| 质粒名称: | pCMV-flag S127A YAP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 27370 |
| 存储实验室: | Kunliang Guan |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 5000 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | YAP |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1500 bp |
| 基因突变: | S127A ( major phosphorylation site in YAP2) |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | flag（插入N端子） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-flag S127A YAP was a gift from Kunliang Guan (Addgene plasmid # 27370 ; http://n2t.net/addgene:27370 ; RRID:Addgene_27370) |
| 参考文献部分: | Inactivation of YAP oncoprotein by the Hippo pathway is involved in cell contact inhibition and tissue growth control. Zhao B, Wei X, Li W, Udan RS, Yang Q, Kim J, Xie J, Ikenoue T, Yu J, Li L, Zheng P, Ye K, Chinnaiyan A, Halder G, Lai ZC, Guan KL. Genes Dev. 2007 Nov 1. 21(21):2747-61. 10.1101/gad.1602907 PubMed 17974916 |
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 pCMV-flag S127A YAP 6283 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pCMV-flag S127A YAP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6283)
AUTHORS Zhao B, Wei X, Li W, Udan RS, Yang Q, Kim J, Xie J, Ikenoue T, Yu J,
Li L, Zheng P, Ye K, Chinnaiyan A, Halder G, Lai ZC, Guan KL
TITLE Inactivation of YAP oncoprotein by the Hippo pathway is involved in
cell contact inhibition and tissue growth control.
JOURNAL Genes Dev. 2007 Nov 1. 21(21):2747-61.
PUBMED 17974916
REFERENCE 2 (bases 1 to 6283)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6283
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 277..480
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 431..451
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 514..537
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 562..571
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 571..594
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 2151..2773
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2350..2369)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(2797..2817)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2802..3131
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 2982..3117
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3044..3063
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3169..3188)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3170..3188)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3202..3218)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3202..3218)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3215..3237)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3226..3242
/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(3250..3280)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3295..3316
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3433..3450)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3604..4192)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3684..3703)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4363..5223)
/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 4986..5005
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5224..5328)
/gene="bla"
/label=AmpR promoter
rep_origin 5355..5810
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5442..5461)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5652..5673
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 5936..5958
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 5950..5967
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 5951..5967
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata
61 gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc
121 cacttggcag tacatcaagt gtatcatatg ccaagtccgc cccctattga cgtcaatgac
181 ggtaaatggc ccgcctggca ttatgcccag tacatgacct tacgggactt tcctacttgg
241 cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg gcagtacacc
301 aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc
361 aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taataacccc
421 gccccgttga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct
481 cgtttagtga accgtcagaa ttgatctacc atggactaca aagacgatga cgacaagctt
541 gcggccgcga attcaagctt agccaccatg gactacaaag acgatgacga taaagcaagg
601 ctcgaatcgg tacctaagga tcccgggcag cagccgccgc ctcaaccggc cccccagggc
661 caagggcagc cgccttcgca gcccccgcag gggcagggcc cgccgtccgg acccgggcaa
721 ccggcacccg cggcgaccca ggcggcgccg caggcacccc ccgccgggca tcagatcgtg
781 cacgtccgcg gggactcgga gaccgacctg gaggcgctct tcaacgccgt catgaacccc
841 aagacggcca acgtgcccca gaccgtgccc atgaggctcc ggaagctgcc cgactccttc
901 ttcaagccgc cggagcccaa atcccactcc cgacaggcca gtactgatgc aggcactgca
961 ggagccctga ctccacagca tgttcgagct catgcctctc cagcttctct gcagttggga
1021 gctgtttctc ctgggacact gacccccact ggagtagtct ctggcccagc agctacaccc
1081 acagctcagc atcttcgaca gtcttctttt gagatacctg atgatgtacc tctgccagca
1141 ggttgggaga tggcaaagac atcttctggt cagagatact tcttaaatca catcgatcag
1201 acaacaacat ggcaggaccc caggaaggcc atgctgtccc agatgaacgt cacagccccc
1261 accagtccac cagtgcagca gaatatgatg aactcggctt caggtcctct tcctgatgga
1321 tgggaacaag ccatgactca ggatggagaa atttactata taaaccataa gaacaagacc
1381 acctcttggc tagacccaag gcttgaccct cgttttgcca tgaaccagag aatcagtcag
