| 质粒名称: | pT3-EF1aH Yap S127A |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 86497 |
| 存储实验室: | Xin Chen |
|
载体骨架基本信息 |
|
| 载体名称: | pT3-EF1aH |
| 完整质粒大小: | 6693bp |
| 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 |
|
| 插入基因名称: | Yes associated protein 1 / YAP1 isoform 3 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1467 bp |
| 基因突变: | S127A |
| GenBank编号 | NM_0061069 |
| 启动子: | EF1a |
| 标签/融合蛋白: | FLAG (N terminal on backbone) |
| 克隆方法: | Gateway Cloning |
| 5’测序引物 | EF1a-F |
|
文献引用方法 |
|
| 材料和方法部分: | pT3-EF1aH Yap S127A was a gift from Xin Chen (Addgene plasmid # 86497 ; http://n2t.net/addgene:86497 ; RRID:Addgene_86497) |
| 参考文献部分: | Yes-Associated Protein Up-regulates Jagged-1 and Activates the NOTCH Pathway in Human Hepatocellular Carcinoma. Tschaharganeh DF, Chen X, Latzko P, Malz M, Gaida MM, Felix K, Ladu S, Singer S, Pinna F, Gretz N, Sticht C, Tomasi ML, Delogu S, Evert M, Fan B, Ribback S, Jiang L, Brozzetti S, Bergmann F, Dombrowski F, Schirmacher P, Calvisi DF, Breuhahn K. Gastroenterology. 2013 Jun;144(7):1530-1542.e12. doi: 10.1053/j.gastro.2013.02.009. Epub 2013 Feb 16. 10.1053/j.gastro.2013.02.009 PubMed 23419361 |
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 pT3-EF1aH Yap S127A 6693 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian expression of FLAG-tagged Yap containing S127A mutation.
ACCESSION .
VERSION .
KEYWORDS pT3-EF1aH Yap S127A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6693)
AUTHORS Tschaharganeh DF, Chen X, Latzko P, Malz M, Gaida MM, Felix K, Ladu
S, Singer S, Pinna F, Gretz N, Sticht C, Tomasi ML, Delogu S, Evert
M, Fan B, Ribback S, Jiang L, Brozzetti S, Bergmann F, Dombrowski F,
Schirmacher P, Calvisi DF, Breuhahn K
TITLE Yes-Associated Protein Up-regulates Jagged-1 and Activates the NOTCH
Pathway in Human Hepatocellular Carcinoma.
JOURNAL Gastroenterology. 2013 Jun;144(7):1530-1542.e12. doi:
10.1053/j.gastro.2013.02.009. Epub 2013 Feb 16.
PUBMED 23419361
REFERENCE 2 (bases 1 to 6693)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..6693
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..1179
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 232..1170
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 1127..1147
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
protein_bind 1210..1234
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1265..1288
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
protein_bind complement(2800..2824)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
primer_bind 2852..2871
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(2906..2923)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2912..3136
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(3554..3570)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3554..3570)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3567..3589)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3578..3594
/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(3602..3632)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3647..3668
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3785..3802)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3956..4544)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4036..4055)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4715..5575)
/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 5338..5357
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5576..5680)
/gene="bla"
/label=AmpR promoter
primer_bind 5748..5766
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(5804..5826)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 5926..5945
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 6139..6161
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6153..6170
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6154..6170
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6552..6572)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
1 ggctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
61 ggaggggtcg gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
121 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
181 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc
241 gtgtgtggtt cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt
301 acttccacct ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg
361 gagagttcga ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg
421 cctgggcgct ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct
481 ttcgataagt ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg
541 caagatagtc ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc
601 gcgggcggcg acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga
661 gcgcggccac cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct
721 ggcctcgcgc cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca
781 gttgcgtgag cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg
841 acgcggcgct cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg
901 tcctcagccg tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat
961 tagttctcga gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg
1021 gagtttcccc acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa
1081 ttctccttgg aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca
1141 gtggttcaaa gtttttttct tccatttcag gtgtcgtgag gaattagctt ggtaccaata
1201 cggatatcaa caagtttgta caaaaaagca ggctttaaag gaaccaattc agtcgaccac
1261 catggactac aaagacgatg acgataaagc aaggctcgaa tcggtaccta tggatcccgg
1321 gcagcagccg ccgcctcaac cggcccccca gggccaaggg cagccgcctt cgcagccccc
1381 gcaggggcag ggcccgccgt ccggacccgg gcaaccggca cccgcggcga cccaggcggc
