质粒名称: | pPlatTET-gRNA2 |
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出品公司: | 爱迪基因 |
目录编号: | 82559 |
存储实验室: | Izuho Hatada |
载体骨架基本信息 |
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载体名称: | pCAG |
载体出品公司: | -- |
载体抗性: | 卡那霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
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: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | dCas9-5xPlat2AflD-P2A-scFvGCN4sfGFPTET1CD |
物种来源: | -- |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pPlatTET-gRNA2 was a gift from Izuho Hatada (Addgene plasmid # 82559 ; http://n2t.net/addgene:82559 ; RRID:Addgene_82559) |
参考文献部分: | Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions. Morita S, Noguchi H, Horii T, Nakabayashi K, Kimura M, Okamura K, Sakai A, Nakashima H, Hata K, Nakashima K, Hatada I. Nat Biotechnol. 2016 Aug 29. doi: 10.1038/nbt.3658. 10.1038/nbt.3658 PubMed 27571369 |
买家导航
pPlatTETgRNA2载体基本信息
pPlatTETgRNA2载体图谱质粒图谱和多克隆位点信息
pPlatTETgRNA2载体简介
pPlatTETgRNA2载体序列
LOCUS pPlatTET-gRNA2 14290 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION All in one vector which contains Cas9 peptide array (linker length: 22aa), antibody-sfGFP-TET1CD, and gRNA expression system.. ACCESSION . VERSION . KEYWORDS pPlatTET-gRNA2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 14290) AUTHORS Morita S, Noguchi H, Horii T, Nakabayashi K, Kimura M, Okamura K, Sakai A, Nakashima H, Hata K, Nakashima K, Hatada I TITLE Targeted DNA demethylation in vivo using dCas9-peptide repeat and scFv-TET1 catalytic domain fusions. JOURNAL Nat Biotechnol. 2016 Aug 29. doi: 10.1038/nbt.3658. PUBMED 27571369 REFERENCE 2 (bases 1 to 14290) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..14290 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 1..304 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 582..1598 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1522..1544 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1606..1625 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1674..1694 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" regulatory 1743..1752 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1749..5852 /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 5856..5882 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 5901..5921 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 5928..5948 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 6012..6068 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 6135..6191 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 6258..6314 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 6381..6437 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 6504..6560 /codon_start=1 /product="GCN4 peptide optimized to improve solubility while preserving binding to an scFv-GFP fusion protein (Tanenbaum et al., 2014)" /label=GCN4_v4 /translation="EELLSKNYHLENEVARLKK" CDS 6648..6704 /codon_start=1 /product="2A peptide from porcine teschovirus-1 polyprotein" /label=P2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="ATNFSLLKQAGDVEENPGP" CDS 7452..7478 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS 7569..8279 /codon_start=1 /product="GFP variant that folds robustly even when fused to poorly folded proteins (Pedelacq et al., 2006)" /label=superfolder GFP /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF VTAAGITHGMDELYK" primer_bind complement(7596..7624) /label=GFP-R /note="GFP, reverse primer. Does NOT anneal to EGFP" primer_bind 8219..8239 /label=GFP-F /note="GFP, forward primer. Does NOT anneal to EGFP" polyA_signal 10606..10687 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10643..10662) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10697..10716 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 10804..10824 /label=hU6-F /note="Human U6 promoter, forward primer" primer_bind 10975..10994 /label=LKO.1 5' /note="Human U6 promoter, forward primer" protein_bind complement(11101..11125) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" rep_origin complement(11146..11601) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(11283..11304) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 11495..11514 /label=F1ori-R /note="F1 origin, reverse primer" promoter 11628..11732 /gene="bla" /label=AmpR promoter promoter 11734..12091 /label=SV40 