 Phe Thr Glu Val Asp Gly Val Arg Leu His Arg Tyr Ala Pro Ala Cys 1315 1320 1325 aaa ccc ctc cta cgg gag gag gtc aca ttc ctg gtc ggg ctc aat caa 5833 Lys Pro Leu Leu Arg Glu Glu Val Thr Phe Leu Val Gly Leu Asn Gln 1330 1335 1340 tac ccg gtt ggg tca cag ctc cca tgc gag ccc gaa ctg gac gta gca 5881 Tyr Pro Val Gly Ser Gln Leu Pro Cys Glu Pro Glu Leu Asp Val Ala 1345 1350 1355 1360 gtg ctc act tcc atg ctc acc gac ccc tcc cac att acg gcg gag acg 5929 Val Leu Thr Ser Met Leu Thr Asp Pro Ser His Ile Thr Ala Glu Thr 1365 1370 1375 gct aag cgt agg ctg gcc agg gga tct ccc ccc tcc ttg gcc agc tca 5977 Ala Lys Arg Arg Leu Ala Arg Gly Ser Pro Pro Ser Leu Ala Ser Ser 1380 1385 1390 tca gct agc cag ctg tct gcg cct tcc ttg aag gca aca tgc act acc 6025 Ser Ala Ser Gln Leu Ser Ala Pro Ser Leu Lys Ala Thr Cys Thr Thr 1395 1400 1405 cgt cat gac tcc ccg gac gct gac ctc atc gag gcc aac ctc ctg tgg 6073 Arg His Asp Ser Pro Asp Ala Asp Leu Ile Glu Ala Asn Leu Leu Trp 1410 1415 1420 cgg cag gag atg ggc ggg aac atc acc cgc gtg gag tca gag aat aag 6121 Arg Gln Glu Met Gly Gly Asn Ile Thr Arg Val Glu Ser Glu Asn Lys 1425 1430 1435 1440 gta gta att ttg gac tct ttc gag ccg ctc caa gcg gag gag gat gag 6169 Val Val Ile Leu Asp Ser Phe Glu Pro Leu Gln Ala Glu Glu Asp Glu 1445 1450 1455 agg gaa gta tcc gtt ccg gcg gag atc ctg cgg agg tcc agg aaa ttc 6217 Arg Glu Val Ser Val Pro Ala Glu Ile Leu Arg Arg Ser Arg Lys Phe 1460 1465 1470 cct cga gcg atg ccc ata tgg gca cgc ccg gat tac aac cct cca ctg 6265 Pro Arg Ala Met Pro Ile Trp Ala Arg Pro Asp Tyr Asn Pro Pro Leu 1475 1480 1485 tta gag tcc tgg aag gac ccg gac tac gtc cct cca gtg gta cac ggg 6313 Leu Glu Ser Trp Lys Asp Pro Asp Tyr Val Pro Pro Val Val His Gly 1490 1495 1500 tgt cca ttg ccg cct gcc aag gcc cct ccg ata cca cct cca cgg agg 6361 Cys Pro Leu Pro Pro Ala Lys Ala Pro Pro Ile Pro Pro Pro Arg Arg 1505 1510 1515 1520 aag agg acg gtt gtc ctg tca gaa tct acc gtg tct tct gcc ttg gcg 6409 Lys Arg Thr Val Val Leu Ser Glu Ser Thr Val Ser Ser Ala Leu Ala 1525 1530 1535 gag ctc gcc aca aag acc ttc ggc agc tcc gaa tcg tcg gcc gtc gac 6457 Glu Leu Ala Thr Lys Thr Phe Gly Ser Ser Glu Ser Ser Ala Val Asp 1540 1545 1550 agc ggc acg gca acg gcc tct cct gac cag ccc tcc gac gac ggc gac 6505 Ser Gly Thr Ala Thr Ala Ser Pro Asp Gln Pro Ser Asp Asp Gly Asp 1555 1560 1565 gcg gga tcc gac gtt gag tcg tac tcc tcc atg ccc ccc ctt gag ggg 6553 Ala Gly Ser Asp Val Glu Ser Tyr Ser Ser Met Pro Pro Leu Glu Gly 1570 1575 1580 gag ccg ggg gat ccc gat ctc agc gac ggg tct tgg tct acc gta agc 6601 Glu Pro Gly Asp Pro Asp Leu Ser Asp Gly Ser Trp Ser Thr Val Ser 1585 1590 1595 1600 gag gag gct agt gag gac gtc gtc tgc tgc tcg atg tcc tac aca tgg 6649 Glu Glu Ala Ser Glu Asp Val Val Cys Cys Ser Met Ser Tyr Thr Trp 1605 1610 1615 aca ggc gcc ctg atc acg cca tgc gct gcg gag gaa acc aag ctg ccc 6697 