质粒名称: | pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) |
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出品公司: | 爱迪基因 |
目录编号: | 42335 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | pUC ori vector |
载体抗性: | 氨苄霉素 |
载体类型: | 哺乳细胞表达 |
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: | humanized S. pyogenes Cas9 (D10A) nickase |
插入大小: | 4272 bp |
基因突变: | D10A nickase-converting mutation |
标签/融合蛋白: | HA(N端) |
文献引用方法 |
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材料和方法部分: | pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) was a gift from Vladislav Verkhusha (Addgene plasmid # 45465) |
参考文献部分: |
Near-infrared fluorescent proteins for multicolor in vivo imaging. Shcherbakova DM, Verkhusha VV. Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521. 10.1038/nmeth.2521 PubMed 23770755 |
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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 pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) 8434 bp DNA SYN DEFINITION pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A) ACCESSION KEYWORDS SOURCE ORGANISM other sequences; artificial sequences; vectors. FEATURES Location/Qualifiers source 1..8434 /organism="pX335-U6-Chimeric_BB-CBh-hSpCas9n(D10A)" /mol_type="other DNA" promoter 10..242 /label="hU6_promoter" misc_feature 172..191 /label="LKO1_5_primer" misc_feature 444..713 /label="CAG_enhancer" CDS complement(1230..3830) /label="ORF frame 3" CDS 1251..5450 /label="ORF frame 3" misc_feature 1254..1280 /label="HA" misc_feature 1284..1304 /label="SV40 NLS" misc_feature 1290..1304 /label="NLS" misc_feature complement(5475..5492) /label="BGH_rev_primer" terminator 5478..5681 /label="bGH_PA_terminator" rep_origin 5929..6235 /label="f1_origin" misc_feature 6503..6525 /label="pGEX_3_primer" promoter 6684..6712 /label="AmpR_promoter" gene 6754..7614 /label="Ampicillin" /gene="Ampicillin" CDS 6754..7614 /label="ORF frame 1" rep_origin 7769..8388 /label="pBR322_origin" ORIGIN 1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag 61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga 121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat 181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt GTGGAAAGGA 241 CGAAACACCg gGTCTTCgaG AAGACctgtt ttagagctaG AAAtagcaag ttaaaataag 301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcTTTTTTg ttttagagct 361 agaaatagca agttaaaata aggctagtcc gtTTTTagcg cgtgcgccaa ttctgcagac 421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc 481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc 541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tGtgcccagt 661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac 781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg 841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga 901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc 961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc 1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg 1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag 1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac 1201 ctggagcacc tgcctgaaat cacttttttt caggttGGac cggtgccacc ATGTACCCAT 1261 ACGATGTTCC AGATTACGCT TCGCCGAAGA AAAAGCGCAA GGTCGAAGCG TCCGACAAGA 1321 AGTACAGCAT CGGCCTGGCC ATCGGCACCA ACTCTGTGGG CTGGGCCGTG ATCACCGACG 1381 AGTACAAGGT GCCCAGCAAG AAATTCAAGG TGCTGGGCAA CACCGACCGG CACAGCATCA 1441 AGAAGAACCT GATCGGAGCC CTGCTGTTCG ACAGCGGCGA AACAGCCGAG GCCACCCGGC 1501 TGAAGAGAAC CGCCAGAAGA AGATACACCA GACGGAAGAA CCGGATCTGC TATCTGCAAG 1561 AGATCTTCAG CAACGAGATG GCCAAGGTGG ACGACAGCTT CTTCCACAGA CTGGAAGAGT 1621 CCTTCCTGGT GGAAGAGGAT AAGAAGCACG AGCGGCACCC CATCTTCGGC AACATCGTGG 1681 ACGAGGTGGC CTACCACGAG AAGTACCCCA CCATCTACCA CCTGAGAAAG