质粒名称: | pST1374-Lb-Cpf1-NLS |
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出品公司: | Addgene |
目录编号: | 107680 |
存储实验室: | Jia Chen |
载体骨架基本信息 |
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载体名称: | Addgene 44758 |
完整质粒大小: | 8768 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | LbCpf1 |
物种来源: | Lachnospiraceae bacterium |
插入大小: | 3711 bp |
启动子: | CMV |
克隆方法: | 限制性内切酶 |
5’克隆位点: | 未知 |
3’克隆位点: | 未知 |
5’测序引物 | CMV |
3’测序引物 | BGH poly reverse |
文献引用方法 |
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材料和方法部分: | pST1374-Lb-Cpf1-NLS was a gift from Jia Chen (Addgene plasmid # 107680 ; http://n2t.net/addgene:107680 ; RRID:Addgene_107680) |
参考文献部分: | Base editing with a Cpf1-cytidine deaminase fusion. Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, Huang X, Yang L, Chen J. Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102. 10.1038/nbt.4102 PubMed 29553573 |
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载体基本信息
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载体序列
LOCUS pST1374-Lb-Cpf1-NLS 8768 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses LbCpf1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pST1374-Lb-Cpf1-NLS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8768) AUTHORS Li X, Wang Y, Liu Y, Yang B, Wang X, Wei J, Lu Z, Zhang Y, Wu J, Huang X, Yang L, Chen J TITLE Base editing with a Cpf1-cytidine deaminase fusion. JOURNAL Nat Biotechnol. 2018 Mar 19. pii: nbt.4102. doi: 10.1038/nbt.4102. PUBMED 29553573 REFERENCE 2 (bases 1 to 8768) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8768 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(44..63) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 235..614 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 615..818 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 769..789 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 863..882 /label=T7 /note="T7 promoter, forward primer" promoter 863..881 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 907..4590 /codon_start=1 /product="CRISPR-associated protein from Lachnospiraceae bacterium ND2006 (Zetsche et al., 2015)" /label=LbCpf1 /translation="MSKLEKFTNCYSLSKTLRFKAIPVGKTQENIDNKRLLVEDEKRAE DYKGVKKLLDRYYLSFINDVLHSIKLKNLNNYISLFRKKTRTEKENKELENLEINLRKE IAKAFKGNEGYKSLFKKDIIETILPEFLDDKDEIALVNSFNGFTTAFTGFFDNRENMFS EEAKSTSIAFRCINENLTRYISNMDIFEKVDAIFDKHEVQEIKEKILNSDYDVEDFFEG EFFNFVLTQEGIDVYNAIIGGFVTESGEKIKGLNEYINLYNQKTKQKLPKFKPLYKQVL SDRESLSFYGEGYTSDEEVLEVFRNTLNKNSEIFSSIKKLEKLFKNFDEYSSAGIFVKN GPAISTISKDIFGEWNVIRDKWNAEYDDIHLKKKAVVTEKYEDDRRKSFKKIGSFSLEQ LQEYADADLSVVEKLKEIIIQKVDEIYKVYGSSEKLFDADFVLEKSLKKNDAVVAIMKD LLDSVKSFENYIKAFFGEGKETNRDESFYGDFVLAYDILLKVDHIYDAIRNYVTQKPYS KDKFKLYFQNPQFMGGWDKDKETDYRATILRYGSKYYLAIMDKKYAKCLQKIDKDDVNG NYEKINYKLLPGPNKMLPKVFFSKKWMAYYNPSEDIQKIYKNGTFKKGDMFNLNDCHKL IDFFKDSISRYPKWSNAYDFNFSETEKYKDIAGFYREVEEQGYKVSFESASKKEVDKLV EEGKLYMFQIYNKDFSDKSHGTPNLHTMYFKLLFDENNHGQIRLSGGAELFMRRASLKK EELVVHPANSPIANKNPDNPKKTTTLSYDVYKDKRFSEDQYELHIPIAINKCPKNIFKI NTEVRVLLKHDDNPYVIGIDRGERNLLYIVVVDGKGNIVEQYSLNEIINNFNGIRIKTD YHSLLDKKEKERFEARQNWTSIENIKELKAGYISQVVHKICELVEKYDAVIALEDLNSG FKNSRVKVEKQVYQKFEKMLIDKLNYMVDKKSNPCATGGALKGYQITNKFESFKSMSTQ NGFIFYIPAWLTSKIDPSTGFVNLLKTKYTSIADSKKFISSFDRIMYVPEEDLFEFALD YKNFSRTDADYIKKWKLYSYGNRIRIFRNPKKNNVFDWEEVCLTSAYKELFNKYGINYQ QGDIRALLCEQSDKAFYSSFMALMSLMLQMRNSITGRTDVDFLISPVKNSDGIFYDSRN YEAQENAILPKNADANGAYNIARKVLWAIGQFKKAEDEKLDKVKIAISNKEWLEYAQTS VKH" CDS 4597..4617 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4633..4674 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 4684..4701 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(4724..4741) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4730..4954 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5000..5428 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5087..5106) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5297..5318 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5437..5457) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5442..5771 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 5622..5757 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5684..5703 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 5819..5866 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 5885..6283 /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" polyA_signal 6441..6562 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6478..6497) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6532..6551 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6611..6627) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6611..6627) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6624..6646) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6635..6651 /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(6659..6689) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6704..6725 