质粒名称: | pCMV-Flag-YAP-5SA/S94A<质粒图谱.png/td> |
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出品公司: | Addgene |
目录编号: | 33103 |
存储实验室: | Kunliang Guan |
载体骨架基本信息 |
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载体名称: | pCMV |
载体出品公司: | Clontech |
空载体大小: | 4500 bp |
完整质粒大小: | 6231 bp |
载体类型: | 哺乳细胞表达 |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 未知 |
插入基因信息 |
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插入基因名称: | YAP |
基因突变: | S61A, S94A, S109A, S127A, S164A, S381A |
标签/融合蛋白: | Flag (N terminal on insert) |
文献引用方法 |
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材料和方法部分: | pCMV-Flag-YAP-5SA/S94A was a gift from Kunliang Guan (Addgene plasmid # 33103 ; http://n2t.net/addgene:33103 ; RRID:Addgene_33103) |
参考文献部分: | TEAD mediates YAP-dependent gene induction and growth control. Zhao B, Ye X, Yu J, Li L, Li W, Li S, Yu J, Lin JD, Wang CY, Chinnaiyan AM, Lai ZC, Guan KL. Genes Dev. 2008 Jul 15. 22(14):1962-71. 10.1101/gad.1664408 PubMed 18579750 |
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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 pCMV-Flag-YAP-5SA/S94A 6231 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pCMV-Flag-YAP-5SA/S94A SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6231) AUTHORS Zhao B, Ye X, Yu J, Li L, Li W, Li S, Yu J, Lin JD, Wang CY, Chinnaiyan AM, Lai ZC, Guan KL TITLE TEAD mediates YAP-dependent gene induction and growth control. JOURNAL Genes Dev. 2008 Jul 15. 22(14):1962-71. PUBMED 18579750 REFERENCE 2 (bases 1 to 6231) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..6231 /organism="synthetic DNA construct" /mol_type="other DNA" rep_origin 1..456 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(88..107) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 298..319 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 582..604 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 596..613 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 597..613 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" enhancer 774..1153 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1154..1357 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1308..1328 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" CDS 1391..1414 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" regulatory 1439..1448 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 1448..1471 /codon_start=1 /product="FLAG(R) epitope tag, followed by an enterokinase cleavage site" /label=FLAG /translation="DYKDDDDK" polyA_signal 3028..3650 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3227..3246) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" primer_bind complement(3674..3694) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 3679..4008 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3859..3994 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3921..3940 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind complement(4046..4065) /label=T7 /note="T7 promoter, forward primer" promoter complement(4047..4065) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(4079..4095) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4079..4095) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4092..4114) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4103..4119 /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(4127..4157) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4172..4193 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4310..4327) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4481..5069) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4561..4580) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5240..6100) /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 5863..5882 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6101..6205) /gene="bla" /label=AmpR promoter ORIGIN 1 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg 61 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca 121 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta 181 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc 241 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg 301 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat 361 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta 421 acgcgaattt taacaaaata ttaacgctta caatttccat tcgccattca ggctgcgcaa 481 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg 541 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa 601 aacgacggcc agtgccaagc tgatctatac attgaatcaa tattggcaat tagccatatt 661 agtcattggt tatatagcat aaatcaatat tggctattgg ccattgcata cgttgtatct 721 atatcataat atgtacattt atattggctc atgtccaata tgaccgccat gttgacatcg 781 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat 841 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc 901 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca 961 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta 1021 tcatatgcca agtccgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta 1081 tgcccagtac atgaccttac gggactttcc tacttggcag tacatctacg tattagtcat 1141 cgctattacc atggtgatgc ggttttggca gtacaccaat gggcgtggat agcggtttga 1201 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca 1261 aaatcaacgg gactttccaa aatgtcgtaa taaccccgcc ccgttgacgc aaatgggcgg 1321 taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc gtcagaattg 1381 atctaccatg gactacaaag acgatgacga