质粒名称: | pDF0159 pCMV - huDisCas7-11 mammalian expression |
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出品公司: | Addgene |
目录编号: | 172507 |
存储实验室: | Omar Abudayyeh, Jonathan Gootenberg |
载体骨架基本信息 |
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载体名称: | Unspecified |
载体出品公司: | -- |
完整质粒大小: | 8205 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达 |
筛选标记: | -- |
载体抗性: | 氨苄青霉素,100μg/mL |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | -- |
插入基因信息 |
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插入基因名称: | pCMV-huDisCas7-11哺乳动物表达 |
物种来源: | 石本脱硫虫 |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子: | -- |
标签/融合蛋白: | -- |
克隆方法: | Gibson Cloning |
文献引用方法 |
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材料和方法部分: | pDF0159 pCMV - huDisCas7-11 mammalian expression was a gift from Omar Abudayyeh & Jonathan Gootenberg (Addgene plasmid # 172507 ; http://n2t.net/addgene:172507 ; RRID:Addgene_172507) |
参考文献部分: | Programmable RNA targeting with the single-protein CRISPR effector Cas7-11. Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, Koonin EV, Abudayyeh OO, Gootenberg JS. Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 10.1038/s41586-021-03886-5. 10.1038/s41586-021-03886-5 PubMed 34489594 |
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LOCUS pDF0159 pCMV - huDisCas7-11 mammalian expression 8205 bp ds-DNA circular SYN 07-OCT-2021 DEFINITION Encodes human codon optimized DisCas7-11 protein alone for expression in mammalian cells. ACCESSION . VERSION . KEYWORDS pDF0159 pCMV - huDisCas7-11 mammalian expression SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8205) AUTHORS Ozcan A, Krajeski R, Ioannidi E, Lee B, Gardner A, Makarova KS, Koonin EV, Abudayyeh OO, Gootenberg JS TITLE Programmable RNA targeting with the single-protein CRISPR effector Cas7-11. JOURNAL Nature. 2021 Sep 6. pii: 10.1038/s41586-021-03886-5. doi: 10.1038/s41586-021-03886-5. PUBMED 34489594 REFERENCE 2 (bases 1 to 8205) AUTHORS . TITLE Direct Submission JOURNAL Exported Oct 7, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..8205 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(70..89) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 261..640 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 641..844 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 795..815 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 886..905 /label=T7 /note="T7 promoter, forward primer" promoter 886..904 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 5729..5746 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" primer_bind complement(5769..5786) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 5775..5999 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(6070..6086) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6070..6086) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6083..6105) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6094..6110 /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(6118..6148) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6163..6184 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6301..6318) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6472..7060) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6552..6571) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7231..8091) /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 7854..7873 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8092..8196) /gene="bla" /label=AmpR promoter ORIGIN 1 ccgaaaagtg ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc 61 tactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg 121 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt 181 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt 241 acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac 301 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg 361 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 421 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 481 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 541 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 601 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 661 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 721 