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6842..6859) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7013..7601) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7093..7112) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7772..8632) /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 8395..8414 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8633..8737) /gene="bla" /label=AmpR promoter ORIGIN 1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg 61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg 121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc 181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt 241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc 361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc 421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt 481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt 541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca 601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg 661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc 721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg 781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca 841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc 901 gccaccatgt caaagctgga gaaattcacc aactgttata gcctgtctaa gaccctgcgc 961 ttcaaggcaa tcccagtggg caagacacaa gagaacattg acaacaaacg gctcctggtg 1021 gaggatgaga agagggctga agattacaag ggcgttaaga agctgctgga taggtactat 1081 ctgtcattca tcaacgatgt cctccacagt atcaagctga agaatctgaa caattacatt 1141 tctctgttcc ggaagaagac acggaccgag aaggagaaca aagagctgga gaatctggag 1201 atcaacctga ggaaagaaat agctaaggct ttcaaaggga acgagggtta caagtccctg 1261 ttcaagaaag acattatcga gactattctg cctgagttcc tggacgataa agatgagatc 1321 gccctcgtca attccttcaa tgggtttacc acagccttta ccggcttctt cgacaataga 1381 gagaatatgt tctctgaaga ggccaaatcc actagcatcg cctttcgctg cataaacgag 1441 aacctgacta ggtacatcag caatatggac atctttgaga aagtcgatgc catattcgac 1501 aaacatgagg tgcaggagat taaggagaag atcctgaact cagattacga tgtcgaagat 1561 ttcttcgagg gagagttctt caacttcgtg ctcacacaag agggcattga tgtgtacaat 1621 gcaatcattg gagggttcgt gacagagagt ggcgagaaga taaagggcct gaacgagtat 1681 atcaacctct acaaccagaa aaccaagcag aaactgccta agttcaagcc actgtacaaa 1741 caagtgctct cagataggga aagcctgagc ttctacggtg aagggtatac atcagatgaa 1801 gaagtgctcg aagtgttccg caacaccctc aataagaaca gtgaaatctt ctcttcaatc 1861 aagaagctgg agaaactgtt caagaatttc gatgagtact cctctgccgg aatctttgtg 1921 aagaatggcc ctgcaatatc cactattagc aaagacatct ttggcgagtg gaacgttatc 1981 agggataagt ggaatgccga gtacgatgat attcatctca agaagaaagc cgtggttaca 2041 gagaaatacg aggatgatag acgcaagagc tttaagaaga ttggtagctt ctctctcgaa 2101 cagctgcagg agtacgccga cgctgacctg tcagtcgtgg agaaactcaa ggagatcata 2161 atccagaagg tggatgaaat ctacaaagtg tatggaagct ctgagaaact cttcgatgca 2221 gactttgttc tggagaagag tctgaagaag aacgacgcag tggttgctat catgaaggac 2281 ctgctggatt ctgttaagtc tttcgagaat tacattaagg cattctttgg tgaagggaag 2341 gagacaaata gggacgagag cttctatggc gactttgttc tggcctacga catcctcctc 2401 aaggttgacc acatctatga cgctatacgg aattacgtta cccagaagcc ctatagcaaa 2461 gacaagttca agctgtattt ccagaatcca cagtttatgg gtgggtggga taaagacaaa 2521 gaaacagatt acagggccac tatcctgcgg tacggcagca aatactatct ggctatcatg 2581 gataagaagt acgccaaatg cctccagaag atcgacaagg acgacgtgaa cggtaactac 2641 gagaagatca attacaagct cctgccagga cctaacaaga tgctgcccaa ggtgttcttc 2701 tccaagaaat ggatggccta ctataaccca agcgaggaca ttcagaagat atacaagaat 2761 gggacattca agaagggcga tatgttcaac ctcaacgact gccacaagct gattgatttc 2821 ttcaaggata gcatttctcg ctatcccaag tggtctaatg catacgattt caacttcagc 2881 gagactgaga agtacaaaga catcgctggc ttctaccggg aggtggaaga gcaaggctat 2941 aaggtgtcat tcgaatccgc ttctaagaag gaagtggata agctcgtgga agagggtaag 3001 ctgtacatgt tccagatata caacaaagac ttcagcgata agagccacgg cactccaaac 3061 ctccatacta tgtatttcaa gctgctgttt gacgagaaca accacggaca gattaggctg 3121 tcaggaggcg cagaactctt catgcgcaga gcttcactga agaaggagga actcgttgtc 3181 cacccagcca atagccctat agccaataag aatccagaca atcctaagaa aaccactact 3241 ctgtcttacg atgtgtataa ggataagaga ttctctgaag atcagtacga actgcacata 3301 cccattgcca ttaacaagtg ccctaagaac atcttcaaga ttaacacaga ggttagagtg 3361 ctcctgaaac acgacgataa cccttatgtt ataggcattg atcgcggaga gagaaacctg 3421 ctgtacatcg tcgtggtgga cggcaaaggc aacatcgtgg aacagtacag tctcaatgaa 3481 atcattaaca atttcaacgg aatccgcatt aagaccgact accattctct cctcgacaag 3541 aaggagaaag aaaggttcga agcaagacag aattggacaa gtatagagaa tatcaaagaa 3601 ctgaaggctg ggtacatctc tcaggttgtg cacaagatat gtgagctggt ggagaagtac 3661 gacgctgtta tcgccctcga ggacctgaat agcggcttca agaactccag ggtgaaggtg 3721 gagaagcagg tgtatcagaa gttcgagaag atgctgatcg acaagctcaa ctatatggtg 3781 gacaagaaat ccaatccttg cgctactggt ggagccctga agggctatca aatcaccaat 3841 aagttcgaat ctttcaagtc tatgagcacc cagaatggct tcatcttcta catacccgca 3901 tggctgacat ccaagattga tccctctacc ggatttgtta atctgctcaa gactaagtac 3961 acctctattg ctgactcaaa gaagttcata tcatcatttg accgcatcat gtacgtgcca 4021 gaagaggacc tgttcgagtt tgccctggat tacaagaatt tctctcggac tgacgccgac 4081 tacatcaaga agtggaagct ctactcttat ggtaatcgga