质粒名称: | pHAGE-TO-dCas9 |
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出品公司: | 爱迪基因 |
目录编号: | 75381 |
存储实验室: | Thoru Pederson |
载体骨架基本信息 |
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载体名称: | pHAGE-DEST |
载体抗性: | 氨苄霉素 |
空载体大小: | 6000 bp |
完整质粒大小: | 10829 bp |
载体类型: | 哺乳细胞表达,Lentiviral |
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: | Sp dCas9 |
物种来源: | Synthetic |
插入大小: | 4101 bp |
启动子 | CMV-TO |
标签/融合蛋白: | no |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | EcoRI(未破坏) |
3' 克隆位点: | XbaI(未破坏) |
5' 测序引物 | cgaaggaatagaagaagaaggtgga |
3' 测序引物 | CCAAAGGGAGATCCGACTCG |
文献引用方法 |
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材料和方法部分: | pHAGE-TO-dCas9 was a gift from Thoru Pederson (爱迪基因 plasmid # 75381) |
参考文献部分: | Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T. Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. 10.1038/nbt.3526 PubMed 27088723 |
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 pHAGE-TO-dCas9 10835 bp ds-DNA circular SYN 10-JAN-2018 DEFINITION dCas9. ACCESSION . VERSION . KEYWORDS pHAGE-TO-dCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10835) AUTHORS Ma H, Tu LC, Naseri A, Huisman M, Zhang S, Grunwald D, Pederson T TITLE Multiplexed labeling of genomic loci with dCas9 and engineered sgRNAs using CRISPRainbow. JOURNAL Nat Biotechnol. 2016 Apr 18. doi: 10.1038/nbt.3526. PUBMED 27088723 COMMENT This file is created by Vector NTI http://www.biofeng.com/ COMMENT ORIGDB|GenBank COMMENT VNTAUTHORNAME|biofeng.com| COMMENT VNTNAME|pHAGE-TO-dCas9| FEATURES Location/Qualifiers source 1..10835 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." misc_feature 2058..2175 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" enhancer 2299..2602 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2603..2806 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2757..2777 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" protein_bind 2808..2826 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 2829..2847 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" primer_bind 2857..2881 /label=LNCX /note="Human CMV promoter, forward primer" regulatory 2974..2983 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2986..3006 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 3025..7125 /codon_start=1 /product="catalytically dead mutant of the Cas9 endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=dCas9 /note="RNA-guided DNA-binding protein that lacks endonuclease activity due to the D10A mutation in the RuvC catalytic domain and the H840A mutation in the HNH catalytic domain" /translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 7144..7164 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" misc_feature 7225..7813 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(7278..7298) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(7696..7707) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 7888..8121 /label=3' LTR (Delta-U3) /note="self-inactivating 3' long terminal repeat (LTR) from HIV-1" primer_bind complement(8240..8258) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 8326..8430 /gene="bla" /label=AmpR promoter CDS 8431..9291 /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind complement(8649..8668) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9462..10050 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9951..9970 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10204..10221 /label=L4440 /note="L4440 vector, forward primer" primer_bind 10313..10332 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" protein_bind 10461..10482 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 10497..10527 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 10535..10551 /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 10540..10562 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 10559..10575 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 10559..10575 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind 10625..10644 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccgaa ttgacaaatg 2041 gcagtattca tccacaattt taaaagaaaa ggggggattg gggggtacag tgcaggggaa 2101 agaatagtag acataatagc aacagacata caaactaaag aattacaaaa acaaattaca 2161 aaaattcaaa attttcgggt ttattacagg gacagcagag atccagtttg gactagcgat 2221 gctagctgat gcgggcccta gttattaata gtaatcaatt acggggtcat tagttcatag 2281 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc 2341 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg 2401 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca 2461 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc 2521 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt 2581 attagtcatc gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata 2641 gcggtttgac tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt 2701 ttggcaccaa aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca 2761 aatgggcggt aggcgtgtac ggtgggaggt ctatataagc agagctctcc ctatcagtga 2821 tagagatctc cctatcagtg atagagatcg tcgacgagct cgtttagtga accgtcagat 2881 cgcctggaga cgccatccac gctgttttga cctccataga agacaccggg accgatccag 2941 cctaccggtt aaggtgaacg cgtgcaggct ggcgccacca tggcccctaa aaagaaacgc 3001 aaggtcgaat tcggaagcgg aagcgacaag aagtactcca ttgggctcgc tatcggtacc 3061 aacagcgtcg gctgggccgt cattacggac gagtacaagg tgccgagcaa aaaattcaaa 3121 gttctgggca ataccgatcg ccacagcata aagaagaacc tcattggagc cctcctgttc 3181 gactccgggg agacggccga agccacgcgg ctcaaaagaa cagcacggcg cagatatacc 3241 cgcagaaaga atcggatctg ctacctgcag gagatcttta gtaatgagat ggctaaggtg 3301 gatgactctt tcttccatag gctggaggag tcctttttgg tggaggagga taaaaagcac 3361 gagcgccacc caatctttgg caatatcgtg gacgaggtgg cgtaccatga aaagtaccca 3421 accatatatc atctgaggaa gaagctggta gacagtactg ataaggctga cttgcggttg 3481 atctatctcg cgctggcgca catgatcaaa tttcggggac acttcctcat cgagggggac 3541 ctgaacccag acaacagcga tgtcgacaaa ctctttatcc aactggttca gacttacaat 3601 cagcttttcg aggagaaccc gatcaacgca tccggcgttg acgccaaagc aatcctgagc 3661 gctaggctgt ccaaatcccg gcggctcgaa aacctcatcg cacagctccc tggggagaag 3721 aagaacggcc tgtttggtaa tcttatcgcc ctgtcactcg ggctgacccc caactttaaa 3781 tctaacttcg acctggccga agatgccaag ctgcaactga gcaaagacac ctacgatgat 3841 gatctcgaca atctgctggc ccagatcggc gaccagtacg cagacctttt tttggcggca 3901 aagaacctgt cagacgccat tctgctgagt gatattctgc gagtgaacac ggagatcacc 3961 aaagctccgc tgagcgctag tatgatcaag cgctatgatg agcaccacca agacttgact 4021 ttgctgaagg cccttgtcag acagcaactg cctgagaagt acaaggaaat tttcttcgat 4081 cagtctaaaa atggctacgc cggatacatt gacggcggag caagccagga ggaattttac 4141 aaatttatta agcccatctt ggaaaaaatg gacggcaccg aggagctgct ggtaaagctg 4201 aacagagaag atctgttgcg caaacagcgc actttcgaca atggaagcat cccccaccag 4261 attcacctgg gcgaactgca cgctatcctc aggcggcaag aggatttcta cccctttttg 4321 aaagataaca gggaaaagat tgagaaaatc ctcacatttc ggatacccta ctatgtaggc 4381 cccctcgctc ggggaaattc cagattcgcg tggatgactc gcaaatcaga agagaccatc 4441 actccctgga acttcgagga agtcgtggat aagggggcct ctgcccagtc cttcatcgaa 4501 aggatgacta actttgataa aaatctgcct aacgaaaagg tgcttcctaa acactctctg 4561 ctgtacgagt acttcacagt ttataacgag ctcaccaagg tcaaatacgt cacagaaggg 4621 atgagaaagc cagcattcct gtctggagag cagaagaaag ctatcgtgga cctcctcttc 4681 aagacgaacc ggaaagttac cgtgaaacag ctcaaagaag actatttcaa aaagattgaa 4741 tgtttcgact ctgttgaaat cagcggagtg gaggatcgct tcaacgcatc cctgggaacg 4801 tatcacgatc tcctgaaaat cattaaagac aaggacttcc tggacaatga ggagaacgag 4861 gacattcttg aggacattgt cctcaccctt acgttgtttg aagataggga gatgattgaa 4921 gaacgcttga aaacttacgc tcatctcttc gacgacaaag tcatgaaaca gctcaagaga 4981 cgccgatata caggatgggg gcggctgtca agaaaactga tcaatggcat ccgagacaag 5041 cagagtggaa agacaatcct ggattttctt aagtccgatg gatttgccaa ccggaacttc 5101 atgcagttga tccatgatga ctctctcacc tttaaggagg acatccagaa agcacaagtt 5161 tctggccagg gggacagtct tcacgagcac atcgctaatc ttgcaggtag cccagctatc 5221 aaaaagggaa tactgcagac cgttaaggtc gtggatgaac tcgtcaaagt aatgggaagg 5281 cataagcccg agaatatcgt tatcgagatg gcccgagaga accaaactac ccagaaggga 5341 cagaagaaca gtagggaaag gatgaagagg attgaagagg gtataaaaga actggggtcc 5401 caaatcctta aggaacaccc agttgaaaac acccagcttc agaatgagaa gctctacctg 5461 tactacctgc agaacggcag ggacatgtac gtggatcagg aactggacat caaccggttg 5521 tccgactacg acgtggatgc tatcgtgccc caaagctttc tcaaagatga ttctattgat 5581 aataaagtgt tgacaagatc cgataaaaat agagggaaga gtgataacgt cccctcagaa 5641 gaagttgtca agaaaatgaa aaattattgg cggcagctgc tgaacgccaa actgatcaca 5701 caacggaagt tcgataatct gactaaggct gaacgaggtg gcctgtctga gttggataaa 5761 gccggcttca tcaaaaggca gcttgttgag acacgccaga tcaccaagca cgtggcccaa 5821 attctcgatt cacgcatgaa caccaagtac gatgaaaatg acaaactgat tcgagaggtg 5881 aaagttatta ctctgaagtc taagctggtc tcagatttca gaaaggactt tcagttttat 5941 aaggtgagag agatcaacaa ttaccaccat gcgcatgatg cctacctgaa tgcagtggta 6001 ggcactgcac ttatcaaaaa atatcccaag ctggaatctg aatttgttta cggagactat 6061 aaagtgtacg atgttaggaa aatgatcgca aagtctgagc aggaaatagg caaggccacc 6121 gctaagtact tcttttacag caatattatg aattttttca agaccgagat tacactggcc 6181 aatggagaga ttcggaagcg accacttatc gaaacaaacg gagaaacagg agaaatcgtg 6241 tgggacaagg gtagggattt cgcgacagtc cgcaaggtcc tgtccatgcc gcaggtgaac 6301 atcgttaaaa agaccgaagt acagaccgga ggcttctcca aggaaagtat cctcccgaaa 6361 aggaacagcg acaagctgat cgcacgcaaa aaagattggg accccaagaa atacggcgga 6421 ttcgattctc ctacagtcgc ttacagtgta ctggttgtgg ccaaagtgga gaaagggaag 6481 tctaaaaaac tcaaaagcgt caaggaactg ctgggcatca caatcatgga gcgatccagc 6541 ttcgagaaaa accccatcga ctttctcgaa