质粒名称: | pBS NL4-3 envFS eGFP PR-D25A |
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出品公司: | Addgene |
目录编号: | 101334 |
存储实验室: | Jeremy Luban |
载体骨架基本信息 |
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载体名称: | pBluescript |
载体出品公司: | -- |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 慢病毒载体 |
筛选标记: | -- |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | NL4-3 envFS |
物种来源: | -- |
插入大小: | -- |
基因突变: | Protease D25A |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | -- |
3’测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pBS NL4-3 envFS eGFP PR-D25A was a gift from Jeremy Luban (Addgene plasmid # 101334 ; http://n2t.net/addgene:101334 ; RRID:Addgene_101334) |
参考文献部分: | Intron-containing RNA from the HIV-1 provirus activates type I interferon and inflammatory cytokines. McCauley SM, Kim K, Nowosielska A, Dauphin A, Yurkovetskiy L, Diehl WE, Luban J. Nat Commun. 2018 Dec 13;9(1):5305. doi: 10.1038/s41467-018-07753-2. 10.1038/s41467-018-07753-2 PubMed 30546110 |
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载体基本信息
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载体序列
LOCUS pBS NL4-3 envFS eGFP PR-D25A 12625 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION pBS NL4-3 envFS eGFP with mutation that disrupts Protease catalytic activity. ACCESSION . VERSION . KEYWORDS pBS NL4-3 envFS eGFP PR-D25A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12625) AUTHORS McCauley SM, Kim K, Nowosielska A, Dauphin A, Yurkovetskiy L, Diehl WE, Luban J TITLE Intron-containing RNA from the HIV-1 provirus activates type I interferon and inflammatory cytokines. JOURNAL Nat Commun. 2018 Dec 13;9(1):5305. doi: 10.1038/s41467-018-07753-2. PUBMED 30546110 REFERENCE 2 (bases 1 to 12625) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12625 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 377..634 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" CDS 790..2292 /codon_start=1 /gene="gag" /product="gag protein from human immunodeficiency virus 1" /label=HIV-1 gag /translation="MGARASVLSGGELDKWEKIRLRPGGKKQYKLKHIVWASRELERFA VNPGLLETSEGCRQILGQLQPSLQTGSEELRSLYNTIAVLYCVHQRIDVKDTKEALDKI EEEQNKSKKKAQQAAADTGNNSQVSQNYPIVQNLQGQMVHQAISPRTLNAWVKVVEEKA FSPEVIPMFSALSEGATPQDLNTMLNTVGGHQAAMQMLKETINEEAAEWDRLHPVHAGP IAPGQMREPRGSDIAGTTSTLQEQIGWMTHNPPIPVGEIYKRWIILGLNKIVRMYSPTS ILDIRQGPKEPFRDYVDRFYKTLRAEQASQEVKNWMTETLLVQNANPDCKTILKALGPG ATLEEMMTACQGVGGPGHKARVLAEAMSQVTNPATIMIQKGNFRNQRKTVKCFNCGKEG HIAKNCRAPRKKGCWKCGKEGHQMKDCTERQANFLGKIWPSHKGRPGNFLQSRPEPTAP PEESFRFGEETTTPSQKQEPIDKELYPLASLRSLFGSDPSSQ" CDS 2085..5096 /codon_start=1 /gene="pol" /product="pol protein from human immunodeficiency virus 1" /label=HIV-1 pol /translation="FFREDLAFPQGKAREFSSEQTRANSPTRRELQVWGRDNNSLSEAG ADRQGTVSFSFPQITLWQRPLVTIKIGGQLKEALLATGADDTVLEEMNLPGRWKPKMIG GIGGFIKVRQYDQILIEICGHKAIGTVLVGPTPVNIIGRNLLTQIGCTLNFPISPIETV PVKLKPGMDGPKVKQWPLTEEKIKALVEICTEMEKEGKISKIGPENPYNTPVFAIKKKD STKWRKLVDFRELNKRTQDFWEVQLGIPHPAGLKQKKSVTVLDVGDAYFSVPLDKDFRK YTAFTIPSINNETPGIRYQYNVLPQGWKGSPAIFQCSMTKILEPFRKQNPDIVIYQYMD DLYVGSDLEIGQHRTKIEELRQHLLRWGFTTPDKKHQKEPPFLWMGYELHPDKWTVQPI VLPEKDSWTVNDIQKLVGKLNWASQIYAGIKVRQLCKLLRGTKALTEVVPLTEEAELEL AENREILKEPVHGVYYDPSKDLIAEIQKQGQGQWTYQIYQEPFKNLKTGKYARMKGAHT