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa 781 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 841 agctggttta gtgaaccgtc agatccgcta gagatccgcg gccgctaata cgactcacta 901 tagggagagc cgccaccatg acgactacta tgaagatttc aattgaattc ctcgagccgt 961 ttcggatgac caaatggcag gaaagcacta ggcgaaacaa gaacaacaag gaatttgttc 1021 gcggccaagc ttttgcaaga tggcatagaa ataagaagga taacacaaaa gggcggcctt 1081 acataactgg gacattgctg cggtccgcag taattagaag cgcggaaaac ctgctgacat 1141 tgagtgatgg gaaaattagc gaaaagacat gttgtcctgg gaaatttgat acggaagata 1201 aagacaggtt gctgcaattg aggcagcggt ccacccttcg ttggaccgat aagaacccct 1261 gtccagataa cgctgaaact tactgcccgt tttgcgaact tctcgggaga tccggtaacg 1321 atggaaagaa ggctgagaag aaggattggc gctttcgtat tcattttggc aatctttccc 1381 ttccagggaa gcccgatttc gacgggccaa aagctatagg cagccaacgg gtacttaaca 1441 gggtcgattt caaatccggt aaggcccatg acttctttaa ggcttacgaa gtcgaccata 1501 ctaggtttcc ccgcttcgaa ggggagatta ccatagataa taaagtcagc gctgaagcca 1561 ggaaactgct ctgtgattct ctcaagttta cggatcggct gtgtggagct ctgtgcgtaa 1621 tcaggttcga tgaatataca ccagcagccg atagtgggaa acaaaccgaa aatgtccaag 1681 cagaaccgaa tgctaatctc gctgaaaaga ccgcagagca aattattagc attctggacg 1741 ataataagaa aaccgaatat acccgcctgc ttgctgatgc tattcgttca ctccgccgct 1801 ctagtaaact tgtcgctggc ttgccaaagg atcatgacgg gaaggatgac cattacctct 1861 gggatatcgg caagaagaag aaagacgaaa actctgtcac tattaggcaa atccttacga 1921 cctcagcaga caccaaggaa ctcaagaatg ccggaaaatg gagagaattc tgcgagaagc 1981 tgggtgaagc gctgtacctc aagagtaaag acatgagtgg cggcctgaaa attactcgga 2041 gaatactggg cgatgctgaa ttccatggaa agcccgatcg gcttgaaaag agccggtctg 2101 tgtcaatcgg atctgtgttg aaagaaactg tggtatgcgg cgaactggtc gctaaaacgc 2161 ccttcttctt cggagcgata gacgaagatg caaaacaaac cgacctgcaa gtactcctca 2221 ctcccgataa caagtataga ctgccaagaa gcgccgtgcg aggtatactc cgtcgagatc 2281 ttcaaaccta ttttgatagc ccatgcaatg ctgagttggg tggacggcca tgcatgtgta 2341 agacgtgtag gattatgaga gggatcacgg tgatggatgc gcgcagtgag tacaatgccc 2401 cgccagaaat aaggcatcgt acccgcatta atcccttcac aggcacggtt gcggaaggtg 2461 ccctgtttaa tatggaagta gcccccgagg ggattgtctt tccatttcaa ctccggtacc 2521 ggggctctga agatgggctg cccgatgcac tgaaaacggt gttgaaatgg tgggctgagg 2581 ggcaggcatt catgagtggc gctgcctcaa ccgggaaggg ccgattccgg atggaaaatg 2641 ctaaatatga aacgctggat ctgagcgacg agaatcaaag gaatgactat cttaagaatt 2701 ggggatggcg tgacgagaag gggctcgagg aactgaagaa acgactgaac tcaggtctgc 2761 cagagcccgg taattatagg gatccaaaat ggcacgagat taacgtttcc attgagatgg 2821 caagcccttt tattaatggc gacccaatcc gcgcagccgt ggacaaacgt ggtacagatg 2881 tggttacctt cgttaagtat aaagctgaag gggaagaggc gaaacccgta tgtgcataca 2941 aggccgaatc ttttagaggg gtgatcagaa gtgccgtggc acgcattcat atggaagatg 3001 gcgtcccttt gactgagttg actcacagtg actgtgaatg tctcctgtgc caaatctttg 3061 gaagtgagta tgaagccggc aaaataaggt ttgaagatct cgtattcgaa agtgacccgg 3121 aacctgtgac cttcgatcat gtggccatcg atagattcac tggcggtgca gctgataaga 3181 agaaattcga tgattcccct ctgcccggta gccctgcaag accgctcatg ttgaaaggct 3241 ccttctggat ccgcagggac gttctcgagg acgaagagta ctgtaaggca ctcggtaagg 3301 ctcttgcaga tgtgaataat ggcctttatc ccctcggtgg aaagagcgcc atcggctacg 3361 gacaggtcaa gagtctgggt ataaagggag atgataagag gatttctcgc ctcatgaatc 3421 ctgcctttga tgagacagat gtagccgttc cagaaaagcc caaaactgat gccgaggttc 3481 gcatcgaggc agagaaagta tattacccac actatttcgt cgaaccccat aagaaggtgg 3541 aacgcgagga gaaaccctgt ggtcatcaaa agttccacga ggggcgactg acaggtaaaa 3601 ttcggtgtaa gctcattacc aagacacccc tcatcgtccc agatactagt aatgacgatt 3661 tcttcagacc tgcggataaa gaagctcgga aggaaaagga cgaatatcat aaatcatatg 3721 ctttcttcag acttcataaa caaatcatga ttcccgggag cgaattgaga ggaatggtga 3781 gtagtgtcta cgaaactgtg acaaattctt gcttcaggat atttgatgag actaaacggt 3841 tgtcatggcg gatggatgct gatcaccaga atgtgctgca agactttctc ccaggtcgag 3901 tgaccgccga tgggaaacat atacaaaagt tttccgaaac tgcaagggtg cctttctatg 3961 acaaaacgca gaaacatttt gacattcttg atgagcaaga aattgctggt gagaaacctg 4021 ttcggatgtg ggtcaagcgt tttattaaac gactgagcct cgttgatcct gctaaacacc 4081 cccagaagaa acaagacaat aaatggaaaa gacgcaaaga aggcatcgcc acatttatag 4141 agcaaaagaa tggctcttat tattttaacg tggtcaccaa taatggctgc acttctttcc 4201 acttgtggca taaacctgac aattttgacc aggagaaact cgaaggcata cagaatggtg 4261 agaaactgga ttgctgggta agagatagta gataccaaaa ggcctttcaa gagatacccg 4321 