| 质粒名称: | pFLAG-YAP1 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66853 |
| 存储实验室: | Yutaka Hata |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV FLAG |
| 载体出品公司: | Kodak (Sigma) |
| 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: | YAP1,Yes-associated protein 1 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | 10413 NM_001130145.2 |
| GenBank编号 | -- |
| 启动子 | CMV |
| 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: | BamHI（克隆过程中销毁） |
| 3' 克隆位点: | KpnI (未破坏) |
| 5' 测序引物 | CMV promoter |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pFLAG-YAP1 was a gift from Yutaka Hata (Addgene plasmid # 66853 ; http://n2t.net/addgene:66853 ; RRID:Addgene_66853) |
| 参考文献部分: | A cell-based assay to screen stimulators of the Hippo pathway reveals the inhibitory effect of dobutamine on the YAP-dependent gene transcription. Bao Y, Nakagawa K, Yang Z, Ikeda M, Withanage K, Ishigami-Yuasa M, Okuno Y, Hata S, Nishina H, Hata Y. J Biochem. 2011 Aug;150(2):199-208. doi: 10.1093/jb/mvr063. Epub 2011 May 17. 10.1093/jb/mvr063 PubMed 21586534 |
买家导航
pFLAGYAP1载体基本信息
pFLAGYAP1载体图谱质粒图谱和多克隆位点信息
pFLAGYAP1载体简介
pFLAGYAP1载体序列
LOCUS pFLAG-YAP1 6212 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express FLAG-tagged YAP1 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pFLAG-YAP1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6212)
AUTHORS Bao Y, Nakagawa K, Yang Z, Ikeda M, Withanage K, Ishigami-Yuasa M,
Okuno Y, Hata S, Nishina H, Hata Y
TITLE A cell-based assay to screen stimulators of the Hippo pathway
reveals the inhibitory effect of dobutamine on the YAP-dependent
gene transcription.
JOURNAL J Biochem. 2011 Aug;150(2):199-208. doi: 10.1093/jb/mvr063. Epub
2011 May 17.
PUBMED 21586534
REFERENCE 2 (bases 1 to 6212)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6212
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 126..148
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 140..157
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 141..157
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
enhancer 318..697
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 698..901
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 852..872
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 935..958
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 2553..3175
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2752..2771)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(3199..3219)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3204..3533
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3384..3519
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3446..3465
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3571..3590)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3572..3590)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3604..3620)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3604..3620)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3617..3639)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3628..3644
/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(3652..3682)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3697..3718
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3835..3852)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4006..4594)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4086..4105)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4765..5625)
/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 5388..5407
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5626..5730)
/gene="bla"
/label=AmpR promoter
rep_origin 5757..6212
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5844..5863)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6054..6075
/label=F1ori-F
/note="F1 origin, forward primer"
ORIGIN
1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
901 tcgtttagtg aaccgtcaga attgatctac catggactac aaagacgatg acgacaagct
961 tgcggccgcg aattcatcga tagatcccgg gcagcagccg ccgcctcaac cggcccccca
1021 gggccaaggg cagccgcctt cgcagccccc gcaggggcag ggcccgccgt ccggacccgg
1081 gcaaccggca cccgcggcga cccaggcggc gccgcaggca ccccccgccg ggcatcagat
1141 cgtgcacgtc cgcggggact cggagaccga cctggaggcg ctcttcaacg ccgtcatgaa
1201 ccccaagacg gccaacgtgc cccagaccgt gcccatgagg ctccggaagc tgcccgactc
1261 cttcttcaag ccgccggagc ccaaatccca ctcccgacag gccagtactg atgcaggcac
1321 tgcaggagcc ctgactccac agcatgttcg agctcattcc tctccagctt ctctgcagtt
1381 gggagctgtt tctcctggga cactgacccc cactggagta gtctctggcc cagcagctac
1441 acccacagct cagcatcttc gacagtcttc ttttgagata cctgatgatg tacctctgcc
1501 agcaggttgg gagatggcaa agacatcttc tggtcagaga tacttcttaa atcacatcga
1561 tcagacaaca acatggcagg accccaggaa ggccatgctg tcccagatga acgtcacagc
1621 ccccaccagt ccaccagtgc agcagaatat gatgaactcg gcttcaggtc ctcttcctga
1681 tggatgggaa caagccatga ctcaggatgg agaaatttac tatataaacc ataagaacaa
1741 gaccacctct tggctagacc caaggcttga ccctcgtttt gccatgaacc agagaatcag
1801 tcagagtgct ccagtgaaac agccaccacc cctggctccc cagagcccac agggaggcgt
1861 catgggtggc agcaactcca accagcagca acagatgcga ctgcagcaac tgcagatgga
1921 gaaggagagg ctgcggctga aacagcaaga actgcttcgg caggcaatgc ggaatatcaa
1981 tcccagcaca gcaaattctc caaaatgtca ggagttagcc ctgcgtagcc agttaccaac
2041 actggagcag gatggtggga ctcaaaatcc agtgtcttct cccgggatgt ctcaggaatt
2101 gagaacaatg acgaccaata gctcagatcc tttccttaac agtggcacct atcactctcg
2161 agatgagagt acagacagtg gactaagcat gagcagctac agtgtccctc gaaccccaga
2221 tgacttcctg aacagtgtgg atgagatgga tacaggtgat actatcaacc aaagcaccct
2281 gccctcacag cagaaccgtt tcccagacta ccttgaagcc attcctggga caaatgtgga
2341 ccttggaaca ctggaaggag atggaatgaa catagaagga gaggagctga tgccaagtct
2401 gcaggaagct ttgagttctg acatccttaa tgacatggag tctgttttgg ctgccaccaa
2461 gctagataaa gaaagctttc ttacatggtt atagagccct caggcagact gaattctgca
2521 gtcgacggta ccagtcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag
2581 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
2641 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
2701 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
2761 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
2821 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
2881 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
2941 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
3001 cgtgaaccac tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg
3061 acgtccagac acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct
3121 tgcttggcac tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta
3181 cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
3241 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
3301 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
3361 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
3421 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
3481 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
3541 tcgaggaact gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat
3601 catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
3661 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
3721 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
3781 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
3841 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
3901 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
3961 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
4021 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
4081 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
4141 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
4201 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
4261 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
4321 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
4381 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
4441 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
4501 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
4561 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
4621 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
4681 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
4741 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
4801 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
4861 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
4921 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
4981 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
5041 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
5101 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
5161 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
5221 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
5281 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
5341 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
5401 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
5461 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
5521 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
5581 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
5641 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
5701 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc
5761 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
5821 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
5881 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
5941 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
6001 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
6061 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
6121 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
6181 gaattttaac aaaatattaa cgcttacaat tt
//
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