NDVKQLTEAVQKIATESIVIWGKTPKFKLPIQKETWEAWWTEYWQATWIPEWEFVNTPP LVKLWYQLEKEPIIGAETFYVDGAANRETKLGKAGYVTDRGRQKVVPLTDTTNQKTELQ AIHLALQDSGLEVNIVTDSQYALGIIQAQPDKSESELVSQIIEQLIKKEKVYLAWVPAH KGIGGNEQVDKLVSAGIRKVLFLDGIDKAQEEHEKYHSNWRAMASDFNLPPVVAKEIVA SCDKCQLKGEAMHGQVDCSPGIWQLDCTHLEGKVILVAVHVASGYIEAEVIPAETGQET AYFLLKLAGRWPVKTVHTDNGSNFTSTTVKAACWWAGIKQEFGIPYNPQSQGVIESMNK ELKKIIGQVRDQAEHLKTAVQMAVFIHNFKRKGGIGGYSAGERIVDIIATDIQTKELQK QITKIQNFRVYYRDSRDPVWKGPAKLLWKGEGAVVIQDNSDIKVVPRRKAKIIRDYGKQ MAGDDCVASRQDED" misc_feature 4781..4898 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" misc_feature 7761..7994 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8179..8223 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" CDS 8517..8543 /codon_start=1 /product="nuclear export signal from the HIV Rev protein (Fischer et al., 1995)" /label=NES /translation="LPPLERLTL" CDS 8810..9526 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" CDS 8810..9523 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(8852..8873) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(9113..9132) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 9460..9481 /label=EGFP-C /note="EGFP, forward primer" LTR 9716..10349 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" rep_origin complement(10425..11013) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(10505..10524) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(11184..12044) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 11807..11826 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(12045..12149) /gene="bla" /label=AmpR promoter rep_origin 12176..12604 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(12263..12282) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 12473..12494 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 tggaagggct aatttggtcc caaaaaagac aagagatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tggcagaact acacaccagg gccagggatc agatatccac 121 tgacctttgg atggtgcttc aagttagtac cagttgaacc agagcaagta gaagaggcca 181 atgaaggaga gaacaacagc ttgttacacc ctatgagcca gcatgggatg gaggacccgg 241 agggagaagt attagtgtgg aagtttgaca gcctcctagc atttcgtcac atggcccgag 301 agctgcatcc ggagtactac aaagactgct gacatcgagc tttctacaag ggactttccg 361 ctggggactt tccagggagg tgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 gctacatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgctca aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagtaaa gccagaggag atctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcggta ttaagcgggg gagaattaga taaatgggaa 841 aaaattcggt taaggccagg gggaaagaaa caatataaac taaaacatat agtatgggca 901 agcagggagc tagaacgatt cgcagttaat cctggccttt tagagacatc agaaggctgt 961 agacaaatac tgggacagct acaaccatcc cttcagacag gatcagaaga acttagatca 1021 ttatataata caatagcagt cctctattgt gtgcatcaaa ggatagatgt aaaagacacc 1081 aaggaagcct tagataagat agaggaagag caaaacaaaa gtaagaaaaa ggcacagcaa 1141 gcagcagctg acacaggaaa caacagccag gtcagccaaa attaccctat agtgcagaac 1201 ctccaggggc aaatggtaca tcaggccata tcacctagaa ctttaaatgc atgggtaaaa 1261 gtagtagaag agaaggcttt cagcccagaa gtaataccca tgttttcagc attatcagaa 1321 ggagccaccc cacaagattt aaataccatg ctaaacacag tggggggaca tcaagcagcc 1381 atgcaaatgt taaaagagac catcaatgag