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[1]
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NUCLEOTIDE SEQUENCE [GENOMIC RNA].
DOI=10.1016/0042-6822(84)90428-8; PubMed=6322438 [NCBI, ExPASy, EBI, Israel, Japan]
Strauss E.G.,
Rice C.M.,
Strauss J.H.;
"Complete nucleotide sequence of the genomic RNA of Sindbis virus.";
Virology 133:92-110(1984).
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[2]
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NUCLEOTIDE SEQUENCE [GENOMIC RNA] OF 1-54.
DOI=10.1016/S0022-2836(83)80319-2; PubMed=6308269 [NCBI, ExPASy, EBI, Israel, Japan]
Ou J.H.,
Strauss E.G.,
Strauss J.H.;
"The 5'-terminal sequences of the genomic RNAs of several alphaviruses.";
J. Mol. Biol. 168:1-15(1983).
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[3]
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NUCLEOTIDE SEQUENCE [GENOMIC RNA] OF 1429-2512.
PubMed=6577423 [NCBI, ExPASy, EBI, Israel, Japan]
Strauss E.G.,
Rice C.M.,
Strauss J.H.;
"Sequence coding for the alphavirus nonstructural proteins is interrupted by an opal termination codon.";
Proc. Natl. Acad. Sci. U.S.A. 80:5271-5275(1983).
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[4]
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NUCLEOTIDE SEQUENCE [GENOMIC RNA] OF 2431-2512.
PubMed=6291034 [NCBI, ExPASy, EBI, Israel, Japan]
Ou J.-H.,
Rice C.M.,
Dalgarno L.,
Strauss E.G.,
Strauss J.H.;
"Sequence studies of several alphavirus genomic RNAs in the region containing the start of the subgenomic RNA.";
Proc. Natl. Acad. Sci. U.S.A. 79:5235-5239(1982).
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[5]
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SUBCELLULAR LOCATION OF NON-STRUCTURAL PROTEINS.
DOI=10.1083/jcb.107.6.2075; PubMed=2904446 [NCBI, ExPASy, EBI, Israel, Japan]
Froshauer S.,
Kartenbeck J.,
Helenius A.;
"Alphavirus RNA replicase is located on the cytoplasmic surface of endosomes and lysosomes.";
J. Cell Biol. 107:2075-2086(1988).
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[6]
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PROTEOLYTIC PROCESSING OF POLYPROTEIN.
PubMed=2529379 [NCBI, ExPASy, EBI, Israel, Japan]
Hardy W.R.,
Strauss J.H.;
"Processing the nonstructural polyproteins of sindbis virus: nonstructural proteinase is in the C-terminal half of nsP2 and functions both in cis and in trans.";
J. Virol. 63:4653-4664(1989).
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[7]
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MUTAGENESIS OF OPAL STOP CODON BETWEEN TYR-1896 AND LEU-1897.
PubMed=2521676 [NCBI, ExPASy, EBI, Israel, Japan]
Li G.P.,
Rice C.M.;
"Mutagenesis of the in-frame opal termination codon preceding nsP4 of Sindbis virus: studies of translational readthrough and its effect on virus replication.";
J. Virol. 63:1326-1337(1989).
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[8]
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PROTEOLYTIC PROCESSING OF POLYPROTEIN BY NSP2.
PubMed=2142454 [NCBI, ExPASy, EBI, Israel, Japan]
de Groot R.J.,
Hardy W.R.,
Shirako Y.,
Strauss J.H.;
"Cleavage-site preferences of Sindbis virus polyproteins containing the non-structural proteinase. Evidence for temporal regulation of polyprotein processing in vivo.";
EMBO J. 9:2631-2638(1990).
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[9]
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UBIQUITIN DEGRADATION OF NSP4.
PubMed=1924357 [NCBI, ExPASy, EBI, Israel, Japan]
de Groot R.J.,
Ruemenapf T.,
Kuhn R.J.,
Strauss E.G.,
Strauss J.H.;
"Sindbis virus RNA polymerase is degraded by the N-end rule pathway.";
Proc. Natl. Acad. Sci. U.S.A. 88:8967-8971(1991).
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[10]
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MUTAGENESIS OF CYS-1021; HIS-1098 AND TRP-1099.
DOI=10.1016/0042-6822(92)90268-T; PubMed=1448929 [NCBI, ExPASy, EBI, Israel, Japan]
Strauss E.G.,
De Groot R.J.,
Levinson R.,
Strauss J.H.;
"Identification of the active site residues in the nsP2 proteinase of Sindbis virus.";
Virology 191:932-940(1992).
