质粒名称: | pCS6-StaPLd-Casp9 |
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出品公司: | Addgene |
目录编号: | 111511 |
存储实验室: | Michael Lin |
载体骨架基本信息 |
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载体名称: | pCMV-SPORT6 |
载体出品公司: | Invitrogen |
空载体大小: | -- |
完整质粒大小: | 6801 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: | DH5a |
拷贝数: | -- |
插入基因信息 |
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插入基因名称: | StaPLd-Casp9 |
物种来源: | H. sapiens (human); Hepatitis C Virus (HCV) genotype 1a |
插入大小: | 2457 bp |
基因突变: | HCV NS3蛋白酶携带V36M、T54A和S122G突变。两个胱天蛋白酶-9拷贝的N末端都被截短了。 |
GenBank编号 | NC_004102 |
启动子: | CMV |
标签/融合蛋白: | -- |
克隆方法: | Gibson Cloning |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | ACAGCTATGACCATTAGGCCT |
3’测序引物 | GTTGTAAAACGACGGCCAGT |
文献引用方法 |
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材料和方法部分: | pCS6-StaPLd-Casp9 was a gift from Michael Lin (Addgene plasmid # 111511 ; http://n2t.net/addgene:111511 ; RRID:Addgene_111511) |
参考文献部分: | StaPLs: versatile genetically encoded modules for engineering drug-inducible proteins. Jacobs CL, Badiee RK, Lin MZ. Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z. Epub 2018 Jul 2. 10.1038/s41592-018-0041-z PubMed 29967496 |
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LOCUS pCS6-StaPLd-Casp9 6798 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses a tandem caspase-9 dimer, governed by a StaPL(AI) module, in mammalian cells. [AI = asunaprevir inhibited].. ACCESSION . VERSION . KEYWORDS pCS6-StaPLd-Casp9 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6798) AUTHORS Jacobs CL, Badiee RK, Lin MZ TITLE StaPLs: versatile genetically encoded modules for engineering drug-inducible proteins. JOURNAL Nat Methods. 2018 Jul;15(7):523-526. doi: 10.1038/s41592-018-0041-z. Epub 2018 Jul 2. PUBMED 29967496 REFERENCE 2 (bases 1 to 6798) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6798 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 157..1017 /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 complement(375..394) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(1119..1152) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." rep_origin 1230..1818 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 1719..1738 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 1972..1989 /label=L4440 /note="L4440 vector, forward primer" enhancer 2277..2580 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2581..2784 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2735..2755 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2781..2805 /label=LNCX /note="Human CMV promoter, forward primer" primer_bind 2910..2926 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 2910..2926 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 2937..2955 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2937..2954 /label=SP6 /note="SP6 promoter, forward primer" protein_bind 2956..2980 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" regulatory 2999..3008 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS complement(3326..3337) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" CDS 3872..3898 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" CDS complement(4943..4954) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" protein_bind complement(5478..5502) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" primer_bind complement(5503..5522) /label=T7 /note="T7 promoter, forward primer" promoter complement(5504..5522) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5533..5550) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(5533..5549) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5542..5564) /label=M13/pUC Forward /note="In lacZ gene" polyA_signal 5954..6088 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5991..6010) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6045..6064 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" rep_origin 6214..6668 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6301..6320) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 6510..6531 /label=F1ori-F /note="F1 origin, forward primer" ORIGIN 1 gctggaggga gaataaaggt ctaagatgtg cgatagaggg aagtcgcatt gaattatgtg 61 ctgtgtaggg atcgctggta tcaaatatgt gtgcccaccc ctggcatgag acaataaccc 121 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc 181 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg 241 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat 301 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc 361 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa 421 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa 481 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt 541 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct 601 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat 661 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg 721 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg 781 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt 841 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg 901 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg 961 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg 1021 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 1081 aggatctagg tgaagatcct ttttgataat ctcatgccat aacttcgtat aatgtatgct 1141 atacgaagtt atggcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca 1201 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc 1261 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta 1321 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt 1381 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc 1441 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg 1501 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg 1561 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag 1621 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc 1681 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat 1741 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg 1801 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc 1861 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt 1921 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca 1981 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg 2041 attcattaat gcagagcttg caattcgcgc tttttcaata ttattgaagc atttatcagg 2101 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg 2161 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga 2221 cattaaccta taaaaatagg cgtattacga ggccctttca cgcattagat gcatgtcgtt 2281 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg 2341 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg 2401 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt 2461 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg 2521 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg 2581 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt 2641 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac 2701 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg 2761 tgggaggtct atataagcag agctcgttta gtgaaccgtc agatcgcctg gagacgccat 2821 ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg gactctagcc 2881 taggccgcgg gacggataac aatttcacac aggaaacagc tatgaccatt aggcctattt 2941 aggtgacact atagaacaag tttgtacaaa aaagcaggct ggtaccggtc cggaattcgc 3001 caccatggat gtcggtgctc ttgagagttt gaggggaaat gcagatttgg cttacatcct 3061 gagcatggag ccctgtggcc actgcctcat tatcaacaat gtgaacttct gccgtgagtc 3121 cgggctccgc acccgcactg gctccaacat cgactgtgag aagttgcggc gtcgcttctc 3181 ctcgctgcat ttcatggtgg aggtgaaggg cgacctgact gccaagaaaa tggtgctggc 3241 tttgctggag ctggcgcggc aggaccacgg tgctctggac tgctgcgtgg tggtcattct 3301 ctctcacggc tgtcaggcca gccacctgca gttcccaggg gctgtctacg gcacagatgg 3361 atgccctgtg tcggtcgaga agattgtgaa catcttcaat gggaccagct gccccagcct 3421 gggagggaag cccaagctct ttttcatcca ggcctgtggt ggggagcaga aagaccatgg 3481 gtttgaggtg gcctccactt cccctgaaga cgagtcccct ggcagtaacc ccgagccaga 3541 tgccaccccg ttccaggaag gtttgaggac cttcgaccag ctggacgcca tatctagttt 3601 gcccacaccc agtgacatct ttgtgtccta ctctactttc ccaggttttg tttcctggag 3661 ggaccccaag agtggctcct ggtacgttga gaccctggac gacatctttg agcagtgggc 3721 tcactctgaa gacctgcagt ccctcctgct tagggtcgct aatgctgttt cggtgaaagg 3781 gatttataaa cagatgcctg gttgctttaa tttcctccgg aaaaaacttt tctttaaaac 3841 atcagggggc agtggcggga gcgggcgcgc ctatccctac gatgtgcccg attacgctgg 3901 tggatccggc gggtcagatg agatggaaga gtgctctcag cacggcggat ctggaggttc 3961 tacaggctgc gtggtcatag tgggcaggat cgtcttgtcc ggatccggca ctagtgcgcc 4021 catcacggcg tacgcccagc agacgagagg cctcctaggg tgtataatca ccagcctgac 4081 tggccgggac aaaaaccaag tggagggtga ggtccagatc atgtcaactg ctacccaaac 4141 cttcctggca acgtgcatca atggggtatg ctgggcagtc taccacgggg ccggaacgag 4201 gaccatcgca tcacccaagg gtcctgtcat ccagatgtat accaatgtgg accaagacct 4261 tgtgggctgg cccgctcctc aaggttcccg ctcattgaca ccctgtacct gcggctcctc 4321 ggacctttac ctggtcacga ggcacgccga tgtcattccc gtgcgccggc gaggtgatgg 4381 caggggtagc ctgctttcgc cccggcccat ttcctacttg aaaggctcct cggggggtcc 4441 gctgttgtgc cccgcgggac acgccgtggg cctattcagg gccgcggtgt gcacccgtgg 4501 agtggctaaa gcggtggact ttatccctgt ggagaaccta gagacaacca tgagatcccc 4561 ggtgttcacg gacaactcct ctccaccagc agtcaccctg acgcacgggg gttcaggcgg 4621 tagtgatgtc ggtgctcttg agagtttgag gggaaatgca gatttggctt acatcctgag 4681 catggagccc tgtggccact gcctcattat caacaatgtg aacttctgcc gtgagtccgg 4741 gctccgcacc cgcactggct ccaacatcga ctgtgagaag ttgcggcgtc gcttctcctc 4801 gctgcatttc atggtggagg tgaagggcga cctgactgcc aagaaaatgg tgctggcttt 4861 gctggagctg gcgcggcagg accacggtgc tctggactgc tgcgtggtgg tcattctctc 4921 tcacggctgt caggccagcc acctgcagtt cccaggggct gtctacggca cagatggatg 4981 ccctgtgtcg gtcgagaaga ttgtgaacat cttcaatggg accagctgcc ccagcctggg 5041 agggaagccc aagctctttt tcatccaggc ctgtggtggg gagcagaaag accatgggtt 5101 tgaggtggcc tccacttccc ctgaagacga gtcccctggc agtaaccccg agccagatgc 5161 caccccgttc caggaaggtt tgaggacctt cgaccagctg gacgccatat ctagtttgcc 5221 cacacccagt gacatctttg tgtcctactc tactttccca ggttttgttt cctggaggga 5281 ccccaagagt ggctcctggt acgttgagac cctggacgac atctttgagc agtgggctca 5341 ctctgaagac ctgcagtccc tcctgcttag ggtcgctaat gctgtttcgg tgaaagggat 5401 ttataaacag atgcctggtt gctttaattt cctccggaaa aaacttttct ttaaaacatc 5461 atagaagctt tacgcgtacc cagctttctt gtacaaagtg gtccctatag tgagtcgtat 5521 tataagctag gcactggccg tcgttttaca acgtcgtgac tgggaaaact gctagcttgg 5581 gatctttgtg aaggaacctt acttctgtgg tgtgacataa ttggacaaac tacctacaga 5641 gatttaaagc tctaaggtaa atataaaatt tttaagtgta taatgtgtta aactagctgc 5701 atatgcttgc tgcttgagag ttttgcttac tgagtatgat ttatgaaaat attatacaca 5761 ggagctagtg attctaattg tttgtgtatt ttagattcac agtcccaagg ctcatttcag 5821 gcccctcagt cctcacagtc tgttcatgat cataatcagc cataccacat ttgtagaggt 5881 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc 5941 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat 6001 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact 6061 catcaatgta tcttatcatg tctggatcga tcctgcatta atgaatcggc caacgcgcgg 6121 ggagaggcgg tttgcgtatt ggctggcgta atagcgaaga ggcccgcacc gatcgccctt 6181 cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg 6241 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg 6301 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc 6361 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa 6421 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc 6481 ctttgacgtt ggagtccacg ttcttaatag tggactcttg ttccaaactg gaacaacact 6541 caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg 6601 gttaaaaaat gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct 6661 tacaatttag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc 6721 taaatacatt caaatatgta tccgctcatg ccaggtcttg gactggtgag aacggcttgc 6781 tcggcagctt cgatgtgt //
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