agaacgaccc agacggatgg gagtgtaaag agggctacct tcatgtcgtc ggccccagca 4381 aagtagagtt tagtgacaag aaaggggatg tgatcaataa ctttcaagga acactcccat 4441 cagttcccaa cgactggaaa acaattagga cgaacgattt taagaatagg aaaaggaaga 4501 atgaacctgt gttttgttgt gaggatgata agggcaatta ctataccatg gctaaatatt 4561 gcgaaacctt cttcttcgat ctgaaggaga acgaggaata cgagatcccc gagaaagcaa 4621 gaatcaaata caaagaactg ctcagggtct ataacaacaa tcctcaagca gtgccggaga 4681 gcgtatttca gtctagagtt gcccgggaaa acgtggaaaa gctgaagtcc ggagatcttg 4741 tgtatttcaa acataatgaa aagtacgtag aggacatcgt cccagtgcgg atttcccgaa 4801 ctgtagacga taggatgatc ggcaaacgta tgagcgccga tctgcggccg tgccatggag 4861 attgggtgga agatggtgat ctcagtgcct tgaatgcata tcccgagaaa agactcctct 4921 tgcgccaccc caaaggactc tgccctgctt gccggctctt tggaaccgga tcttacaagg 4981 gcagagtcag gtttggattc gcgtcactcg aaaacgatcc ggagtggctg atcccaggca 5041 agaatcccgg cgatccgttt cacggcgggc cggtgatgct ctcattgttg gaacggcctc 5101 gcccgacttg gagtataccg ggatccgaca ataagtttaa agtgcctggc agaaagtttt 5161 acgtccacca ccacgcctgg aaaaccatta aggacgggaa ccatcccaca acaggcaaag 5221 ctattgaaca aagccctaat aaccgcactg tagaagctct cgccggcggg aattccttta 5281 gcttcgaaat tgcctttgag aacctgaaag aatgggagct gggtttgctc atccacagcc 5341 tgcaactcga aaagggtctg gcgcataaac ttggaatggc aaagtctatg ggatttggtt 5401 cagttgaaat tgacgtcgaa tcagtgcgcc tgagaaaaga ttggaagcaa tggcggaatg 5461 gcaattccga aattcccaac tggttgggaa aaggatttgc taaactgaag gaatggttcc 5521 gggacgagct cgattttata gaaaatctta agaaacttct ttggtttcct gagggcgacc 5581 aagcaccccg ggtttgctac cccatgctgc gaaagaagga cgatcctaat gggaatagcg 5641 gttacgaaga actcaaagac ggggaattca agaaagaaga tcggcagaag aagctgacca 5701 cgccgtggac accgtgggca taaccggtca tcatcaccat caccattgag tttaaacccg 5761 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt 5821 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgagaaaat 5881 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag 5941 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc 6001 ttctgaggcg gaaagaacca gctggggctc gataccgtcg acctctagct agagcttggc 6061 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa 6121 catacgagcc ggaagcataa agtgtaaagc ctagggtgcc taatgagtga gctaactcac 6181 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca 6241 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc 6301 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc 6361 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc 6421 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 6481 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 6541 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6601 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6661 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6721 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6781 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6841 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6901 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6961 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 7021 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 7081 tacggggtct gacactcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 7141 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 7201 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 7261 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 7321 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 7381 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 7441 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 7501 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 7561 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7621 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7681 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7741 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7801 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7861 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7921 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7981 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 8041 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 8101 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga 8161 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttcc //
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