质粒名称: | pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro |
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出品公司: | Addgene |
目录编号: | 141385 |
存储实验室: | Nevan Krogan |
载体骨架基本信息 |
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载体名称: | pLVX-EF1alpha-IRES-Puro |
载体出品公司: | Takara Bio |
空载体大小: | 8825 bp |
完整质粒大小: | 9138 bp |
载体类型: | 哺乳细胞表达, Lentiviral |
筛选标记: | Puromycin |
载体抗性: | 氨苄霉素 |
生长温度: | 30 ℃ |
宿主菌: | NEB Stable |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | SARS-CoV-2-E |
物种来源: | SARS-CoV-2 |
基因突变: | human codon optimized |
启动子: | EF1alpha |
标签/融合蛋白: | 2xStrep (C terminal on insert) |
克隆方法: | 未知 |
文献引用方法 |
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材料和方法部分: |
pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro was a gift from Nevan Krogan (Addgene plasmid # 141385 ; http://n2t.net/addgene:141385 ; RRID:Addgene_141385)
For viral preps, please replace (Addgene plasmid # 141385) in the above sentence with: (Addgene viral prep # 141385-LV)
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参考文献部分: | A SARS-CoV-2 protein interaction map reveals targets for drug repurposing. Gordon DE, Jang GM, Bouhaddou M, Xu J, Obernier K, White KM, O'Meara MJ, Rezelj VV, Guo JZ, Swaney DL, Tummino TA, Huttenhain R, Kaake RM, Richards AL, Tutuncuoglu B, Foussard H, Batra J, Haas K, Modak M, Kim M, Haas P, Polacco BJ, Braberg H, Fabius JM, Eckhardt M, Soucheray M, Bennett MJ, Cakir M, McGregor MJ, Li Q, Meyer B, Roesch F, Vallet T, Mac Kain A, Miorin L, Moreno E, Naing ZZC, Zhou Y, Peng S, Shi Y, Zhang Z, Shen W, Kirby IT, Melnyk JE, Chorba JS, Lou K, Dai SA, Barrio-Hernandez I, Memon D, Hernandez-Armenta C, Lyu J, Mathy CJP, Perica T, Pilla KB, Ganesan SJ, Saltzberg DJ, Rakesh R, Liu X, Rosenthal SB, Calviello L, Venkataramanan S, Liboy-Lugo J, Lin Y, Huang XP, Liu Y, Wankowicz SA, Bohn M, Safari M, Ugur FS, Koh C, Savar NS, Tran QD, Shengjuler D, Fletcher SJ, O'Neal MC, Cai Y, Chang JCJ, Broadhurst DJ, Klippsten S, Sharp PP, Wenzell NA, Kuzuoglu-Ozturk D, Wang HY, Trenker R, Young JM, Cavero DA, Hiatt J, Roth TL, Rathore U, Subramanian A, Noack J, Hubert M, Stroud RM, Frankel AD, Rosenberg OS, Verba KA, Agard DA, Ott M, Emerman M, Jura N, von Zastrow M, Verdin E, Ashworth A, Schwartz O, d'Enfert C, Mukherjee S, Jacobson M, Malik HS, Fujimori DG, Ideker T, Craik CS, Floor SN, Fraser JS, Gross JD, Sali A, Roth BL, Ruggero D, Taunton J, Kortemme T, Beltrao P, Vignuzzi M, Garcia-Sastre A, Shokat KM, Shoichet BK, Krogan NJ. Nature. 2020 Jul;583(7816):459-468. doi: 10.1038/s41586-020-2286-9. Epub 2020 Apr 30. 10.1038/s41586-020-2286-9 PubMed 32353859 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro 9138 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Lentiviral expression of SARS-CoV-2 envelope (E) protein; transient expression and generate lentivirus. ACCESSION . VERSION . KEYWORDS pLVX-EF1alpha-SARS-CoV-2-E-2xStrep-IRES-Puro SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9138) AUTHORS Gordon DE, et al. TITLE A SARS-CoV-2-Human Protein-Protein Interaction Map Reveals Drug Targets and Potential Drug Repurposing JOURNAL BioRxiv March 2020 REFERENCE 2 (bases 1 to 9138) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..9138 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 1..1182 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 231..1173 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1130..1150 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1456..1479 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" CDS 1516..1539 /codon_start=1 /product="peptide that binds Strep-Tactin(R), an engineered form of streptavidin" /label=Strep-Tag II /translation="WSHPQFEK" misc_feature 1551..2124 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(1718..1735) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 1945..1964 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2157..2756 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(2157..2176) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 2653..2673 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 2770..3358 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2823..2843) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3241..3252) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 3565..4198 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(4326..4342) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4326..4342) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4339..4361) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4350..4366 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4374..4404) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4419..4440 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4557..4574) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4728..5316) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4808..4827) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5487..6347) /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 6110..6129 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6348..6452) /gene="bla" /label=AmpR promoter polyA_signal 6500..6634 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6537..6556) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6591..6610 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" LTR 6802..7435 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 7482..7607 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 8104..8337 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 8522..8566 /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" misc_feature 8829..8944 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" ORIGIN 1 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg 61 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg 121 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag 181 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg 241 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta 301 cttccacgcc cctggctgca gtacgtgatt cttgatcccg agcttcgggt tggaagtggg 361 tgggagagtt cgaggccttg cgcttaagga gccccttcgc ctcgtgcttg agttgaggcc 421 tggcttgggc gctggggccg ccgcgtgcga atctggtggc accttcgcgc ctgtctcgct 481 gctttcgata agtctctagc catttaaaat ttttgatgac ctgctgcgac gctttttttc 541 tggcaagata gtcttgtaaa tgcgggccaa gatctgcaca ctggtatttc ggtttttggg 601 gccgcgggcg gcgacggggc ccgtgcgtcc cagcgcacat gttcggcgag gcggggcctg 661 cgagcgcggc caccgagaat cggacggggg tagtctcaag ctggccggcc tgctctggtg 721 cctggcctcg cgccgccgtg tatcgccccg ccctgggcgg caaggctggc ccggtcggca 781 ccagttgcgt gagcggaaag atggccgctt cccggccctg ctgcagggag ctcaaaatgg 841 aggacgcggc gctcgggaga gcgggcgggt gagtcaccca cacaaaggaa aagggccttt 901 ccgtcctcag ccgtcgcttc atgtgactcc acggagtacc gggcgccgtc caggcacctc 961 gattagttct cgagcttttg gagtacgtcg tctttaggtt ggggggaggg gttttatgcg 1021 atggagtttc cccacactga gtgggtggag actgaagtta ggccagcttg gcacttgatg 1081 taattctcct tggaatttgc cctttttgag tttggatctt ggttcattct caagcctcag 1141 acagtggttc aaagtttttt tcttccattt caggtgtcgt gaggatctat ttccggtgaa 1201 ttcgccgcca ccatgtacag cttcgtatca gaagaaaccg ggacactgat cgtaaattct 1261 gtgctcttgt ttctggcatt cgtcgtattt ctcctcgtca cactggcaat tctgactgca 1321 ttgaggcttt gcgcctactg ttgtaacatt gtcaatgtat ctctcgtgaa accctcattc 1381 tacgtttaca gcagggtgaa gaatctcaat tctagcaggg tgccggatct cctcgttctc 1441 gaaggcggcg ggggatggag ccatccacaa ttcgagaaag gcggtggttc aggaggaggt 1501 agcgggggtg gatcatggtc acatccgcag tttgaaaagt aaggatcccg cccctctccc 1561 tccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg tgcgtttgtc 1621 tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg gaaacctggc 1681 cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg aatgcaaggt 1741 ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca aacaacgtct 1801 gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct ctgcggccaa 1861 aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca cgttgtgagt 1921 tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa ggggctgaag 1981 gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg cacatgcttt 2041 acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg ggacgtggtt 2101 ttcctttgaa aaacacgatg ataagcttgc cacaacccac aaggagacga ccttccatga 2161 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtcccccgg gccgtacgca 2221 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgac ccggaccgcc 2281 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg 2341 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg 2401 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc 2461 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg 2521 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg 2581 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga 2641 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg 2701 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctagacgc 2761 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac 2821 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt 2881 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat 2941 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca 3001 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc 3061 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg 3121 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca 3181 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct 3241 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt 3301 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct 3361 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag 3421 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc 3481 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc 3541 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata 3601 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac 3661 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg 3721 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga 3781 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc 3841 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc 3901 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga 3961 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg 4021 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg 4081 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt 4141 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt 4201 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag 4261 agtgagaggc cttgacattg ctagcgttta ccgtcgacct ctagctagag cttggcgtaa 4321 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4381 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4441 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 4501 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 4561 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 4621 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 4681 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 4741 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 4801 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 4861 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 4921 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 4981 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 5041 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 5101 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 5161 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 5221 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 5281 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 5341 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 5401 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 5461 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 5521 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 5581 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 5641 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 5701 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 5761 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 5821 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 5881 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 5941 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 6001 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 6061 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 6121 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 6181 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 6241 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 6301 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 6361 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 6421 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 6481 gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca aataaagcaa 6541 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 6601 caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag ctaaccaaaa 6661 tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt ggtttcattt 6721 actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta tttgttaaat 6781 atgtactaca aacttagtag ttggaagggc taattcactc ccaaagaaga caagatatcc 6841 ttgatctgtg gatctaccac acacaaggct acttccctga ttagcagaac tacacaccag 6901 ggccaggggt cagatatcca ctgacctttg gatggtgcta caagctagta ccagttgagc 6961 cagataaggt agaagaggcc aataaaggag agaacaccag cttgttacac cctgtgagcc 7021 tgcatgggat ggatgacccg gagagagaag tgttagagtg gaggtttgac agccgcctag 7081 catttcatca cgtggcccga gagctgcatc cggagtactt caagaactgc tgatatcgag 7141 cttgctacaa gggactttcc gctggggact ttccagggag gcgtggcctg ggcgggactg 7201 gggagtggcg agccctcaga tcctgcatat aagcagctgc tttttgcctg tactgggtct 7261 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 7321 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 7381 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 7441 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 7501 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 7561 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 7621 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 7681 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 7741 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 7801 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 7861 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 7921 aaagtaagac caccgcacag caagcggccg gccgctgatc ttcagacctg gaggaggaga 7981 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt 8041 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg 8101 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc 8161 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa 8221 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa 8281 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg 8341 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg 8401 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga 8461 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga 8521 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa 8581 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg 8641 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc 8701 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga 8761 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatctcgacg 8821 gtatcgcctt taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag 8881 acataatagc aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa 8941 attttcgggt ttattacagg gacagcagag atccagttta tcgatgagta attcatacaa 9001 aaggactcgc ccctgccttg gggaatccca gggaccgtcg ttaaactccc actaacgtag 9061 aacccagaga tcgctgcgtt cccgccccct cacccgcccg ctctcgtcat cactgaggtg 9121 gagaagagca tgcgtgag //
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