gaagctgcag aatgggatag attgcatcca 1441 gtgcatgcag ggcctattgc accaggccag atgagagaac caaggggaag tgacatagca 1501 ggaactacta gtacccttca ggaacaaata ggatggatga cacataatcc acctatccca 1561 gtaggagaaa tctataaaag atggataatc ctgggattaa ataaaatagt aagaatgtat 1621 agccctacca gcattctgga cataagacaa ggaccaaagg aaccctttag agactatgta 1681 gaccgattct ataaaactct aagagccgag caagcttcac aagaggtaaa aaattggatg 1741 acagaaacct tgttggtcca aaatgcgaac ccagattgta agactatttt aaaagcattg 1801 ggaccaggag cgacactaga agaaatgatg acagcatgtc agggagtggg gggacccggc 1861 cataaagcaa gagttttggc tgaagcaatg agccaagtaa caaatccagc taccataatg 1921 atacagaaag gcaattttag gaaccaaaga aagactgtta agtgtttcaa ttgtggcaaa 1981 gaagggcaca tagccaaaaa ttgcagggcc cctaggaaaa agggctgttg gaaatgtgga 2041 aaggaaggac accaaatgaa agattgtact gagagacagg ctaatttttt agggaagatc 2101 tggccttccc acaagggaag gccagggaat tttcttcaga gcagaccaga gccaacagcc 2161 ccaccagaag agagcttcag gtttggggaa gagacaacaa ctccctctca gaagcaggag 2221 ccgatagaca aggaactgta tcctttagct tccctcagat cactctttgg cagcgacccc 2281 tcgtcacaat aaagataggg gggcaattaa aggaagctct attagctaca ggagcagatg 2341 atacagtatt agaagaaatg aatttgccag gaagatggaa accaaaaatg atagggggaa 2401 ttggaggttt tatcaaagta agacagtatg atcagatact catagaaatc tgcggacata 2461 aagctatagg tacagtatta gtaggaccta cacctgtcaa cataattgga agaaatctgt 2521 tgactcagat tggctgcact ttaaattttc ccattagtcc tattgagact gtaccagtaa 2581 aattaaagcc aggaatggat ggcccaaaag ttaaacaatg gccattgaca gaagaaaaaa 2641 taaaagcatt agtagaaatt tgtacagaaa tggaaaagga aggaaaaatt tcaaaaattg 2701 ggcctgaaaa tccatacaat actccagtat ttgccataaa gaaaaaagac agtactaaat 2761 ggagaaaatt agtagatttc agagaactta ataagagaac tcaagatttc tgggaagttc 2821 aattaggaat accacatcct gcagggttaa aacagaaaaa atcagtaaca gtactggatg 2881 tgggcgatgc atatttttca gttcccttag ataaagactt caggaagtat actgcattta 2941 ccatacctag tataaacaat gagacaccag ggattagata tcagtacaat gtgcttccac 3001 agggatggaa aggatcacca gcaatattcc agtgtagcat gacaaaaatc ttagagcctt 3061 ttagaaaaca aaatccagac atagtcatct atcaatacat ggatgatttg tatgtaggat 3121 ctgacttaga aatagggcag catagaacaa aaatagagga actgagacaa catctgttga 3181 ggtggggatt taccacacca gacaaaaaac atcagaaaga acctccattc ctttggatgg 3241 gttatgaact ccatcctgat aaatggacag tacagcctat agtgctgcca gaaaaggaca 3301 gctggactgt caatgacata cagaaattag tgggaaaatt gaattgggca agtcagattt 3361 atgcagggat taaagtaagg caattatgta aacttcttag gggaaccaaa gcactaacag 3421 aagtagtacc actaacagaa gaagcagagc tagaactggc agaaaacagg gagattctaa 3481 aagaaccggt acatggagtg tattatgacc catcaaaaga cttaatagca gaaatacaga 3541 agcaggggca aggccaatgg acatatcaaa tttatcaaga gccatttaaa aatctgaaaa 3601 caggaaagta tgcaagaatg aagggtgccc acactaatga tgtgaaacaa ttaacagagg 3661 cagtacaaaa aatagccaca gaaagcatag taatatgggg aaagactcct aaatttaaat 3721 tacccataca aaaggaaaca tgggaagcat ggtggacaga gtattggcaa gccacctgga 3781 ttcctgagtg ggagtttgtc aatacccctc ccttagtgaa gttatggtac cagttagaga 3841 aagaacccat aataggagca gaaactttct atgtagatgg ggcagccaat agggaaacta 3901 aattaggaaa agcaggatat gtaactgaca gaggaagaca aaaagttgtc cccctaacgg 3961 acacaacaaa tcagaagact gagttacaag caattcatct agctttgcag gattcgggat 4021 tagaagtaaa catagtgaca gactcacaat atgcattggg aatcattcaa gcacaaccag 4081 ataagagtga atcagagtta gtcagtcaaa taatagagca gttaataaaa