promoter /note="SV40 enhancer and early promoter" primer_bind complement(11757..11777) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" rep_origin 11942..12077 /label=SV40 ori /note="SV40 origin of replication" primer_bind 12004..12023 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 12126..12920 /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(12180..12199) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 12790..12809 /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind complement(13108..13127) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 13152..13199 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 13528..14116 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 14017..14036 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt 61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca 121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc 181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta 241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac 301 catggtcgag gtgagcccca cgttctgctt cactctcccc atctcccccc cctccccacc 361 cccaattttg tatttattta ttttttaatt attttgtgca gcgatggggg cggggggggg 421 gggggggcgc gcccggcggg ggggggcggg gcgaggggcg gggcggggcg aggcggagag 481 gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc 541 ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc gggagtcgct gcgcgctgcc 601 ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc tgactgaccg 661 cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt aattagcgct 721 tggtttaatg acggcttgtt tcttttctgt ggctgcgtga aagccttgag gggctccggg 781 agggcccttt gtgcgggggg agcggctcgg ggggtgcgtg cgtgtgtgtg tgcgtgggga 841 gcgccgcgtg cggctccgcg ctgcccggcg gctgtgagcg ctgcgggcgc ggcgcggggc 901 tttgtgcgct ccgcagtgtg cgcgagggga gcgcggccgg gggcggtgcc ccgcggtgcg 961 gggggggctg cgaggggaac aaaggctgcg tgcggggtgt gtgcgtgggg gggtgagcag 1021 ggggtgtggg cgcgtcggtc gggctgcaac cccccctgca cccccctccc cgagttgctg 1081 agcacggccc ggcttcgggt gcggggctcc gtacggggcg tggcgcgggg ctcgccgtgc 1141 cgggcggggg gtggcggcag gtgggggtgc cgggcggggc ggggccgcct cgggccgggg 1201 agggctcggg ggaggggcgc ggcggccccc ggagcgccgg cggctgtcga ggcgcggcga 1261 gccgcagcca ttgcctttta tggtaatcgt gcgagagggc gcagggactt cctttgtccc 1321 aaatctgtgc ggagccgaaa tctgggaggc gccgccgcac cccctctagc gggcgcgggg 1381 cgaagcggtg cggcgccggc aggaaggaaa tgggcgggga gggccttcgt gcgtcgccgc 1441 gccgccgtcc ccttctccct ctccagcctc ggggctgtcc gcggggggac ggctgccttc 1501 gggggggacg gggcagggcg gggttcggct tctggcgtgt gaccggcggc tctagagcct 1561 ctgctaacca tgttcatgcc ttcttctttt tcctacagct cctgggcaac gtgctggtta 1621 ttgtgctgtc tcatcatttt ggcaaagaat tcccccgcga tcgcgccacc atgcccaaga 1681 agaagcgcaa ggtgggacgc gtctgcagga tatcaagctt gcggtaccgc gggcccggga 1741 tcgccaccat ggacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct 1801 gggccgtgat caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca 1861 ccgaccggca cagcatcaag aagaacctga tcggcgccct gctgttcgac agcggagaaa 1921 cagccgaggc cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc 1981 ggatctgcta tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct 2041 tccacagact ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca 2101 tcttcggcaa catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc 2161 tgagaaagaa actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc 2221 tggcccacat gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca 2281 acagcgacgt ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg 2341 aaaaccccat caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca 2401 agagcagacg gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt 2461 tcggcaacct gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc 2521 tggccgagga tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc 2581 tgctggccca gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg 2641 acgccatcct gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga 2701 gcgcctctat gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc 2761 tcgtgcggca gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg 2821 gctacgccgg ctacatcgat ggcggagcca gccaggaaga gttctacaag ttcatcaagc 2881 ccatcctgga aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc 2941 tgctgcggaa gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag 3001 agctgcacgc cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg 3061 aaaagatcga gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg 3121 gaaacagcag attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact 3181 tcgaggaagt ggtggacaag ggcgccagcg cccagagctt catcgagcgg atgaccaact 3241 tcgataagaa cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact 3301 tcaccgtgta caacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg 3361 ccttcctgag cggcgagcag aaaaaagcca tcgtggacct gctgttcaag accaaccgga 3421 aagtgaccgt gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg 3481 tggaaatctc cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc 3541 tgaaaattat caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag 3601 atatcgtgct gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa 3661 cctatgccca cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg 3721 gctggggcag gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga 3781 caatcctgga tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc 3841 acgacgacag cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg 3901 atagcctgca cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc 3961 tgcagacagt gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga 4021 acatcgtgat cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc 4081 gcgagagaat gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag 4141 aacaccccgt ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga 4201 atgggcggga tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg 4261 tggacgctat cgtgcctcag agctttctga aggacgactc catcgataac aaagtgctga 4321 ctcggagcga caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga 4381 agatgaagaa ctactggcgc cagctgctga atgccaagct gattacccag aggaagttcg 4441 acaatctgac caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca 4501 agagacagct ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc 4561 ggatgaacac taagtacgac gagaacgaca aactgatccg ggaagtgaaa gtgatcaccc 4621 tgaagtccaa gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga 4681 tcaacaacta ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga 4741 tcaaaaagta ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg 4801 tgcggaagat gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct 4861 tctacagcaa catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc 4921 ggaagcggcc tctgatcgag acaaacggcg aaacaggcga gatcgtgtgg gataagggcc 4981 gggactttgc caccgtgcgg aaagtgctgt ctatgcccca agtgaatatc gtgaaaaaga 5041 ccgaggtgca gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgaca 5101 agctgatcgc cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca 5161 ccgtggccta ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga 5221 agagtgtgaa agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc 5281 ccatcgactt tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc 5341 tgcctaagta ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg 5401 gcgaactgca gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc 5461 tggccagcca ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt 5521 ttgtggaaca gcacaaacac tacctggacg agatcatcga gcagatcagc gagttctcca 5581 agagagtgat cctggccgac gctaatctgg acaaggtgct gagcgcctac aacaagcaca 5641 gagacaagcc tatcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc 5701 tgggagcccc tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca 5761 gcaccaaaga ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga 5821 cacggatcga cctgtctcag ctgggaggcg acgcctatcc ctatgacgtg cccgattatg 5881 ccagcctggg cagcggctcc cccaagaaaa aacgcaaggt ggaagatcct aagaaaaagc 5941 ggaaagtgga cggcattggt agtgggagca acggcagcag cggatccaac ggtccgactg 6001 acgccgcgga agaggagctt ctgagcaaaa actatcacct cgaaaacgag gttgcgcgac 6061 tgaagaaagg aagcgggtcc ggtggaagtg gctccggatc tggaggttct ggcagcggag 6121 gtagcggcag tggcgaagag ctccttagta agaactatca tctggaaaat gaggtagcgc 6181 gcttaaagaa agggtcggga agtggcggca gcggaagtgg gagtggaggg agcggttctg 6241 gcggttccgg cagtggagag gagttgctgt ctaagaacta ccacttagaa aacgaagtcg 6301 cacggctaaa aaaaggttcc ggctccggcg gctccggttc tggaagcggg ggctcgggat 6361 caggtggatc tggatcagga gaggaattgc tttccaaaaa ctaccacctt gagaatgagg 6421 tggccaggtt aaagaagggg agcggctcgg ggggtagtgg atcggggtcg ggcgggtcag 6481 gaagcggtgg tagcggatct ggggaggagc tgctctcgaa gaattaccat ttggagaacg 6541 aagtggcgag actaaagaag ggaagcggta gtggtggttc agggtctggt tcaggtggca 6601 gtgggtctgg gggctcaggg tccgggacgg ccggcctcgg aagcggagct actaacttca 6661 gcctgctgaa gcaggctgga