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[11]
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FUNCTION.
PubMed=7517863 [NCBI, ExPASy, EBI, Israel, Japan]
Lemm J.A.,
Ruemenapf T.,
Strauss E.G.,
Strauss J.H.,
Rice C.M.;
"Polypeptide requirements for assembly of functional Sindbis virus replication complexes: a model for the temporal regulation of minus- and plus-strand RNA synthesis.";
EMBO J. 13:2925-2934(1994).
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[12]
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FUNCTION OF P123.
PubMed=8107248 [NCBI, ExPASy, EBI, Israel, Japan]
Shirako Y.,
Strauss J.H.;
"Regulation of Sindbis virus RNA replication: uncleaved P123 and nsP4 function in minus-strand RNA synthesis, whereas cleaved products from P123 are required for efficient plus-strand RNA synthesis.";
J. Virol. 68:1874-1885(1994).
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[13]
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MUTAGENESIS OF HIS-39; HIS-81; ASP-91; ARG-94; TYR-249 AND ILE-369.
DOI=10.1006/viro.1996.0147; PubMed=8610444 [NCBI, ExPASy, EBI, Israel, Japan]
Wang H.-L.,
O'Rear J.,
Stollar V.;
"Mutagenesis of the Sindbis virus nsP1 protein: effects on methyltransferase activity and viral infectivity.";
Virology 217:527-531(1996).
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[14]
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MUTAGENESIS OF TYR-1903.
PubMed=9499091 [NCBI, ExPASy, EBI, Israel, Japan]
Shirako Y.,
Strauss J.H.;
"Requirement for an aromatic amino acid or histidine at the N-terminus of Sindbis virus RNA polymerase.";
J. Virol. 72:2310-2315(1998).
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[15]
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PALMITOYLATION AT CYS-420, AND MUTAGENESIS OF CYS-420.
DOI=10.1128/JVI.74.15.6725-6733.2000; PubMed=10888610 [NCBI, ExPASy, EBI, Israel, Japan]
Ahola T.,
Kujala P.,
Tuittila M.,
Blom T.,
Laakkonen P.,
Hinkkanen A.,
Auvinen P.;
"Effects of palmitoylation of replicase protein nsP1 on alphavirus infection.";
J. Virol. 74:6725-6733(2000).
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[16]
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INDUCTION.
DOI=10.1101/gad.357006; PubMed=16391235 [NCBI, ExPASy, EBI, Israel, Japan]
Ventoso I.,
Sanz M.A.,
Molina S.,
Berlanga J.J.,
Carrasco L.,
Esteban M.;
"Translational resistance of late alphavirus mRNA to eIF2alpha phosphorylation: a strategy to overcome the antiviral effect of protein kinase PKR.";
Genes Dev. 20:87-100(2006).
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- FUNCTION: P123 and P123' are short-lived polyproteins, accumulating during early stage of infection. P123 is directly translated from the genome, whereas P123' is a product of the cleavage of P1234. They localize the viral replication complex to the cytoplasmic surface of modified endosomes and lysosomes. By interacting with nsP4, they start viral genome replication into antigenome. After these early events, P123 and P123' are cleaved sequentially into nsP1, nsP2 and nsP3/nsP3'. This sequence of delayed processing would allow correct assembly and membrane association of the RNA polymerase complex.
- FUNCTION: nsP1 is a cytoplasmic capping enzyme. This function is necessary since all viral RNAs are synthesized in the cytoplasm, and host capping enzymes are restricted to the nucleus. The enzymatic reaction involves a covalent link between 7-methyl-GMP and nsP1, whereas eukaryotic capping enzymes form a covalent complex only with GMP. nsP1 capping would consist in the following reactions: GTP is first methylated and then forms the m7GMp-nsP1 complex, from which 7-methyl-GMP complex is transferred to the mRNA to create the cap structure. Palmitoylated nsP1 is remodeling host cell cytoskeleton, and induces filopodium-like structure formation at the surface of the host cell.
- FUNCTION: nsP2 has two separate domain with different biological activities. The N-terminal section is part of the RNA polymerase complex and has RNA trisphosphatase and RNA helicase activity. The C-terminal section harbors a protease that specifically cleaves and releases the four mature proteins.
- FUNCTION: nsP3 and nsP3' are essential for minus strand and subgenomic 26S mRNA synthesis.