AAACTGGTGG 1741 ACAGCACCGA CAAGGCCGAC CTGCGGCTGA TCTATCTGGC CCTGGCCCAC ATGATCAAGT 1801 TCCGGGGCCA CTTCCTGATC GAGGGCGACC TGAACCCCGA CAACAGCGAC GTGGACAAGC 1861 TGTTCATCCA GCTGGTGCAG ACCTACAACC AGCTGTTCGA GGAAAACCCC ATCAACGCCA 1921 GCGGCGTGGA CGCCAAGGCC ATCCTGTCTG CCAGACTGAG CAAGAGCAGA CGGCTGGAAA 1981 ATCTGATCGC CCAGCTGCCC GGCGAGAAGA AGAATGGCCT GTTCGGCAAC CTGATTGCCC 2041 TGAGCCTGGG CCTGACCCCC AACTTCAAGA GCAACTTCGA CCTGGCCGAG GATGCCAAAC 2101 TGCAGCTGAG CAAGGACACC TACGACGACG ACCTGGACAA CCTGCTGGCC CAGATCGGCG 2161 ACCAGTACGC CGACCTGTTT CTGGCCGCCA AGAACCTGTC CGACGCCATC CTGCTGAGCG 2221 ACATCCTGAG AGTGAACACC GAGATCACCA AGGCCCCCCT GAGCGCCTCT ATGATCAAGA 2281 GATACGACGA GCACCACCAG GACCTGACCC TGCTGAAAGC TCTCGTGCGG CAGCAGCTGC 2341 CTGAGAAGTA CAAAGAGATT TTCTTCGACC AGAGCAAGAA CGGCTACGCC GGCTACATTG 2401 ACGGCGGAGC CAGCCAGGAA GAGTTCTACA AGTTCATCAA GCCCATCCTG GAAAAGATGG 2461 ACGGCACCGA GGAACTGCTC GTGAAGCTGA ACAGAGAGGA CCTGCTGCGG AAGCAGCGGA 2521 CCTTCGACAA CGGCAGCATC CCCCACCAGA TCCACCTGGG AGAGCTGCAC GCCATTCTGC 2581 GGCGGCAGGA AGATTTTTAC CCATTCCTGA AGGACAACCG GGAAAAGATC GAGAAGATCC 2641 TGACCTTCCG CATCCCCTAC TACGTGGGCC CTCTGGCCAG GGGAAACAGC AGATTCGCCT 2701 GGATGACCAG AAAGAGCGAG GAAACCATCA CCCCCTGGAA CTTCGAGGAA GTGGTGGACA 2761 AGGGCGCTTC CGCCCAGAGC TTCATCGAGC GGATGACCAA CTTCGATAAG AACCTGCCCA 2821 ACGAGAAGGT GCTGCCCAAG CACAGCCTGC TGTACGAGTA CTTCACCGTG TATAACGAGC 2881 TGACCAAAGT GAAATACGTG ACCGAGGGAA TGAGAAAGCC CGCCTTCCTG AGCGGCGAGC 2941 AGAAAAAGGC CATCGTGGAC CTGCTGTTCA AGACCAACCG GAAAGTGACC GTGAAGCAGC 3001 TGAAAGAGGA CTACTTCAAG AAAATCGAGT GCTTCGACTC CGTGGAAATC TCCGGCGTGG 3061 AAGATCGGTT CAACGCCTCC CTGGGCACAT ACCACGATCT GCTGAAAATT ATCAAGGACA 3121 AGGACTTCCT GGACAATGAG GAAAACGAGG ACATTCTGGA AGATATCGTG CTGACCCTGA 3181 CACTGTTTGA GGACAGAGAG ATGATCGAGG AACGGCTGAA AACCTATGCC CACCTGTTCG 3241 ACGACAAAGT GATGAAGCAG CTGAAGCGGC GGAGATACAC CGGCTGGGGC AGGCTGAGCC 3301 GGAAGCTGAT CAACGGCATC CGGGACAAGC AGTCCGGCAA GACAATCCTG GATTTCCTGA 3361 AGTCCGACGG CTTCGCCAAC AGAAACTTCA TGCAGCTGAT CCACGACGAC AGCCTGACCT 3421 TTAAAGAGGA CATCCAGAAA GCCCAGGTGT CCGGCCAGGG CGATAGCCTG CACGAGCACA 3481 TTGCCAATCT GGCCGGCAGC CCCGCCATTA AGAAGGGCAT CCTGCAGACA GTGAAGGTGG 3541 TGGACGAGCT CGTGAAAGTG ATGGGCCGGC ACAAGCCCGA GAACATCGTG ATCGAAATGG 3601 CCAGAGAGAA CCAGACCACC CAGAAGGGAC AGAAGAACAG CCGCGAGAGA ATGAAGCGGA 3661 TCGAAGAGGG CATCAAAGAG CTGGGCAGCC AGATCCTGAA AGAACACCCC GTGGAAAACA 3721 CCCAGCTGCA GAACGAGAAG CTGTACCTGT ACTACCTGCA GAATGGGCGG GATATGTACG 3781 TGGACCAGGA ACTGGACATC AACCGGCTGT CCGACTACGA TGTGGACCAT ATCGTGCCTC 3841 AGAGCTTTCT GAAGGACGAC TCCATCGACA ACAAGGTGCT GACCAGAAGC GACAAGAACC 3901 GGGGCAAGAG CGACAACGTG CCCTCCGAAG AGGTCGTGAA GAAGATGAAG AACTACTGGC 3961 GGCAGCTGCT GAACGCCAAG CTGATTACCC AGAGAAAGTT CGACAATCTG ACCAAGGCCG 4021 AGAGAGGCGG CCTGAGCGAA CTGGATAAGG CCGGCTTCAT CAAGAGACAG CTGGTGGAAA 4081 CCCGGCAGAT CACAAAGCAC GTGGCACAGA TCCTGGACTC CCGGATGAAC ACTAAGTACG 4141 ACGAGAATGA CAAGCTGATC CGGGAAGTGA AAGTGATCAC CCTGAAGTCC AAGCTGGTGT 4201 CCGATTTCCG GAAGGATTTC CAGTTTTACA AAGTGCGCGA GATCAACAAC TACCACCACG 4261 CCCACGACGC CTACCTGAAC GCCGTCGTGG GAACCGCCCT GATCAAAAAG TACCCTAAGC 4321 TGGAAAGCGA GTTCGTGTAC GGCGACTACA AGGTGTACGA CGTGCGGAAG ATGATCGCCA 4381 AGAGCGAGCA GGAAATCGGC AAGGCTACCG CCAAGTACTT CTTCTACAGC AACATCATGA 4441 ACTTTTTCAA GACCGAGATT ACCCTGGCCA ACGGCGAGAT CCGGAAGCGG CCTCTGATCG 4501 AGACAAACGG CGAAACCGGG GAGATCGTGT GGGATAAGGG CCGGGATTTT GCCACCGTGC 4561 GGAAAGTGCT GAGCATGCCC CAAGTGAATA TCGTGAAAAA GACCGAGGTG CAGACAGGCG 4621 GCTTCAGCAA AGAGTCTATC CTGCCCAAGA GGAACAGCGA TAAGCTGATC GCCAGAAAGA 4681 AGGACTGGGA CCCTAAGAAG TACGGCGGCT TCGACAGCCC CACCGTGGCC TATTCTGTGC 4741 TGGTGGTGGC CAAAGTGGAA AAGGGCAAGT CCAAGAAACT GAAGAGTGTG AAAGAGCTGC 4801 TGGGGATCAC CATCATGGAA AGAAGCAGCT TCGAGAAGAA TCCCATCGAC TTTCTGGAAG 4861 CCAAGGGCTA CAAAGAAGTG