aaggaaaaag 4141 tctacctggc atgggtacca gcacacaaag gaattggagg aaatgaacaa gtagataaat 4201 tggtcagtgc tggaatcagg aaagtactat ttttagatgg aatagataag gcccaagaag 4261 aacatgagaa atatcacagt aattggagag caatggctag tgattttaac ctaccacctg 4321 tagtagcaaa agaaatagta gccagctgtg ataaatgtca gctaaaaggg gaagccatgc 4381 atggacaagt agactgtagc ccaggaatat ggcagctaga ttgtacacat ttagaaggaa 4441 aagttatctt ggtagcagtt catgtagcca gtggatatat agaagcagaa gtaattccag 4501 cagagacagg gcaagaaaca gcatacttcc tcttaaaatt agcaggaaga tggccagtaa 4561 aaacagtaca tacagacaat ggcagcaatt tcaccagtac tacagttaag gccgcctgtt 4621 ggtgggcggg gatcaagcag gaatttggca ttccctacaa tccccaaagt caaggagtaa 4681 tagaatctat gaataaagaa ttaaagaaaa ttataggaca ggtaagagat caggctgaac 4741 atcttaagac agcagtacaa atggcagtat tcatccacaa ttttaaaaga aaagggggga 4801 ttggggggta cagtgcaggg gaaagaatag tagacataat agcaacagac atacaaacta 4861 aagaattaca aaaacaaatt acaaaaattc aaaattttcg ggtttattac agggacagca 4921 gagatccagt ttggaaagga ccagcaaagc tcctctggaa aggtgaaggg gcagtagtaa 4981 tacaagataa tagtgacata aaagtagtgc caagaagaaa agcaaagatc atcagggatt 5041 atggaaaaca gatggcaggt gatgattgtg tggcaagtag acaggatgag gattaacaca 5101 tggaaaagat tagtaaaaca ccatatgtat atttcaagga aagctaagga ctggttttat 5161 agacatcact atgaaagtac taatccaaaa ataagttcag aagtacacat cccactaggg 5221 gatgctaaat tagtaataac aacatattgg ggtctgcata caggagaaag agactggcat 5281 ttgggtcagg gagtctccat agaatggagg aaaaagagat atagcacaca agtagaccct 5341 gacctagcag accaactaat tcatctgcac tattttgatt gtttttcaga atctgctata 5401 agaaatacca tattaggacg tatagttagt cctaggtgtg aatatcaagc aggacataac 5461 aaggtaggat ctctacagta cttggcacta gcagcattaa taaaaccaaa acagataaag 5521 ccacctttgc ctagtgttag gaaactgaca gaggacagat ggaacaagcc ccagaagacc 5581 aagggccaca gagggagcca tacaatgaat ggacactaga gcttttagag gaacttaaga 5641 gtgaagctgt tagacatttt cctaggatat ggctccataa cttaggacaa catatctatg 5701 aaacttacgg ggatacttgg gcaggagtgg aagccataat aagaattctg caacaactgc 5761 tgtttatcca tttcagaatt gggtgtcgac atagcagaat aggcgttact cgacagagga 5821 gagcaagaaa tggagccagt agatcctaga ctagagccct ggaagcatcc aggaagtcag 5881 cctaaaactg cttgtaccaa ttgctattgt aaaaagtgtt gctttcattg ccaagtttgt 5941 ttcatgacaa aagccttagg catctcctat ggcaggaaga agcggagaca gcgacgaaga 6001 gctcatcaga acagtcagac tcatcaagct tctctatcaa agcagtaagt agtacatgta 6061 atgcaaccta taatagtagc aatagtagca ttagtagtag caataataat agcaatagtt 6121 gtgtggtcca tagtaatcat agaatatagg aaaatattaa gacaaagaaa aatagacagg 6181 ttaattgata gactaataga aagagcagaa gacagtggca atgagagtga aggagaagta 6241 tcagcacttg tggagatggg ggtggaaatg gggcaccatg ctccttggga tattgatgat 6301 ctgtagtgct acagaaaaat tgtgggtcac cgtctattat ggggtacctg tgtggaagga 6361 agcaaccacc actctatttt gtgcatcaga tgctaaagca tatatgatac agaggtacat 6421 aatgtttggg ccacacatgc ctgtgtaccc acagacccca acccacaaga agtagtattg 6481 gtaaatgtga cagaaaattt taacatgtgg aaaaatgaca tggtagaaca gatgcatgag 6541 gatataatca gtttatggga tcaaagccta aagccatgtg taaaattaac cccactctgt 6601 gttagtttaa agtgcactga tttgaagaat gatactaata ccaatagtag tagcgggaga 6661 atgataatgg agaaaggaga gataaaaaac tgctctttca atatcagcac aagcataaga 6721 gataaggtgc agaaagaata tgcattcttt tataaacttg atatagtacc aatagataat 6781 accagctata ggttgataag ttgtaacacc tcagtcatta cacaggcctg tccaaaggta 6841 