- FUNCTION: nsP4 is a RNA dependent RNA polymerase. It replicates genomic and antigenomic RNA by recognizing replications specific signals. Transcribes also a 26S subgenomic mRNA by initiating RNA synthesis internally on antigenomic RNA. This 26S mRNA encodes for structural proteins. nsP4 is a short-lived protein regulated by several ways: the opal codon readthrough and degradation by ubiquitin pathway.
- CATALYTIC ACTIVITY: S-adenosyl-L-methionine + GTP = m7GTP.
- CATALYTIC ACTIVITY: m7GTP + (5')pp-Pur-mRNA = diphosphate + m7G(5')ppp-Pur-mRNA.
- CATALYTIC ACTIVITY: (5')ppp-mRNA + H2O = (5')pp-mRNA + phosphate.
- CATALYTIC ACTIVITY: A 5'-phosphopolynucleotide + H2O = a polynucleotide + phosphate.
- CATALYTIC ACTIVITY: NTP + H2O = NDP + phosphate.
- CATALYTIC ACTIVITY: Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1).
- SUBUNIT: P123 interacts with nsP4; nsP1, nsP2, nsP3 and nsP4 interact with each other, and with uncharacterized host factors.
- SUBCELLULAR LOCATION: Non-structural polyprotein: Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side. Note=Located on the cytoplasmic surface of modified endosomes and lysosomes, also called cytopathic vacuoles type I (CPVI). These vacuoles contain numerous small circular invaginations (spherules) which may be the sites of RNA synthesis.
- SUBCELLULAR LOCATION: P123: Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side.
- SUBCELLULAR LOCATION: P123': Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side.
- SUBCELLULAR LOCATION: mRNA-capping enzyme nsP1: Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side. Cell membrane; Peripheral membrane protein; Cytoplasmic side. Cell projection, filopodium. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then a fraction of nsP1 localizes to the inner surface of the plasma membrane and its filopodial extensions.
- SUBCELLULAR LOCATION: Protease/triphosphatase/NTPase/helicase nsP2: Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side. Nucleus. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then approximately half of nsP2 is found in the nucleus.
- SUBCELLULAR LOCATION: Non-structural protein 3: Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side. Cytoplasm. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then nsP3 and nsP3' seems to aggregate in cytoplasm.
- SUBCELLULAR LOCATION: Non-structural protein 3': Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side. Cytoplasm. Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then nsP3 and nsP3' seems to aggregate in cytoplasm.
- SUBCELLULAR LOCATION: RNA-directed RNA polymerase nsP4: Endosome membrane; Peripheral membrane protein; Cytoplasmic side. Lysosome membrane; Peripheral membrane protein; Cytoplasmic side.
- INDUCTION: Viral replication produces dsRNA in the late phase of infection, resulting in a strong activation of host EIF2AK2/PKR, leading to almost complete phosphorylation of EIF2A. This inactivates completely cellular translation initiation, resulting in a dramatic shutoff of proteins synthesis. Translation of viral non-structural polyprotein and all cellular proteins are stopped in infected cell between 2 and 4 hours post infection. Only the 26S mRNA is still translated into viral structural proteins, presumably through a unique mechanism of enhancer element which counteract the translation inhibition mediated by EIF2A. By doing this, the virus uses the cellular defense for its own advantage: shutoff of cellular translation allows to produce big amounts of structural proteins needed for the virus to bud out of the doomed cell.
- PTM: Specific enzymatic cleavages in vivo yield mature proteins. The polyprotein is synthesized as P123, or P1234 by stop codon readthrough. These polyproteins are processed differently depending on the stage of infection. In early stages, P1234 is first cleaved in trans, through its nsP2 protease activity, releasing P123' and nsP4. P123/P123' and nsP4 start to replicate the viral genome into its antigenome. After these early events, nsP1 is cleaved in cis by nsP2 protease, releasing the P23/P23' polyprotein. Cleavage of nsP1 exposes an 'activator' at the N-terminus of P23/P23' which induces its cleavage into nsP2 and nsP3 by the viral protease. This sequence of delayed processing would allow correct assembly and membrane association of the RNA-polymerase complex. In the late stage of infection, the presence of free nsP2 in the cytoplasm cleaves P1234 quickly into P12 and P34, then into the four nsP.
- PTM: nsP1 is palmitoylated by host.
- PTM: nsP4 is ubiquitinated; targets the protein for rapid degradation via the ubiquitin system.