gacgtggagg agaaccctgg acctagtacc atgggccccg 6721 acatcgtgat gacccagagc cccagcagcc tgagcgccag cgtgggcgac cgcgtgacca 6781 tcacctgccg cagcagcacc ggcgccgtga ccaccagcaa ctacgccagc tgggtgcagg 6841 agaagcccgg caagctgttc aagggcctga tcggcggcac caacaaccgc gcccccggcg 6901 tgcccagccg cttcagcggc agcctgatcg gcgacaaggc caccctgacc atcagcagcc 6961 tgcagcccga ggacttcgcc acctacttct gcgccctgtg gtacagcaac cactgggtgt 7021 tcggccaggg caccaaggtg gagctgaagc gcggcggcgg cggcagcggc ggcggcggca 7081 gcggcggcgg cggcagcagc ggcggcggca gcgaggtgaa gctgctggag agcggcggcg 7141 gcctggtgca gcccggcggc agcctgaagc tgagctgcgc cgtgagcggc ttcagcctga 7201 ccgactacgg cgtgaactgg gtgcgccagg cccccggccg cggcctggag tggatcggcg 7261 tgatctgggg cgacggcatc accgactaca acagcgccct gaaggaccgc ttcatcatca 7321 gcaaggacaa cggcaagaac accgtgtacc tgcagatgag caaggtgcgc agcgacgaca 7381 ccgccctgta ctactgcgtg accggcctgt tcgactactg gggccagggc accctggtga 7441 ccgtgagcag ctacccatac gatgttccag attacgctgg tggaggcgga ggttctgggg 7501 gaggaggtag tggcggtggt ggttcaggag gcggcggaag cttggatcca ggtggaggtg 7561 gaagcggtag caaaggagaa gaacttttca ctggagttgt cccaattctt gttgaattag 7621 atggtgatgt taatgggcac aaattttctg tccgtggaga gggtgaaggt gatgctacaa 7681 acggaaaact cacccttaaa tttatttgca ctactggaaa actacctgtt ccgtggccaa 7741 cacttgtcac tactctgacc tatggtgttc aatgcttttc ccgttatccg gatcacatga 7801 aacggcatga ctttttcaag agtgccatgc ccgaaggtta tgtacaggaa cgcactatat 7861 ctttcaaaga tgacgggacc tacaagacgc gtgctgaagt caagtttgaa ggtgataccc 7921 ttgttaatcg tatcgagtta aagggtattg attttaaaga agatggaaac attcttggac 7981 acaaactcga gtacaacttt aactcacaca atgtatacat cacggcagac aaacaaaaga 8041 atggaatcaa agctaacttc aaaattcgcc acaacgttga agatggttcc gttcaactag 8101 cagaccatta tcaacaaaat actccaattg gcgatggccc tgtcctttta ccagacaacc 8161 attacctgtc gacacaatct gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg 8221 tccttcttga gtttgtaact gctgctggga ttacacatgg catggatgag ctctacaaag 8281 gtggaggtcg gaccggtggc ggtggcggag gggctagcag atccgaactg cccacctgca 8341 gctgtcttga tcgagttata caaaaagaca aaggcccata ttatacacac cttggggcag 8401 gaccaagtgt tgctgctgtc agggaaatca tggagaatag gtatggtcaa aaaggaaacg 8461 caataaggat agaaatagta gtgtacaccg gtaaagaagg gaaaagctct catgggtgtc 8521 caattgctaa gtgggtttta agaagaagca gtgatgaaga aaaagttctt tgtttggtcc 8581 ggcagcgtac aggccaccac tgtccaactg ctgtgatggt ggtgctcatc atggtgtggg 8641 atggcatccc tcttccaatg gccgaccggc tatacacaga gctcacagag aatctaaagt 8701 catacaatgg gcaccctacc gacagaagat gcaccctcaa tgaaaatcgt acctgtacat 8761 gtcaaggaat tgatccagag acttgtggag cttcattctc ttttggctgt tcatggagta 8821 tgtactttaa tggctgtaag tttggtagaa gcccaagccc cagaagattt agaattgatc 8881 caagctctcc cttacatgaa aaaaaccttg aagataactt acagagtttg gctacacgat 8941 tagctccaat ttataagcag tatgctccag tagcttacca aaatcaggtg gaatatgaaa 9001 atgttgcccg agaatgtcgg cttggcagca aggaaggtcg tcccttctct ggggtcactg 9061 cttgcctgga cttctgtgct catccccaca gggacattca caacatgaat aatggaagca 9121 ctgtggtttg taccttaact cgagaagata accgctcttt gggtgttatt cctcaagatg 9181 agcagctcca tgtgctacct ctttataagc tttcagacac agatgagttt ggctccaagg 9241 aaggaatgga agccaagatc aaatctgggg ccatcgaggt cctggcaccc cgccgcaaaa 9301 aaagaacgtg tttcactcag cctgttcccc gttctggaaa gaagagggct gcgatgatga 9361 cagaggttct tgcacataag ataagggcag tggaaaagaa acctattccc cgaatcaagc 9421 ggaagaataa ctcaacaaca acaaacaaca gtaagccttc gtcactgcca accttaggga 9481 gtaacactga gaccgtgcaa cctgaagtaa aaagtgaaac cgaaccccat tttatcttaa 9541 aaagttcaga caacactaaa acttattcgc tgatgccatc cgctcctcac ccagtgaaag 9601 aggcatctcc aggcttctcc tggtccccga agactgcttc agccacacca gctccactga 9661 agaatgacgc aacagcctca tgcgggtttt cagaaagaag cagcactccc cactgtacga 9721 tgccttcggg aagactcagt ggtgccaatg cagctgctgc tgatggccct ggcatttcac 9781 agcttggcga agtggctcct ctccccaccc tgtctgctcc tgtgatggag cccctcatta 9841 attctgagcc ttccactggt gtgactgagc cgctaacgcc tcatcagcca aaccaccagc 9901 cctccttcct cacctctcct caagaccttg cctcttctcc aatggaagaa gatgagcagc 9961 attctgaagc agatgagcct ccatcagacg aacccctatc tgatgacccc ctgtcacctg 10021 ctgaggagaa attgccccac attgatgagt attggtcaga cagtgagcac atctttttgg 10081 atgcaaatat tggtggggtg gccatcgcac ctgctcacgg ctcggttttg attgagtgtg 10141 cccggcgaga gctgcacgct accactcctg ttgagcaccc caaccgtaat catccaaccc 10201 gcctctccct tgtcttttac cagcacaaaa acctaaataa gccccaacat ggttttgaac 10261 taaacaagat taagtttgag gctaaagaag ctaagaataa gaaaatgaag gcctcagagc 10321 