tcctttgagc caattcccat acattattgt gccccggctg gttttgcgat tctaaaatgt 6901 aataataaga cgttcaatgg aacaggacca tgtacaaatg tcagcacagt acaatgtaca 6961 catggaatca ggccagtagt atcaactcaa ctgctgttaa atggcagtct agcagaagaa 7021 gatgtagtaa ttagatctgc caatttcaca gacaatgcta aaaccataat agtacagctg 7081 aacacatctg tagaaattaa ttgtacaaga cccaacaaca atacaagaaa aagtatccgt 7141 atccagaggg gaccagggag agcatttgtt acaataggaa aaataggaaa tatgagacaa 7201 gcacattgta acattagtag agcaaaatgg aatgccactt taaaacagat agctagcaaa 7261 ttaagagaac aatttggaaa taataaaaca ataatcttta agcaatcctc aggaggggac 7321 ccagaaattg taacgcacag ttttaattgt ggaggggaat ttttctactg taattcaaca 7381 caactgttta atagtacttg gtttaatagt acttggagta ctgaagggtc aaataacact 7441 gaaggaagtg acacaatcac actcccatgc agaataaaac aatttataaa catgtggcag 7501 gaagtaggaa aagcaatgta tgcccctccc atcagtggac aaattagatg ttcatcaaat 7561 attactgggc tgctattaac aagagatggt ggtaataaca acaatgggtc cgagatcttc 7621 agacctggag gaggcgatat gagggacaat tggagaagtg aattatataa atataaagta 7681 gtaaaaattg aaccattagg agtagcaccc accaaggcaa agagaagagt ggtgcagaga 7741 gaaaaaagag cagtgggaat aggagctttg ttccttgggt tcttgggagc agcaggaagc 7801 actatgggcg cagcgtcaat gacgctgacg gtacaggcca gacaattatt gtctgatata 7861 gtgcagcagc agaacaattt gctgagggct attgaggcgc aacagcatct gttgcaactc 7921 acagtctggg gcatcaaaca gctccaggca agaatcctgg ctgtggaaag atacctaaag 7981 gatcaacagc tcctggggat ttggggttgc tctggaaaac tcatttgcac cactgctgtg 8041 ccttggaatg ctagttggag taataaatct ctggaacaga tttggaataa catgacctgg 8101 atggagtggg acagagaaat taacaattac acaagcttaa tacactcctt aattgaagaa 8161 tcgcaaaacc agcaagaaaa gaatgaacaa gaattattgg aattagataa atgggcaagt 8221 ttgtggaatt ggtttaacat aacaaattgg ctgtggtata taaaattatt cataatgata 8281 gtaggaggct tggtaggttt aagaatagtt tttgctgtac tttctatagt gaatagagtt 8341 aggcagggat attcaccatt atcgtttcag acccacctcc caatcccgag gggacccgac 8401 aggcccgaag gaatagaaga agaaggtgga gagagagaca gagacagatc cattcgatta 8461 gtgaacggat ccttagcact tatctgggac gatctgcgga gcctgtgcct cttcagctac 8521 caccgcttga gagacttact cttgattgta acgaggattg tggaacttct gggacgcagg 8581 gggtgggaag ccctcaaata ttggtggaat ctcctacaat attggagtca ggagctaaag 8641 aatagtgctg ttagcttgct caatgccaca gccatagcag tagctgaggg gacagatagg 8701 gttatagaag tagtacaagg agcttgtaga gctattcgcc acatacctag aagaataaga 8761 cagggcttgg aaaggatttt gctataagat gggtggcgcg gccgctctgg tgagcaaggg 8821 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 8881 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 8941 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct 9001 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt 9061 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 9121 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 9181 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 9241 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa 9301 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca 9361 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca 9421 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 9481 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaactcg agacctagaa 9541 aaacatggag caatcacaag tagcaataca gcagctaaca atgctgcttg tgcctggcta 9601 gaagcacaag aggaggaaga ggtgggtttt ccagtcacac ctcaggtacc tttaagacca 9661 