AAAAAGGACC TGATCATCAA GCTGCCTAAG TACTCCCTGT 4921 TCGAGCTGGA AAACGGCCGG AAGAGAATGC TGGCCTCTGC CGGCGAACTG CAGAAGGGAA 4981 ACGAACTGGC CCTGCCCTCC AAATATGTGA ACTTCCTGTA CCTGGCCAGC CACTATGAGA 5041 AGCTGAAGGG CTCCCCCGAG GATAATGAGC AGAAACAGCT GTTTGTGGAA CAGCACAAGC 5101 ACTACCTGGA CGAGATCATC GAGCAGATCA GCGAGTTCTC CAAGAGAGTG ATCCTGGCCG 5161 ACGCTAATCT GGACAAAGTG CTGTCCGCCT ACAACAAGCA CCGGGATAAG CCCATCAGAG 5221 AGCAGGCCGA GAATATCATC CACCTGTTTA CCCTGACCAA TCTGGGAGCC CCTGCCGCCT 5281 TCAAGTACTT TGACACCACC ATCGACCGGA AGAGGTACAC CAGCACCAAA GAGGTGCTGG 5341 ACGCCACCCT GATCCACCAG AGCATCACCG GCCTGTACGA GACACGGATC GACCTGTCTC 5401 AGCTGGGAGG CGACAGCCCC AAGAAGAAGA GAAAGGTGGA GGCCAGCtaa gaattcCTAG 5461 AGCTCGCTGA TCAGCCTCGA CTGTGCCTTC TAGTTGCCAG CCATCTGTTG TTTGCCCCTC 5521 CCCCGTGCCT TCCTTGACCC TGGAAGGTGC CACTCCCACT GTCCTTTCCT AATAAAATGA 5581 GGAAATTGCA TCGCATTGTC TGAGTAGGTG TCATTCTATT CTGGGGGGTG GGGTGGGGCA 5641 GGACAGCAAG GGGGAGGATT GGGAAGAgAA TAGCAGGCAT GCTGGGGAgc ggccgcagga 5701 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg 5761 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc 5821 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat 5881 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg 5941 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct 6001 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta 6061 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa 6121 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct 6181 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc 6241 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg 6301 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt 6361 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc 6421 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct 6481 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca 6541 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg 6601 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct 6661 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga 6721 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc 6781 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg 6841 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc 6901 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact 6961 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc 7021 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag 7081 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat 7141 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt 7201 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa 7261 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc 7321 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg 7381 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt 7441 gctgataaat ctggagccgg tgagcgtgga agccgcggta tcattgcagc actggggcca 7501 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat 7561 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca 7621 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 7681 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 7741 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 7801 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 7861 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 7921 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 7981 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 8041 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 8101 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 8161 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 8221 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 8281 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 8341 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 8401 ttcctggcct tttgctggcc ttttgctcac atgt //