质粒名称: | pAAVS1-P-CAG-GFP |
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出品公司: | 爱迪基因 |
目录编号: | 80491 |
存储实验室: | Knut Woltjen |
载体骨架基本信息 |
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载体名称: | pCR-TOPO |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素和卡那霉素 |
空载体大小: | -- |
完整质粒大小: | 3000 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | 新霉素(G418) |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | CAG-GFP |
物种来源: | -- |
插入大小: | 2447 bp |
基因突变: | -- |
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: | AgeI(未破坏) |
3' 克隆位点: | NotI (未破坏) |
5' 测序引物 | CMV-F |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | pAAVS1-P-CAG-GFP was a gift from Knut Woltjen (Addgene plasmid # 80491 ; http://n2t.net/addgene:80491 ; RRID:Addgene_80491) |
参考文献部分: | A CRISPR-Cas9 System for Genetic Engineering of Filamentous Fungi. Nodvig CS, Nielsen JB, Kogle ME, Mortensen UH. PLoS One. 2015 Jul 15;10(7):e0133085. doi: 10.1371/journal.pone.0133085. eCollection 2015. PONE-D-15-11561 [pii] PubMed 26177455 |
买家导航
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 pAAVS1-P-CAG-GFP 9608 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION donor vector for AAVS1 targeting (puromycin selection) and constitutive GFP expression. ACCESSION . VERSION . KEYWORDS pAAVS1-P-CAG-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9608) AUTHORS Oceguera-Yanez F, Kim SI, Matsumoto T, Tan GW, Xiang L, Hatani T, Kondo T, Ikeya M, Yoshida Y, Inoue H, Woltjen K TITLE Engineering the AAVS1 locus for consistent and scalable transgene expression in human iPSCs and their differentiated derivatives. JOURNAL Methods. 2015 Dec 18. pii: S1046-2023(15)30181-X. doi: 10.1016/j.ymeth.2015.12.012. PUBMED 26707206 REFERENCE 2 (bases 1 to 9608) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9608 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /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)." primer_bind 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 242..260 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_feature 295..1098 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" misc_feature 1105..1130 /label=SA /note="splice acceptor site" CDS 1154..1207 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 1217..1816 /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(1217..1236) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 1713..1733 /label=Puro-F /note="Puromycin resistance gene, forward primer" primer_bind complement(1848..1865) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 1854..2078 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 2082..2461 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2464..2739 /label=chicken beta-actin promoter intron 2740..3748 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3672..3694 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3756..3775 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" regulatory 3803..3812 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3809..4528 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3854..3875) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(4115..4134) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 4462..4483 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(4612..4631) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 4677..4732 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(4731..4750) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" misc_feature 5104..5940 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" primer_bind complement(5980..5999) /label=T7 /note="T7 promoter, forward primer" promoter complement(5981..5999) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6006..6023) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6006..6022) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6015..6037) /label=M13/pUC Forward /note="In lacZ gene" CDS 6160..6462 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 6341..6360 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 6811..7605 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6865..6884) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7475..7494 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS complement(7855..8715) /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 8478..8497 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 8839..9427 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9328..9347 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 9581..9598 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca 241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt 301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg 361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct 421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc 481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg 541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc 601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg 661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt 721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt 781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg 841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca 901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat 961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc 1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc 1081 tccaccccac agtggggcaa gcttctgacc tcttctcttc ctcccacagg gcctcgagag 1141 atctggcagc ggagagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc 1201 cggccctagg ctcgagatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga 1261 cgtccccagg gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca 1321 caccgtcgat ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac 1381 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt 1441 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat 1501 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc 1561 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca 1621 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg 1681 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg 1741 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg 1801 caagcccggt gcctgatcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc 1861 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg 1921 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag 1981 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga 2041 caatagcagg catgctgggg atgcggtggg ctctatgggt cgacattgat tattgactag 2101 ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt 2161 tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac 2221 gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg 2281 ggtggactat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag 2341 tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat 2401 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat 2461 gggtcgaggt gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc 2521 caattttgta tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg 2581 gggcgcgcgc caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt 2641 gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg 2701 cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gttgccttcg 2761 ccccgtgccc cgctccgcgc cgcctcgcgc cgcccgcccc ggctctgact gaccgcgtta 2821 ctcccacagg tgagcgggcg ggacggccct tctcctccgg gctgtaatta