1441 gccgcaggca ccccccgccg ggcatcagat cgtgcacgtc cgcggggact cggagaccga
1501 cctggaggcg ctcttcaacg ccgtcatgaa ccccaagacg gccaacgtgc cccagaccgt
1561 gcccatgagg ctccggaagc tgcccgactc cttcttcaag ccgccggagc ccaaatccca
1621 ctcccgacag gccagtactg atgcaggcac tgcaggagcc ctgactccac agcatgttcg
1681 agctcatgcc tctccagctt ctctgcagtt gggagctgtt tctcctggga cactgacccc
1741 cactggagta gtctctggcc cagcagctac acccacagct cagcatcttc gacagtcttc
1801 ttttgagata cctgatgatg tacctctgcc agcaggttgg gagatggcaa agacatcttc
1861 tggtcagaga tacttcttaa atcacatcga tcagacaaca acatggcagg accccaggaa
1921 ggccatgctg tcccagatga acgtcacagc ccccaccagt ccaccagtgc agcagaatat
1981 gatgaactcg gcttcaggtc ctcttcctga tggatgggaa caagccatga ctcaggatgg
2041 agaaatttac tatataaacc ataagaacaa gaccacctct tggctagacc caaggcttga
2101 ccctcgtttt gccatgaacc agagaatcag tcagagtgct ccagtgaaac agccaccacc
2161 cctggctccc cagagcccac agggaggcgt catgggtggc agcaactcca accagcagca
2221 acagatgcga ctgcagcaac tgcagatgga gaaggagagg ctgcggctga aacagcaaga
2281 actgcttcgg caggagttag ccctgcgtag ccagttacca acactggagc aggatggtgg
2341 gactcaaaat ccagtgtctt ctcccgggat gtctcaggaa ttgagaacaa tgacgaccaa
2401 tagctcagat cctttcctta acagtggcac ctatcactct cgagatgaga gtacagacag
2461 tggactaagc atgagcagct acagtgtccc tcgaacccca gatgacttcc tgaacagtgt
2521 ggatgagatg gatacaggtg atactatcaa ccaaagcacc ctgccctcac agcagaaccg
2581 tttcccagac taccttgaag ccattcctgg gacaaatgtg gaccttggaa cactggaagg
2641 agatggaatg aacatagaag gagaggagct gatgccaagt ctgcaggaag ctttgagttc
2701 tgacatcctt aatgacatgg agtctgtttt ggctgccacc aagctagata aagaaagctt
2761 tcttacatgg ttataggcgg ccgcactcga gatatctaga cccagctttc ttgtacaaag
2821 tggttgatat cgcggccgct gcattctagt tgtggtttgt ccaaactcat caatgtatct
2881 tatcatgtct ggatccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
2941 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
3001 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
3061 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
3121 gcggtgggct ctatggcttc tgaggcggga tccccttgaa atacatccac aggtacacct
3181 ccaattgact caaatgatgt caattagtct atcagaagct tctaaagcca tgacatcatt
3241 ttctggaatt ttccaagctg tttaaaggca cagtcaactt agtgtatgta aacttctgac
3301 ccactggaat tgtgatacag tgaattataa gtgaaataat ctgtctgtaa acaattgttg
3361 gaaaaatgac ttgtgtcatg cacaaagtag atgtcctaac tgacttgcca aaactattgt
3421 ttgttaacaa gaaatttgtg gagtagttga aaaacgagtt ttaatgactc caacttaagt
3481 gtatgtaaac ttccgacttc aactgtatag gtctagagtc gacctgcagg catgcaagct
3541 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
3601 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
3661 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
3721 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
3781 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
3841 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
3901 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
3961 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
4021 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
4081 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
4141 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
4201 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
4261 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
4321 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
4381 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
4441 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
4501 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
4561 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
4621 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
4681 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
4741 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
4801 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
4861 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
4921 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
4981 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
5041 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
5101 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
5161 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
5221 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
5281 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
5341 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
5401 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
5461 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
5521 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
5581 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
5641 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
5701 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
5761 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
5821 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
5881 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
5941 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
6001 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
6061 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
6121 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggccagt gaattcgagc
6181 tcggtaccct acagttgaag tcggaagttt acatacactt aagttggagt cattaaaact
6241 cgtttttcaa ctactccaca aatttcttgt taacaaacaa tagttttggc aagtcagtta
6301 ggacatctac tttgtgcatg acacaagtca tttttccaac aattgtttac agacagatta
6361 tttcacttat aattcactgt atcacaattc cagtgggtca gaagtttaca tacactaagt
6421 tgactgtgcc tttaaacagc ttggaaaatt ccagaaaatg atgtcatggc tttagaagct
6481 tctgatagac taattgacat catttgagtc aattggaggt gtacctgtgg atgtatttca
6541 aggaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
6601 cagaagtatg caaagcatgc atatcgatac tagtttaatt aagatcctct ttgcagctaa
6661 tggaccttct aggtcttgaa aggagtggga att
//