Thr Gly Ala Leu Ile Thr Pro Cys Ala Ala Glu Glu Thr Lys Leu Pro 1620 1625 1630 atc aat gca ctg agc aac tct ttg ctc cgt cac cac aac ttg gtc tat 6745 Ile Asn Ala Leu Ser Asn Ser Leu Leu Arg His His Asn Leu Val Tyr 1635 1640 1645 gct aca aca tct cgc agc gca agc ctg cgg cag aag aag gtc acc ttt 6793 Ala Thr Thr Ser Arg Ser Ala Ser Leu Arg Gln Lys Lys Val Thr Phe 1650 1655 1660 gac aga ctg cag gtc ctg gac gac cac tac cgg gac gtg ctc aag gag 6841 Asp Arg Leu Gln Val Leu Asp Asp His Tyr Arg Asp Val Leu Lys Glu 1665 1670 1675 1680 atg aag gcg aag gcg tcc aca gtt aag gct aaa ctt cta tcc gtg gag 6889 Met Lys Ala Lys Ala Ser Thr Val Lys Ala Lys Leu Leu Ser Val Glu 1685 1690 1695 gaa gcc tgt aag ctg acg ccc cca cat tcg gcc aga tct aaa ttt ggc 6937 Glu Ala Cys Lys Leu Thr Pro Pro His Ser Ala Arg Ser Lys Phe Gly 1700 1705 1710 tat ggg gca aag gac gtc cgg aac cta tcc agc aag gcc gtt aac cac 6985 Tyr Gly Ala Lys Asp Val Arg Asn Leu Ser Ser Lys Ala Val Asn His 1715 1720 1725 atc cgc tcc gtg tgg aag gac ttg ctg gaa gac act gag aca cca att 7033 Ile Arg Ser Val Trp Lys Asp Leu Leu Glu Asp Thr Glu Thr Pro Ile 1730 1735 1740 gac acc acc atc atg gca aaa aat gag gtt ttc tgc gtc caa cca gag 7081 Asp Thr Thr Ile Met Ala Lys Asn Glu Val Phe Cys Val Gln Pro Glu 1745 1750 1755 1760 aag ggg ggc cgc aag cca gct cgc ctt atc gta ttc cca gat ttg ggg 7129 Lys Gly Gly Arg Lys Pro Ala Arg Leu Ile Val Phe Pro Asp Leu Gly 1765 1770 1775 gtt cgt gtg tgc gag aaa atg gcc ctt tac gat gtg gtc tcc acc ctc 7177 Val Arg Val Cys Glu Lys Met Ala Leu Tyr Asp Val Val Ser Thr Leu 1780 1785 1790 cct cag gcc gtg atg ggc tct tca tac gga ttc caa tac tct cct gga 7225 Pro Gln Ala Val Met Gly Ser Ser Tyr Gly Phe Gln Tyr Ser Pro Gly 1795 1800 1805 cag cgg gtc gag ttc ctg gtg aat gcc tgg aaa gcg aag aaa tgc cct 7273 Gln Arg Val Glu Phe Leu Val Asn Ala Trp Lys Ala Lys Lys Cys Pro 1810 1815 1820 atg ggc ttc gca tat gac acc cgc tgt ttt gac tca acg gtc act gag 7321 Met Gly Phe Ala Tyr Asp Thr Arg Cys Phe Asp Ser Thr Val Thr Glu 1825 1830 1835 1840 aat gac atc cgt gtt gag gag tca atc tac caa tgt tgt gac ttg gcc 7369 Asn Asp Ile Arg Val Glu Glu Ser Ile Tyr Gln Cys Cys Asp Leu Ala 1845 1850 1855 ccc gaa gcc aga cag gcc ata agg tcg ctc aca gag cgg ctt tac atc 7417 Pro Glu Ala Arg Gln Ala Ile Arg Ser Leu Thr Glu Arg Leu Tyr Ile 1860 1865 1870 ggg ggc ccc ctg act aat tct aaa ggg cag aac tgc ggc tat cgc cgg 7465 Gly Gly Pro Leu Thr Asn Ser Lys Gly Gln Asn Cys Gly Tyr Arg Arg 1875 1880 1885 tgc cgc gcg agc ggt gta ctg acg acc agc tgc ggt aat acc ctc aca 7513 Cys Arg Ala Ser Gly Val Leu Thr Thr Ser Cys Gly Asn Thr Leu Thr 1890 1895 1900 tgt tac ttg aag gcc gct gcg gcc tgt cga gct gcg aag ctc cag gac 7561 Cys Tyr Leu Lys Ala Ala Ala Ala Cys Arg Ala Ala Lys Leu Gln Asp 1905 1910 1915 1920 tgc acg atg ctc gta tgc gga gac gac ctt gtc gtt atc tgt gaa agc 7609 