质粒名称: | pX603-AAV-CMV::NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA |
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出品公司: | 爱迪基因 |
目录编号: | 61594 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | pAAV |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳动物表达,AAV,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: | hSaCas9 |
物种来源: | -- |
插入大小: | 3345 bp |
基因突变: | D10A, N580A |
GenBank编号 | CCK74173.1 |
启动子 | -- |
标签/融合蛋白: |
NLS(插入件上的N端子)
NLS(插接件上的C端子)
3xHA(插入件上的C端子)
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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: | pX603-AAV-CMV::NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA was a gift from Feng Zhang (Addgene plasmid # 61594 ; http://n2t.net/addgene:61594 ; RRID:Addgene_61594) |
参考文献部分: | In vivo genome editing using Staphylococcus aureus Cas9. Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F. Nature. 2015 Apr 1. doi: 10.1038/nature14299. 10.1038/nature14299 PubMed 25830891 |
pX603-AAV-CMV::NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA
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 pX603-AAV-CMV::NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA 7090 bp ds-DNA circular SYN 23-AUG-2018 DEFINITION A catalytically inactive SaCas9 (dSaCas9) with the D10A and N580A mutations. . ACCESSION . VERSION . KEYWORDS pX603-AAV-CMV::NLS-dSaCas9(D10A,N580A)-NLS-3xHA-bGHpA SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7090) AUTHORS Ran FA, Cong L, Yan WX, Scott DA, Gootenberg JS, Kriz AJ, Zetsche B, Shalem O, Wu X, Makarova KS, Koonin EV, Sharp PA, Zhang F TITLE In vivo genome editing using Staphylococcus aureus Cas9. JOURNAL Nature. 2015 Apr 1. doi: 10.1038/nature14299. PUBMED 25830891 REFERENCE 2 (bases 1 to 7090) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 23, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7090 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 42..182 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 53..182 /label=AAV2 ITR enhancer 202..581 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 582..785 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 736..756 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" regulatory 804..813 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 816..836 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4017..4064 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" CDS 4071..4151 /codon_start=1 /product="three tandem HA epitope tags" /label=3xHA /translation="YPYDVPDYAYPYDVPDYAYPYDVPDYA" primer_bind complement(4179..4196) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4185..4392 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 4405..4534 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" rep_origin complement(4596..5184) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4676..4695) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5355..6215) /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 5978..5997 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6216..6320) /gene="bla" /label=AmpR promoter primer_bind 6388..6406 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6444..6466) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 6566..6585 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(6602..7057) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6739..6760) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 6951..6970 /label=F1ori-R /note="F1 origin, reverse primer" ORIGIN 1 accgcacaga tgcgtaagga gaaaataccg catcaggcgc ccctgcaggc agctgcgcgc 61 tcgctcgctc actgaggccg cccgggcaaa gcccgggcgt cgggcgacct ttggtcgccc 121 ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc aactccatca ctaggggttc 181 ctgcggccgc ctcgaggcgt tgacattgat tattgactag ttattaatag taatcaatta 241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg 301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc 361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa 421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca 481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta 541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt 601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg 661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca 721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca 781 gagctctctg gctaactacc ggtgccacca tggccccaaa gaagaagcgg aaggtcggta 841 tccacggagt cccagcagcc aagcggaact acatcctggg cctggccatc ggcatcacca 901 gcgtgggcta cggcatcatc gactacgaga cacgggacgt gatcgatgcc ggcgtgcggc 961 tgttcaaaga ggccaacgtg gaaaacaacg agggcaggcg gagcaagaga ggcgccagaa 1021 ggctgaagcg gcggaggcgg catagaatcc agagagtgaa gaagctgctg ttcgactaca 1081 acctgctgac cgaccacagc gagctgagcg gcatcaaccc ctacgaggcc agagtgaagg 1141 gcctgagcca gaagctgagc gaggaagagt tctctgccgc cctgctgcac ctggccaaga 1201 gaagaggcgt gcacaacgtg aacgaggtgg aagaggacac cggcaacgag ctgtccacca 1261 aagagcagat cagccggaac agcaaggccc tggaagagaa atacgtggcc gaactgcagc 1321 tggaacggct gaagaaagac ggcgaagtgc ggggcagcat caacagattc aagaccagcg 1381 actacgtgaa agaagccaaa cagctgctga aggtgcagaa ggcctaccac cagctggacc 1441 agagcttcat cgacacctac atcgacctgc tggaaacccg gcggacctac tatgagggac 1501 ctggcgaggg cagccccttc ggctggaagg acatcaaaga atggtacgag atgctgatgg 1561 gccactgcac ctacttcccc gaggaactgc ggagcgtgaa gtacgcctac aacgccgacc 1621 tgtacaacgc cctgaacgac ctgaacaatc tcgtgatcac cagggacgag aacgagaagc 1681 tggaatatta cgagaagttc cagatcatcg agaacgtgtt caagcagaag aagaagccca 1741 ccctgaagca gatcgccaaa gaaatcctcg tgaacgaaga ggatattaag ggctacagag 1801 tgaccagcac cggcaagccc gagttcacca acctgaaggt gtaccacgac atcaaggaca 1861 ttaccgcccg gaaagagatt attgagaacg ccgagctgct ggatcagatt gccaagatcc 1921 tgaccatcta