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa 9721 gggctaattc actcccaaag aagacaagat atccttgatc tgtggatcta ccacacacaa 9781 ggctacttcc ctgattggca gaactacaca ccagggccag gggtcagata tccactgacc 9841 tttggatggt gctacaagct agtaccagtt gagccagata aggtagaaga ggccaataaa 9901 ggagagaaca ccagcttgtt acaccctgtg agcctgcatg gaatggatga ccctgagaga 9961 gaagtgttag agtggaggtt tgacagccgc ctagcatttc atcacgtggc ccgagagctg 10021 catccggagt acttcaagaa ctgctgacat cgagcttgct acaagggact ttccgctggg 10081 gactttccag ggaggcgtgg cctgggcggg actggggagt ggcgagccct cagatgctgc 10141 atataagcag ctgctttttg cctgtactgg gtctctctgg ttagaccaga tctgagcctg 10201 ggagctctct ggctaactag ggaacccact gcttaagcct caataaagct tgccttgagt 10261 gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt aactagagat ccctcagacc 10321 cttttagtca gtgtggaaaa tctctagcag gccggcccaa ttgacatgtg agcaaaaggc 10381 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc 10441 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga 10501 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc 10561 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat 10621 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg 10681 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc 10741 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga 10801 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact 10861 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt 10921 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag 10981 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg 11041 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa 11101 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata 11161 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg 11221 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata 11281 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg 11341 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct 11401 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt 11461 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc 11521 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga 11581 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt 11641 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc 11701 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa 11761 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca 11821 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca 11881 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct 11941 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc 12001 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa 12061 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt 12121 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg 12181 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca 12241 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc 12301 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct 12361 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg 12421 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc 12481 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg 12541 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg 12601 aattttaaca aaatatttta attaa //
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