gcgaaaggat ataaagaggt caaaaaagac 6601 ctcatcatta agctgcccaa gtactctctc tttgagcttg aaaacggccg gaaacgaatg 6661 ctcgctagtg cgggcgagct gcagaaaggt aacgagctgg cactgccctc taaatacgtt 6721 aatttcttgt atctggccag ccactatgaa aagctcaaag ggtctcccga agataatgag 6781 cagaagcagc tgttcgtgga acaacacaaa cactaccttg atgagatcat cgagcaaata 6841 agcgagttct ccaaaagagt gatcctcgcc gacgctaacc tcgataaggt gctttctgct 6901 tacaataagc acagggataa gcccatcagg gagcaggcag aaaacattat ccacttgttt 6961 actctgacca acttgggcgc gcctgcagcc ttcaagtact tcgacaccac catagacaga 7021 aagcggtaca cctctacaaa ggaggtcctg gacgccacac tgattcatca gtcaattacg 7081 gggctctatg aaacaagaat cgacctctct cagctcggtg gagacggctc tactagtggc 7141 tctcccaaga agaagaggaa ggtaggatcc ggatctggcg gccgctctgg catcgatggg 7201 agcctcgagt agggtaatct agataatcaa cctctggatt acaaaatttg tgaaagattg 7261 actggtattc ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct 7321 ttgtatcatg ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg 7381 ttgctgtctc tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact 7441 gtgtttgctg acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc 7501 gggactttcg ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc 7561 cgctgctgga caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa 7621 tcatcgtcct ttccttggct gctcgcctgt gttgccacct ggattctgcg cgggacgtcc 7681 ttctgctacg tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg 7741 gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg 7801 gccgcctccc cgcctgagat cctttaagac caatgactta caaggcagct gtagatctta 7861 gccacttttt aaaagaaaag gggggactgg aagggctaat tcactcccaa cgaagacaag 7921 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct 7981 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag 8041 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt 8101 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac 8161 ttgcaaagaa atgaatatca gagagtgaga ggcccgggtt aattaaggaa agggctagat 8221 cattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata 8281 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt 8341 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa 8401 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt 8461 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa 8521 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac 8581 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt 8641 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt 8701 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat 8761 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac 8821 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg 8881 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc 8941 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa 9001 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag 9061 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct 9121 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat 9181 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa 9241 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac 9301 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc 9361 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc 9421 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg 9481 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg 9541 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca 9601 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg 9661 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg 9721 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga 9781 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac 9841 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat 9901 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc 9961 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga 10021 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc 10081 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg 10141 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag 10201 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc 10261 gcgcgttggc cgattcatta atgcagcaag ctcatggctg actaattttt tttatttatg 10321 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg 10381 gaggcctagg cttttgcaaa aagctccccg tggcacgaca ggtttcccga ctggaaagcg 10441 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta 10501 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca 10561 ggaaacagct atgacatgat tacgaatttc acaaataaag catttttttc actgcattct 10621 agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa ctggataact 10681 caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac tgccaattac 10741 ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt ttaaagaaat 10801 tgtatttgtt aaatatgtac tacaaactta gtagt //
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