Cys Thr Met Leu Val Cys Gly Asp Asp Leu Val Val Ile Cys Glu Ser 1925 1930 1935 gcg ggg acc caa gag gac gag gcg agc cta cgg gcc ttc acg gag gct 7657 Ala Gly Thr Gln Glu Asp Glu Ala Ser Leu Arg Ala Phe Thr Glu Ala 1940 1945 1950 atg act aga tac tct gcc ccc cct ggg gac ccg ccc aaa cca gaa tac 7705 Met Thr Arg Tyr Ser Ala Pro Pro Gly Asp Pro Pro Lys Pro Glu Tyr 1955 1960 1965 gac ttg gag ttg ata aca tca tgc tcc tcc aat gtg tca gtc gcg cac 7753 Asp Leu Glu Leu Ile Thr Ser Cys Ser Ser Asn Val Ser Val Ala His 1970 1975 1980 gat gca tct ggc aaa agg gtg tac tat ctc acc cgt gac ccc acc acc 7801 Asp Ala Ser Gly Lys Arg Val Tyr Tyr Leu Thr Arg Asp Pro Thr Thr 1985 1990 1995 2000 ccc ctt gcg cgg gct gcg tgg gag aca gct aga cac act cca gtc aat 7849 Pro Leu Ala Arg Ala Ala Trp Glu Thr Ala Arg His Thr Pro Val Asn 2005 2010 2015 tcc tgg cta ggc aac atc atc atg tat gcg ccc acc ttg tgg gca agg 7897 Ser Trp Leu Gly Asn Ile Ile Met Tyr Ala Pro Thr Leu Trp Ala Arg 2020 2025 2030 atg atc ctg atg act cat ttc ttc tcc atc ctt cta gct cag gaa caa 7945 Met Ile Leu Met Thr His Phe Phe Ser Ile Leu Leu Ala Gln Glu Gln 2035 2040 2045 ctt gaa aaa gcc cta gat tgt cag atc tac ggg gcc tgt tac tcc att 7993 Leu Glu Lys Ala Leu Asp Cys Gln Ile Tyr Gly Ala Cys Tyr Ser Ile 2050 2055 2060 gag cca ctt gac cta cct cag atc att caa cga ctc cac ggc ctt agc 8041 Glu Pro Leu Asp Leu Pro Gln Ile Ile Gln Arg Leu His Gly Leu Ser 2065 2070 2075 2080 gca ttt tca ctc cat agt tac tct cca ggt gag atc aat agg gtg gct 8089 Ala Phe Ser Leu His Ser Tyr Ser Pro Gly Glu Ile Asn Arg Val Ala 2085 2090 2095 tca tgc ctc agg aaa ctt ggg gta ccg ccc ttg cga gtc tgg aga cat 8137 Ser Cys Leu Arg Lys Leu Gly Val Pro Pro Leu Arg Val Trp Arg His 2100 2105 2110 cgg gcc aga agt gtc cgc gct agg cta ctg tcc cag ggg ggg agg gct 8185 Arg Ala Arg Ser Val Arg Ala Arg Leu Leu Ser Gln Gly Gly Arg Ala 2115 2120 2125 gcc act tgt ggc aag tac ctc ttc aac tgg gca gta agg acc aag ctc 8233 Ala Thr Cys Gly Lys Tyr Leu Phe Asn Trp Ala Val Arg Thr Lys Leu 2130 2135 2140 aaa ctc act cca atc ccg gct gcg tcc cag ttg gat tta tcc agc tgg 8281 Lys Leu Thr Pro Ile Pro Ala Ala Ser Gln Leu Asp Leu Ser Ser Trp 2145 2150 2155 2160 ttc gtt gct ggt tac agc ggg gga gac ata tat cac agc ctg tct cgt 8329 Phe Val Ala Gly Tyr Ser Gly Gly Asp Ile Tyr His Ser Leu Ser Arg 2165 2170 2175 gcc cga ccc cgc tgg ttc atg tgg tgc cta ctc cta ctt tct gta ggg 8377 Ala Arg Pro Arg Trp Phe Met Trp Cys Leu Leu Leu Leu Ser Val Gly 2180 2185 2190 gta ggc atc tat cta ctc ccc aac cga tga acggggagct aaacactcca 8427 Val Gly Ile Tyr Leu Leu Pro Asn Arg * 2195 2200 ggccaatagg ccatcctgtt tttttccctt tttttttttc tttttttttt tttttttttt 8487 tttttttttt ttttctcctt tttttttcct ctttttttcc ttttctttcc tttggtggct 8547 ccatcttagc cctagtcacg gctagctgtg aaaggtccgt gagccgcttg actgcagaga 8607 gtgctgatac tggcctctct gcagatcaag t 8638

