质粒名称: | pcDNA-dCas9 |
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出品公司: | Addgene |
目录编号: | 47106 |
存储实验室: | Charles Gersbach |
载体骨架基本信息 |
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载体名称: | pcDNA3.1 |
载体出品公司: | Invitrogen |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | 9575 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达,CRISPR |
筛选标记: | -- |
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: | dCas9 |
物种来源: | S. Pyogenes |
插入大小: | 4236 bp |
基因突变: | D10A, H840A (catalytically inactive) |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: |
标志(插入时N端子)
SV40 NLS(N端子插入)
SV40 NLS(C端子插入)
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克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | CMV-F |
3' 测序引物 | BGH-rev |
文献引用方法 |
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材料和方法部分: | pcDNA-dCas9 was a gift from Charles Gersbach (Addgene plasmid # 47106 ; http://n2t.net/addgene:47106 ; RRID:Addgene_47106) |
参考文献部分: | RNA-guided gene activation by CRISPR-Cas9-based transcription factors. Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein LR, Thakore PI, Glass KA, Ousterout DG, Leong KW, Guilak F, Crawford GE, Reddy TE, Gersbach CA. Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600. 10.1038/nmeth.2600 PubMed 23892895 |
买家导航
pcDNAdCas9载体基本信息
pcDNAdCas9载体图谱质粒图谱和多克隆位点信息
pcDNAdCas9载体简介
pcDNAdCas9载体序列
LOCUS pcDNA-dCas9 9575 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses inactivated S. pyogenes dCas9 (D10A, H840A) in mammalian cells. ACCESSION . VERSION . KEYWORDS pcDNA-dCas9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9575) AUTHORS Perez-Pinera P, Kocak DD, Vockley CM, Adler AF, Kabadi AM, Polstein LR, Thakore PI, Glass KA, Ousterout DG, Leong KW, Guilak F, Crawford GE, Reddy TE, Gersbach CA TITLE RNA-guided gene activation by CRISPR-Cas9-based transcription factors. JOURNAL Nat Methods. 2013 Jul 25. doi: 10.1038/nmeth.2600. PUBMED 23892895 REFERENCE 2 (bases 1 to 9575) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9575 /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..972 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 979..999 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 1009..5112 /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="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGD" CDS 5113..5133 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" primer_bind complement(5169..5186) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5175..5399 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 5445..5873 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5532..5551) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5742..5763 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5882..5902) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 5887..6216 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6067..6202 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6129..6148 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 6283..7077 /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 GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(6337..6356) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6947..6966 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 7251..7372 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(7288..7307) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7342..7361 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7421..7437) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7421..7437) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7434..7456) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7445..7461 /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(7469..7499) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7514..7535 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7652..7669) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7823..8408) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7903..7922) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8579..9439) /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 9202..9221 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9440..9544) /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 gccatggact acaaagacca tgacggtgat tataaagatc atgacatcga ttacaaggat 961 gacgatgaca agatggcccc caagaagaag aggaaggtgg gccgcggaat ggataagaaa 1021 tactcaatag gcttagctat cggcacaaat agcgtcggat gggcggtgat cactgatgaa 1081 tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca cagtatcaaa 1141 aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc gactcgtctc 1201 aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta tctacaggag 1261 attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact tgaagagtct 1321 tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa tatagtagat 1381 gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa attggtagat 1441 tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat gattaagttt 1501 cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt ggacaaacta 1561 tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat taacgcaagt 1621 ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg attagaaaat 1681 ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct cattgctttg 1741 tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga tgctaaatta 1801 cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca aattggagat 1861 caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt actttcagat 1921 atcctaagag taaatactga aataactaag gctcccctat cagcttcaat gattaaacgc 1981 tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca acaacttcca 2041 gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg ttatattgat 2101 gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga aaaaatggat 2161 ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa gcaacggacc 2221 tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc tattttgaga 2281 agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga aaaaatcttg 2341 acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg ttttgcatgg 2401 atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt tgtcgataaa 2461 ggtgcttcag ctcaatcatt tattgaacgc atgacaaact ttgataaaaa tcttccaaat 2521 gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta taacgaattg 2581 acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc aggtgaacag 2641 aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt taagcaatta 2701 aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc aggagttgaa 2761 gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat taaagataaa 2821 gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt aacattgacc 2881 ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca cctctttgat 2941 gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggacg tttgtctcga 3001 aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga ttttttgaaa 3061 tcagatggtt ttgccaatcg caattttatg cagctgatcc atgatgatag tttgacattt 3121 aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca tgaacatatt 3181 gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt aaaagttgtt 3241 gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat tgaaatggca 3301 cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat gaaacgaatc 3361 gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt tgaaaatact 3421 caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga catgtatgtg 3481 gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatgccat tgttccacaa 3541 agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga taaaaatcgt 3601 ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa ctattggaga 3661 caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac gaaagctgaa 3721 cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt ggttgaaact 3781 cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac taaatacgat 3841 gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa attagtttct 3901 gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta ccatcatgcc 3961 catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata tccaaaactt 4021 gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat gattgctaag 4081 tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa tatcatgaac 4141 ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc tctaatcgaa 4201 actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc cacagtgcgc 4261 aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca gacaggcgga 4321 ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc tcgtaaaaaa 4381 gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta ttcagtccta 4441 gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa agagttacta 4501 gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt tttagaagct 4561 aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata tagtcttttt 4621 gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca aaaaggaaat 4681 gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca ttatgaaaag 4741 ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca gcataagcat 4801 tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat tttagcagat 4861 gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc aatacgtgaa 4921 caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc cgctgctttt 4981 aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga agttttagat 5041 gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga tttgagtcag 5101 ctaggaggtg accccaagaa gaagaggaag gtgggctgat ctagacttaa gtttaaaccg 5161 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 5221 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat 5281 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 5341 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 5401 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg 5461 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc 5521 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc 5581 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct 5641 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac 5701 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac 5761 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat 5821 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg 5881 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg 5941 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca 6001 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact 6061 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta 6121 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag 6181 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc 6241 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga 6301 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa 6361 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt 6421 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg 6481 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa 6541 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac 6601 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt 6661 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact 6721 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg 6781 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg 6841 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc 6901 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt 6961 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc 7021 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg 7081 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg 7141 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct 7201 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta 7261 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 7321 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 7381 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt 7441 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag 7501 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt 7561 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag 7621 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg 7681 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat 7741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta 7801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa 7861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc 7921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt 7981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca 8041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg 8101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat 8161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta 8221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct 8281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac 8341 aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 8401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 8461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 8521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 8581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 8641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 8701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 8761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 8821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 8881 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 8941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 9001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 9061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 9121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 9181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 9241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 9301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 9361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 9421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 9481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 9541 cgcgcacatt tccccgaaaa gtgccacctg acgtc //
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pcDNAdCas9载体质粒应用举例
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