- MISCELLANEOUS: The genome encodes for P123, but readthrough of a terminator codon UGA occurs between the codons for Tyr-1896 and Leu-1897. This readthrough produces P1234, cleaved quickly by nsP2 into P123' and nsP4. Further processing of p123' gives nsP1, nsP2 and nsP3' which is 6 amino-acids longer than nsP3 since the cleavage site is after the readthrough. This unusual molecular mechanism ensures that few nsP4 are produced compared to other non-structural proteins. Mutant viruses with no alternative termination site grow significantly slower than wild-type virus.
- SIMILARITY: Contains 1 Macro domain.
- SIMILARITY: Contains 1 peptidase C9 domain [view classification].
- SIMILARITY: Contains 1 RdRp catalytic domain.
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Copyrighted by the UniProt Consortium, see http://www.uniprot.org/terms.
Distributed under the Creative Commons Attribution-NoDerivs License.
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| Length: 2512 AA [This is the length of the unprocessed precursor] |
Molecular weight: 279549 Da [This is the MW of the unprocessed precursor] |
CRC64: F3656FC8BB495726 [This is a checksum on the sequence] |
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10 20 30 40 50 60
MEKPVVNVDV DPQSPFVVQL QKSFPQFEVV AQQVTPNDHA NARAFSHLAS KLIELEVPTT
70 80 90 100 110 120
ATILDIGSAP ARRMFSEHQY HCVCPMRSPE DPDRMMKYAS KLAEKACKIT NKNLHEKIKD
130 140 150 160 170 180
LRTVLDTPDA ETPSLCFHND VTCNMRAEYS VMQDVYINAP GTIYHQAMKG VRTLYWIGFD
190 200 210 220 230 240
TTQFMFSAMA GSYPAYNTNW ADEKVLEARN IGLCSTKLSE GRTGKLSIMR KKELKPGSRV
250 260 270 280 290 300
YFSVGSTLYP EHRASLQSWH LPSVFHLNGK QSYTCRCDTV VSCEGYVVKK ITISPGITGE
310 320 330 340 350 360
TVGYAVTHNS EGFLLCKVTD TVKGERVSFP VCTYIPATIC DQMTGIMATD ISPDDAQKLL
370 380 390 400 410 420
VGLNQRIVIN GRTNRNTNTM QNYLLPIIAQ GFSKWAKERK DDLDNEKMLG TRERKLTYGC
430 440 450 460 470 480
LWAFRTKKVH SFYRPPGTQT CVKVPASFSA FPMSSVWTTS LPMSLRQKLK LALQPKKEEK
490 500 510 520 530 540
LLQVSEELVM EAKAAFEDAQ EEARAEKLRE ALPPLVADKG IEAAAEVVCE VEGLQADIGA
550 560 570 580 590 600
ALVETPRGHV RIIPQANDRM IGQYIVVSPN SVLKNAKLAP AHPLADQVKI ITHSGRSGRY
610 620 630 640 650 660
AVEPYDAKVL MPAGGAVPWP EFLALSESAT LVYNEREFVN RKLYHIAMHG PAKNTEEEQY
670 680 690 700 710 720
KVTKAELAET EYVFDVDKKR CVKKEEASGL VLSGELTNPP YHELALEGLK TRPAVPYKVE
730 740 750 760 770 780
TIGVIGTPGS GKSAIIKSTV TARDLVTSGK KENCREIEAD VLRLRGMQIT SKTVDSVMLN
790 800 810 820 830 840
GCHKAVEVLY VDEAFACHAG ALLALIAIVR PRKKVVLCGD PMQCGFFNMM QLKVHFNHPE
850 860 870 880 890 900
KDICTKTFYK YISRRCTQPV TAIVSTLHYD GKMKTTNPCK KNIEIDITGA TKPKPGDIIL
910 920 930 940 950 960
TCFRGWVKQL QIDYPGHEVM TAAASQGLTR KGVYAVRQKV NENPLYAITS EHVNVLLTRT