gcgcttggtt 2881 taatgacggc tcgtttcttt tctgtggctg cgtgaaagcc ttaaagggct ccgggagggc 2941 cctttgtgcg ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc 3001 cgcgtgcggc ccgcgctgcc cggcggctgt gagcgctgcg ggcgcggcgc ggggctttgt 3061 gcgctccgcg tgtgcgcgag gggagcgcgg ccgggggcgg tgccccgcgg tgcggggggg 3121 ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag cagggggtgt 3181 gggcgcggcg gtcgggctgt aacccccccc tgcacccccc tccccgagtt gctgagcacg 3241 gcccggcttc gggtgcgggg ctccgtgcgg ggcgtggcgc ggggctcgcc gtgccgggcg 3301 gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc gcctcgggcc ggggagggct 3361 cgggggaggg gcgcggcggc cccggagcgc cggcggctgt cgaggcgcgg cgagccgcag 3421 ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt cccaaatctg 3481 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg gcgaagcggt 3541 gcggcgccgg caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc 3601 cccttctcca tctccagcct cggggctgcc gcagggggac ggctgccttc gggggggacg 3661 gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct ctgctaacca 3721 tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta ttgtgctgtc 3781 tcatcatttt ggcaaagaat tcgccaccat ggtgagcaag ggcgaggagc tgttcaccgg 3841 ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc 3901 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac 3961 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg 4021 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga 4081 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc 4141 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt 4201 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt 4261 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa 4321 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga 4381 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga 4441 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac 4501 tctcggcatg gacgagctgt acaagtaaga attcactcct caggtgcagg ctgcctatca 4561 gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc 4621 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata 4681 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag 4741 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc 4801 aacatatgcc atatgctggc tgccatgaac aaaggtggct ataaagaggt catcagtata 4861 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga 4921 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac 4981 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc 5041 ttctcttatg aagatccctc gacctgcagc ccaagcttgg atccctcgac agtactaagc 5101 tttactaggg acaggattgg tgacagaaaa gccccatcct taggcctcct ccttcctagt 5161 ctcctgatat tgggtctaac ccccacctcc tgttaggcag attccttatc tggtgacaca 5221 cccccatttc ctggagccat ctctctcctt gccagaacct ctaaggtttg cttacgatgg 5281 agccagagag gatcctggga gggagagctt ggcagggggt gggagggaag ggggggatgc 5341 gtgacctgcc cggttctcag tggccaccct gcgctaccct ctcccagaac ctgagctgct 5401 ctgacgcggc tgtctggtgc gtttcactga tcctggtgct gcagcttcct tacacttccc 5461 aagaggagaa gcagtttgga aaaacaaaat cagaataagt tggtcctgag ttctaacttt 5521 ggctcttcac ctttctagtc cccaatttat attgttcctc cgtgcgtcag ttttacctgt 5581 gagataaggc cagtagccag ccccgtcctg gcagggctgt ggtgaggagg ggggtgtccg 5641 tgtggaaaac tccctttgtg agaatggtgc gtcctaggtg ttcaccaggt cgtggccgcc 5701 tctactccct ttctctttct ccatccttct ttccttaaag agtccccagt gctatctggg 5761 acatattcct ccgcccagag cagggtcccg cttccctaag gccctgctct gggcttctgg 5821 gtttgagtcc ttggcaagcc caggagaggc gctcaggctt ccctgtcccc cttcctcgtc 5881 caccatctca tgcccctggc tctcctgccc cttccctaca ggggttcctg gctctgctct 5941 aagggcaagg gcgaattcgc ggccgctaaa ttcaattcgc cctatagtga gtcgtattac 6001 aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt 6061 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc 6121 gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt ttacacctat 6181 aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccg 6241 gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt 6301 gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg 6361 gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat 6421 gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg catgagatta 6481 tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca gaaacggtgc 6541 tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc aagcgcaaag 6601 agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc ggttttatgg 6661 acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg gaagccctgc 6721 aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg atcaagctct 6781 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt 6841 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc 6901 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag 6961 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg 7021 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac 7081 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc 7141 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc 7201 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc 7261 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg 7321 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat 7381 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc 7441 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa 7501 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat 7561 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattat taacgcttac 7621 aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatcagg 7681 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc 7741 aaatatgtat ccgctcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta 7801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca 7861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc 7921 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc 7981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc 8041 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat 8101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt 8161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc 8221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag 8281 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt 8341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac 8401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg 8461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat 8521 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc 8581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc 8641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa 8701 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg 8761 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga 8821 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac 8881 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt 8941 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc 9001 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat 9061 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag 9121 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc 9181 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag 9241 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac 9301 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg 9361 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct 9421 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc 9481 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga 9541 gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga 9601 agcggaag //
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