aaaaagacca ggcagctaat gaaggtccag aacagtcctc tgaagtaaat gaattgaacc 10381 aaattccttc tcataaagca ttaacattaa cccatgacaa tgttgtcacc gtgtcccctt 10441 atgctctcac acacgttgcg gggccctata accattgggt ctgagcggcc gcgactctag 10501 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac 10561 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca 10621 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt 10681 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttggcg cgcctgtaca 10741 aaaaagcagg ctttaaagga accaattcag tcgactggat ccggtaccaa ggtcgggcag 10801 gaagagggcc tatttcccat gattccttca tatttgcata tacgatacaa ggctgttaga 10861 gagataatta gaattaattt gactgtaaac acaaagatat tagtacaaaa tacgtgacgt 10921 agaaagtaat aatttcttgg gtagtttgca gttttaaaat tatgttttaa aatggactat 10981 catatgctta ccgtaacttg aaagtatttc gatttcttgg ctttatatat cttaagttaa 11041 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct ttttttctag 11101 acccagcttt cttgtacaaa gttggcatta ggcgcgccaa ggcgtaaatt gtaagcgtta 11161 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg 11221 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg 11281 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa 11341 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg 11401 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt 11461 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg 11521 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta 11581 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta 11641 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat 11701 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa 11761 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag 11821 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag 11881 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc 11941 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt 12001 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg 12061 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt 12121 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc 12181 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc 12241 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg 12301 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag 12361 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg 12421 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg 12481 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac 12541 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg 12601 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc 12661 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg 12721 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc 12781 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc 12841 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc 12901 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc 12961 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg 13021 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt 13081 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac 13141 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca 13201 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg 13261 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg 13321 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata 13381 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct 13441 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga 13501 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg 13561 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc 13621 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct 13681 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc 13741 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt 13801 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg 13861 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct 13921 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag 13981 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag 14041 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg 14101 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg 14161 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac 14221 cgccatgcat tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata 14281 tggagttccg //
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pPlatTETgRNA2载体质粒应用举例
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