ttcgcatatt ccgcaatccc 4141 aagaagaata acgtgttcga ttgggaggaa gtttgcctca ccagcgctta caaggagctg 4201 ttcaataagt atgggattaa ctaccagcag ggcgacataa gagccctgct gtgcgaacaa 4261 tctgataagg cattctattc ctctttcatg gcactgatgt cactgatgct gcaaatgcgc 4321 aattccatca ccggaagaac agacgtggac tttctgatct ctcctgtcaa gaactcagat 4381 ggcatcttct acgattcccg caactatgaa gcacaggaga atgctatcct gcctaagaat 4441 gccgatgcaa atggagccta taacatcgcc agaaaggtcc tctgggccat aggacaattc 4501 aagaaagctg aagatgagaa gctggacaag gtgaagatcg ccatttcaaa caaagagtgg 4561 ctcgaatatg ctcagacctc agtgaagcat ggatcaccca agaagaaacg gaaagtgtga 4621 gggcccttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc 4681 ggtcatcatc accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct 4741 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc 4801 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt 4861 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat 4921 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg 4981 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg 5041 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc 5101 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc 5161 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt 5221 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag 5281 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg 5341 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag 5401 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg 5461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag 5521 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 5581 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc 5641 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg 5701 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag 5761 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt 5821 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta 5881 aaccatggcc aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac 5941 aatcaacagc atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga 6001 cggccgcatc ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact 6061 cgtggtgctg ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat 6121 cggaaatgag aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga 6181 tctgcatcct gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg 6241 gattcgtgaa ttgctgccct ctggttatgt gtgggagggc taagcacttc gtggccgagg 6301 agcaggactg acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg 6361 gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc 6421 tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca 6481 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt 6541 ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg 6601 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca 6661 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca 6721 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc 6781 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt 6841 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact 6901 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag 6961 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata 7021 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc 7081 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg 7141 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc 7201 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg 7261 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc 7321 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga 7381 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg 7441 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa 7501 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg 7561 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt 7621 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat 7681 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct 7741 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta 7801 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa 7861 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac 7921 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa 7981 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag 8041 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg 8101 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag 8161 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg 8221 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc 8281 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat 8341 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata 8401 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa 8461 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca 8521 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc 8581 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc 8641 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg 8701 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac 8761 ctgacgtc //
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