ccagagcagc gaggacatcc aggaagaact gaccaatctg aactccgagc 1981 tgacccagga agagatcgag cagatctcta atctgaaggg ctataccggc acccacaacc 2041 tgagcctgaa ggccatcaac ctgatcctgg acgagctgtg gcacaccaac gacaaccaga 2101 tcgctatctt caaccggctg aagctggtgc ccaagaaggt ggacctgtcc cagcagaaag 2161 agatccccac caccctggtg gacgacttca tcctgagccc cgtcgtgaag agaagcttca 2221 tccagagcat caaagtgatc aacgccatca tcaagaagta cggcctgccc aacgacatca 2281 ttatcgagct ggcccgcgag aagaactcca aggacgccca gaaaatgatc aacgagatgc 2341 agaagcggaa ccggcagacc aacgagcgga tcgaggaaat catccggacc accggcaaag 2401 agaacgccaa gtacctgatc gagaagatca agctgcacga catgcaggaa ggcaagtgcc 2461 tgtacagcct ggaagccatc cctctggaag atctgctgaa caaccccttc aactatgagg 2521 tggaccacat catccccaga agcgtgtcct tcgacaacag cttcaacaac aaggtgctcg 2581 tgaagcagga agaagccagc aagaagggca accggacccc attccagtac ctgagcagca 2641 gcgacagcaa gatcagctac gaaaccttca agaagcacat cctgaatctg gccaagggca 2701 agggcagaat cagcaagacc aagaaagagt atctgctgga agaacgggac atcaacaggt 2761 tctccgtgca gaaagacttc atcaaccgga acctggtgga taccagatac gccaccagag 2821 gcctgatgaa cctgctgcgg agctacttca gagtgaacaa cctggacgtg aaagtgaagt 2881 ccatcaatgg cggcttcacc agctttctgc ggcggaagtg gaagtttaag aaagagcgga 2941 acaaggggta caagcaccac gccgaggacg ccctgatcat tgccaacgcc gatttcatct 3001 tcaaagagtg gaagaaactg gacaaggcca aaaaagtgat ggaaaaccag atgttcgagg 3061 aaaagcaggc cgagagcatg cccgagatcg aaaccgagca ggagtacaaa gagatcttca 3121 tcacccccca ccagatcaag cacattaagg acttcaagga ctacaagtac agccaccggg 3181 tggacaagaa gcctaataga gagctgatta acgacaccct gtactccacc cggaaggacg 3241 acaagggcaa caccctgatc gtgaacaatc tgaacggcct gtacgacaag gacaatgaca 3301 agctgaaaaa gctgatcaac aagagccccg aaaagctgct gatgtaccac cacgaccccc 3361 agacctacca gaaactgaag ctgattatgg aacagtacgg cgacgagaag aatcccctgt 3421 acaagtacta cgaggaaacc gggaactacc tgaccaagta ctccaaaaag gacaacggcc 3481 ccgtgatcaa gaagattaag tattacggca acaaactgaa cgcccatctg gacatcaccg 3541 acgactaccc caacagcaga aacaaggtcg tgaagctgtc cctgaagccc tacagattcg 3601 acgtgtacct ggacaatggc gtgtacaagt tcgtgaccgt gaagaatctg gatgtgatca 3661 aaaaagaaaa ctactacgaa gtgaatagca agtgctatga ggaagctaag aagctgaaga 3721 agatcagcaa ccaggccgag tttatcgcct ccttctacaa caacgatctg atcaagatca 3781 acggcgagct gtatagagtg atcggcgtga acaacgacct gctgaaccgg atcgaagtga 3841 acatgatcga catcacctac cgcgagtacc tggaaaacat gaacgacaag aggcccccca 3901 ggatcattaa gacaatcgcc tccaagaccc agagcattaa gaagtacagc acagacattc 3961 tgggcaacct gtatgaagtg aaatctaaga agcaccctca gatcatcaaa aagggcaaaa 4021 ggccggcggc cacgaaaaag gccggccagg caaaaaagaa aaagggatcc tacccatacg 4081 atgttccaga ttacgcttat ccctacgacg tgcctgatta tgcataccca tatgatgtcc 4141 ccgactatgc ctaagaattc ctagagctcg ctgatcagcc tcgactgtgc cttctagttg 4201 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 4261 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 4321 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag agaatagcag 4381 gcatgctggg gagctagagg ccgcaggaac ccctagtgat ggagttggcc actccctctc 4441 tgcgcgctcg ctcgctcact gaggccgggc gaccaaaggt cgcccgacgc ccgggcggcc 4501 tcagtgagcg agcgagcgcg cagctgcctg caggacatgt gagcaaaagg ccagcaaaag 4561 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4621 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4681 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4741 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4801 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4861 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 4921 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 4981 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5041 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5101 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5161 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5221 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5281 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5341 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5401 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5461 ttaccatctg gccccagtgc tgcaatgata ccgcggcttc cacgctcacc ggctccagat 5521 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5581 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5641 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5701 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5761 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5821 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 5881 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 5941 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6001 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6061 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6121 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6181 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6241 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6301 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc 6361 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg 6421 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct 6481 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc 6541 gggtgtcggg gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat 6601 aaaattgtaa acgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca 6661 ttttttaacc aatagaccga aatcggcaaa atcccttata aatcaaaaga atagcccgag 6721 atagagttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc 6781 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc 6841 aaatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc 6901 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa 6961 gcgaaaggag cgggcgctaa ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc 7021 acacccgccg cgcttaatgc gccgctacag ggcgcgtact atggttgctt tgacgtatgc 7081 ggtgtgaaat //