caagcttgcg gccgcgaatt caagcttagc 1441 caccatggac tacaaagacg atgacgataa agcaaggctc gaatcggtac ctaaggatcc 1501 cgggcagcag ccgccgcctc aaccggcccc ccagggccaa gggcagccgc cttcgcagcc 1561 cccgcagggg cagggcccgc cgtccggacc cgggcaaccg gcacccgcgg cgacccaggc 1621 ggcgccgcag gcaccccccg ccgggcatca gatcgtgcac gtccgcgggg acgcggagac 1681 cgacctggag gcgctcttca acgccgtcat gaaccccaag acggccaacg tgccccagac 1741 cgtgcccatg aggctccgga agctgcccga cgccttcttc aagccgccgg agcccaaatc 1801 ccactcccga caggccgcta ctgatgcagg cactgcagga gccctgactc cacagcatgt 1861 tcgagctcat gccgctccag ctgctctgca gttgggagct gtttctcctg ggacactgac 1921 ccccactgga gtagtctctg gcccagcagc tacacccaca gctcagcatc ttcgacaggc 1981 tgcttttgag atacctgatg atgtacctct gccagcaggt tgggagatgg caaagacatc 2041 ttctggtcag agatacttct taaatcacat cgatcagaca acaacatggc aggaccccag 2101 gaaggccatg ctgtcccaga tgaacgtcac agcccccacc agtccaccag tgcagcagaa 2161 tatgatgaac tcggcttcag gtcctcttcc tgatggatgg gaacaagcca tgactcagga 2221 tggagaaatt tactatataa accataagaa caagaccacc tcttggctag acccaaggct 2281 tgaccctcgt tttgccatga accagagaat cagtcagagt gctccagtga aacagccacc 2341 acccctggct ccccagagcc cacagggagg cgtcatgggt ggcagcaact ccaaccagca 2401 gcaacagatg cgactgcagc aactgcagat ggagaaggag aggctgcggc tgaaacagca 2461 agaactgctt cggcaggagt tagccctgcg tagccagtta ccaacactgg agcaggatgg 2521 tgggactcaa aatccagtgt cttctcccgg gatgtctcag gaattgagaa caatgacgac 2581 caatagctca gatcctttcc ttaacagtgg cacctatcac tctcgagatg aggctacaga 2641 cagtggacta agcatgagca gctacagtgt ccctcgaacc ccagatgact tcctgaacag 2701 tgtggatgag atggatacag gtgatactat caaccaaagc accctgccct cacagcagaa 2761 ccgtttccca gactaccttg aagccattcc tgggacaaat gtggaccttg gaacactgga 2821 aggagatgga atgaacatag aaggagagga gctgatgcca agtctgcagg aagctttgag 2881 ttctgacatc cttaatgaca tggagtctgt tttggctgcc accaagctag ataaagaaag 2941 ctttcttaca tggttataga gccctcaggc agactgaatt ctgcagatat ccagcacagt 3001 ggcggccgct cgagtctaga ggatcccggg tggcatccct gtgacccctc cccagtgcct 3061 ctcctggccc tggaagttgc cactccagtg cccaccagcc ttgtcctaat aaaattaagt 3121 tgcatcattt tgtctgacta ggtgtccttc tataatatta tggggtggag gggggtggta 3181 tggagcaagg ggcaagttgg gaagacaacc tgtagggcct gcggggtcta ttgggaacca 3241 agctggagtg cagtggcaca atcttggctc actgcaatct ccgcctcctg ggttcaagcg 3301 attctcctgc ctcagcctcc cgagttgttg ggattccagg catgcatgac caggctcagc 3361 taatttttgt ttttttggta gagacggggt ttcaccatat tggccaggct ggtctccaac 3421 tcctaatctc aggtgatcta cccaccttgg cctcccaaat tgctgggatt acaggcgtga 3481 accactgctc ccttccctgt ccttctgatt ttaaaataac tataccagca ggaggacgtc 3541 cagacacagc ataggctacc tggccatgcc caaccggtgg gacatttgag ttgcttgctt 3601 ggcactgtcc tctcatgcgt tgggtccact cagtagatgc ctgttgaatt gggtacgcgg 3661 ccagcttggc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 3721 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 3781 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 3841 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 3901 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 3961 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcctcgag 4021 gaactgaaaa accagaaagt taattcccta tagtgagtcg tattaaattc gtaatcatgt 4081 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg 4141 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt 4201 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg 4261 gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg 4321 actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa 4381 tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc 4441 aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc 4501 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat 4561 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc 4621 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct 4681 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg 4741 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc 4801 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga 4861 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa 4921 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta 4981 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc 5041 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg 5101 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga 5161 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg 5221 agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct 5281 gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg 5341 agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc 5401 cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa 5461 ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc 5521 cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt 5581 cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc 5641 ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt 5701 tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc 5761 catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt 5821 gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata 5881 gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga 5941 tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag 6001 catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa 6061 aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt 6121 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga 6181 aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct g //
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