Claims

1. A host cell transfected with a self-replicating polynucleotide comprising:

(a) a 5′-Non Translated Region;

(b) a HCV polynucleotide coding region encoding an HCV polyprotein comprising: NS3, NS4A, NS4B, NS5A, and NS5B proteins, said polynucleotide coding region comprising one or more amino acid substitutions selected from the group consisting of: R(1135)K, S(1148)G, S(1560)G, K(1691)R, L(1701)F, I(1984)V, T(1993)A, G(2042)C, G(2042)R, S(2404)P, L(2155)P, P(2166)L, and m(2992)T, and

(c) a 3′-non translated region.

2. The host cell according to claim 1, wherein the host cell is a eukaryotic cell line.

3. The host cell according to claim 2, wherein said eukaryotic cell line is a hepatic cell line.

4. The host cell according to claim 3, wherein said hepatic cell line is Huh-7.

5. A RNA replication assay comprising the steps of:

(a) incubating the host cell according to claim 1 under conditions suitable for RNA replication;

(b) isolating the total cellular RNA from the cells; and

(c) analyzing the RNA so as to measure the amount of HCV RNA replicated.

6. The assay according to claim 5, wherein the analysis of RNA levels in step (c) is carried out by amplifying the RNA by real-time RT-PCR analysis using HCV specific primers so as to measure the amount of HCV RNA replicated.

7. The assay according to claim 5, wherein said polynucleotide encodes for a reporter gene, and the analysis of RNA levels in step (c) is carried out by assessing the level of reporter expressed.

8. A method for testing a compound for inhibiting HCV replication, including the steps of:

(a) carrying step (a) according to claim 5, in the presence or absence of the compound;

(b) isolating the total cellular RNA from the cells;

(c) analyzing the RNA so as to measure the amount of HCV RNA replicated; and

(d) comparing the levels of HCV RNA in cells in the absence and presence of the inhibitor,

wherein reduced RNA levels is indicative of the ability of the compound to inhibit replication.

9. The method according to claim 8, wherein said cell line is incubated with the test compound for about 3-4 days at a temperature of about 37° C.