970 980 990 1000 1010 1020
EDRLVWKTLQ GDPWIKQPTN IPKGNFQATI EDWEAEHKGI IAAINSPTPR ANPFSCKTNV
1030 1040 1050 1060 1070 1080
CWAKALEPIL ATAGIVLTGC QWSELFPQFA DDKPHSAIYA LDVICIKFFG MDLTSGLFSK
1090 1100 1110 1120 1130 1140
QSIPLTYHPA DSARPVAHWD NSPGTRKYGY DHAIAAELSR RFPVFQLAGK GTQLDLQTGR
1150 1160 1170 1180 1190 1200
TRVISAQHNL VPVNRNLPHA LVPEYKEKQP GPVKKFLNQF KHHSVLVVSE EKIEAPRKRI
1210 1220 1230 1240 1250 1260
EWIAPIGIAG ADKNYNLAFG FPPQARYDLV FINIGTKYRN HHFQQCEDHA ATLKTLSRSA
1270 1280 1290 1300 1310 1320
LNCLNPGGTL VVKSYGYADR NSEDVVTALA RKFVRVSAAR PDCVSSNTEM YLIFRQLDNS
1330 1340 1350 1360 1370 1380
RTRQFTPHHL NCVISSVYEG TRDGVGAAPS YRTKRENIAD CQEEAVVNAA NPLGRPGEGV
1390 1400 1410 1420 1430 1440
CRAIYKRWPT SFTDSATETG TARMTVCLGK KVIHAVGPDF RKHPEAEALK LLQNAYHAVA
1450 1460 1470 1480 1490 1500
DLVNEHNIKS VAIPLLSTGI YAAGKDRLEV SLNCLTTALD RTDADVTIYC LDKKWKERID
1510 1520 1530 1540 1550 1560
AALQLKESVT ELKDEDMEID DELVWIHPDS CLKGRKGFST TKGKLYSYFE GTKFHQAAKD
1570 1580 1590 1600 1610 1620
MAEIKVLFPN DQESNEQLCA YILGETMEAI REKCPVDHNP SSSPPKTLPC LCMYAMTPER
1630 1640 1650 1660 1670 1680
VHRLRSNNVK EVTVCSSTPL PKHKIKNVQK VQCTKVVLFN PHTPAFVPAR KYIEVPEQPT
1690 1700 1710 1720 1730 1740
APPAQAEEAP EVVATPSPST ADNTSLDVTD ISLDMDDSSE GSLFSSFSGS DNSITSMDSW
1750 1760 1770 1780 1790 1800
SSGPSSLEIV DRRQVVVADV HAVQEPAPIP PPRLKKMARL AAARKEPTPP ASNSSESLHL
1810 1820 1830 1840 1850 1860
SFGGVSMSLG SIFDGETARQ AAVQPLATGP TDVPMSFGSF SDGEIDELSR RVTESEPVLF
1870 1880 1890 1900 1910 1920
GSFEPGEVNS IISSRSAVSF PLRKQRRRRR SRRTEYLTGV GGYIFSTDTG PGHLQKKSVL
1930 1940 1950 1960 1970 1980
QNQLTEPTLE RNVLERIHAP VLDTSKEEQL KLRYQMMPTE ANKSRYQSRK VENQKAITTE
1990 2000 2010 2020 2030 2040
RLLSGLRLYN SATDQPECYK ITYPKPLYSS SVPANYSDPQ FAVAVCNNYL HENYPTVASY
2050 2060 2070 2080 2090 2100
QITDEYDAYL DMVDGTVACL DTATFCPAKL RSYPKKHEYR APNIRSAVPS AMQNTLQNVL
2110 2120 2130 2140 2150 2160
IAATKRNCNV TQMRELPTLD SATFNVECFR KYACNDEYWE EFARKPIRIT TEFVTAYVAR
2170 2180 2190 2200 2210 2220
LKGPKAAALF AKTYNLVPLQ EVPMDRFVMD MKRDVKVTPG TKHTEERPKV QVIQAAEPLA
2230 2240 2250 2260 2270 2280
TAYLCGIHRE LVRRLTAVLL PNIHTLFDMS AEDFDAIIAE HFKQGDPVLE TDIASFDKSQ
2290 2300 2310 2320 2330 2340
DDAMALTGLM ILEDLGVDQP LLDLIECAFG EISSTHLPTG TRFKFGAMMK SGMFLTLFVN
2350 2360 2370 2380 2390 2400
TVLNVVIASR VLEERLKTSR CAAFIGDDNI IHGVVSDKEM AERCATWLNM EVKIIDAVIG
2410 2420 2430 2440 2450 2460
ERPPYFCGGF ILQDSVTSTA CRVADPLKRL FKLGKPLPAD DEQDEDRRRA LLDETKAWFR
2470 2480 2490 2500 2510
VGITGTLAVA VTTRYEVDNI TPVLLALRTF AQSKRAFQAI RGEIKHLYGG PK
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P03317 in FASTA format |
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