1441 agtgctccag tgaaacagcc accacccctg gctccccaga gcccacaggg aggcgtcatg
1501 ggtggcagca actccaacca gcagcaacag atgcgactgc agcaactgca gatggagaag
1561 gagaggctgc ggctgaaaca gcaagaactg cttcggcagg agttagccct gcgtagccag
1621 ttaccaacac tggagcagga tggtgggact caaaatccag tgtcttctcc cgggatgtct
1681 caggaattga gaacaatgac gaccaatagc tcagatcctt tccttaacag tggcacctat
1741 cactctcgag atgagagtac agacagtgga ctaagcatga gcagctacag tgtccctcga
1801 accccagatg acttcctgaa cagtgtggat gagatggata caggtgatac tatcaaccaa
1861 agcaccctgc cctcacagca gaaccgtttc ccagactacc ttgaagccat tcctgggaca
1921 aatgtggacc ttggaacact ggaaggagat ggaatgaaca tagaaggaga ggagctgatg
1981 ccaagtctgc aggaagcttt gagttctgac atccttaatg acatggagtc tgttttggct
2041 gccaccaagc tagataaaga aagctttctt acatggttat agagccctca ggcagactga
2101 attctgcaga tatccagcac agtggcggcc gctcgagtct agaggatccc gggtggcatc
2161 cctgtgaccc ctccccagtg cctctcctgg ccctggaagt tgccactcca gtgcccacca
2221 gccttgtcct aataaaatta agttgcatca ttttgtctga ctaggtgtcc ttctataata
2281 ttatggggtg gaggggggtg gtatggagca aggggcaagt tgggaagaca acctgtaggg
2341 cctgcggggt ctattgggaa ccaagctgga gtgcagtggc acaatcttgg ctcactgcaa
2401 tctccgcctc ctgggttcaa gcgattctcc tgcctcagcc tcccgagttg ttgggattcc
2461 aggcatgcat gaccaggctc agctaatttt tgtttttttg gtagagacgg ggtttcacca
2521 tattggccag gctggtctcc aactcctaat ctcaggtgat ctacccacct tggcctccca
2581 aattgctggg attacaggcg tgaaccactg ctcccttccc tgtccttctg attttaaaat
2641 aactatacca gcaggaggac gtccagacac agcataggct acctggccat gcccaaccgg
2701 tgggacattt gagttgcttg cttggcactg tcctctcatg cgttgggtcc actcagtaga
2761 tgcctgttga attgggtacg cggccagctt ggctgtggaa tgtgtgtcag ttagggtgtg
2821 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
2881 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
2941 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
3001 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
3061 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
3121 gcttttgcaa aaagctcctc gaggaactga aaaaccagaa agttaattcc ctatagtgag
3181 tcgtattaaa ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac
3241 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
3301 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
3361 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
3421 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
3481 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
3541 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
3601 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
3661 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
3721 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
3781 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
3841 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
3901 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
3961 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
4021 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
4081 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
4141 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
4201 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
4261 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
4321 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
4381 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
4441 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
4501 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
4561 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
4621 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
4681 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
4741 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
4801 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
4861 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
4921 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
4981 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
5041 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
5101 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
5161 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
5221 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
5281 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
5341 aaaagtgcca cctgacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
5401 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
5461 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
5521 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
5581 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
5641 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
5701 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
5761 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt ccattcgcca
5821 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
5881 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
5941 tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa tcaatattgg
6001 caattagcca tattagtcat tggttatata gcataaatca atattggcta ttggccattg
6061 catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc aatatgaccg
6121 ccatgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
6181 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
6241 ccgcccaacg acccccgccc attgacgtca ataatgacgt atg
//
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