| 质粒名称: | p2xFlag CMV2-YAP2-S127A mutant |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 19050 |
| 存储实验室: | Marius Sudol |
|
载体骨架基本信息 |
|
| 载体名称: | p2xFlag CMV2 |
| 空载体大小: | 4700 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: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Yes-kinase associated protein / YAP2, YAP65 / YAP2 isoform |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 1467bp |
| 基因突变: | Serine 127 changed to Alanine |
| GenBank编号 | NM_006106 |
| 标签/融合蛋白: | 2xFlag (N terminal on backbone) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | KpnI(未破坏) |
| 3’克隆位点: | XbaI (未破坏) |
| 5’测序引物 | CMV |
|
文献引用方法 |
|
| 材料和方法部分: |
p2xFlag CMV2-YAP2-S127A mutant was a gift from Marius Sudol (Addgene plasmid # 19050 ; http://n2t.net/addgene:19050 ; RRID:Addgene_19050)
|
| 参考文献部分: | Mst2 and Lats kinases regulate apoptotic function of YAP. Oka T, Mazack V, Sudol M. J Biol Chem. 2008 Jul 17. ():. 10.1074/jbc.M804380200 PubMed 18640976 |
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p2xFlag CMV2-YAP2-S127A mutant载体质粒基本信息
p2xFlag CMV2-YAP2-S127A mutant质粒图谱载体图谱和p2xFlag CMV2-YAP2-S127A mutant载体序列质粒序列多克隆位点信息

p2xFlag CMV2-YAP2-S127A mutant质粒载体简介
p2xFlag CMV2-YAP2-S127A mutant质粒序列载体序列
LOCUS p2xFlag CMV2-YAP2-S127A mutant 6231 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS p2xFlag CMV2-YAP2-S127A mutant
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6231)
AUTHORS Oka T, Mazack V, Sudol M
TITLE Mst2 and Lats kinases regulate apoptotic function of YAP.
JOURNAL J Biol Chem. 2008 Jul 17. ():.
PUBMED 18640976
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"
enhancer 33..412
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 413..616
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 567..587
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
CDS 650..673
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
regulatory 698..707
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 707..730
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
polyA_signal 2287..2909
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
primer_bind complement(2486..2505)
/label=hGH-PA-R
/note="Human growth hormone terminator, reverse primer"
primer_bind complement(2933..2953)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 2938..3267
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3118..3253
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3180..3199
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(3305..3324)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(3306..3324)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(3338..3354)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3338..3354)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3351..3373)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3362..3378
/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(3386..3416)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3431..3452
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(3569..3586)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(3740..4328)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(3820..3839)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4499..5359)
/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 5122..5141
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5360..5464)
/gene="bla"
/label=AmpR promoter
rep_origin 5491..5946
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5578..5597)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5788..5809
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6072..6094
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6086..6103
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6087..6103
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
ORIGIN
1 tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta gttattaata
61 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
121 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
181 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
241 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtccgccccc
301 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttacg
361 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
421 gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
481 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
541 atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac ggtgggaggt
601 ctatataagc agagctcgtt tagtgaaccg tcagaattga tctaccatgg actacaaaga
661 cgatgacgac aagcttgcgg ccgcgaattc aagcttagcc accatggact acaaagacga
721 tgacgataaa gcaaggctcg aatcggtacc tatggatccc gggcagcagc cgccgcctca
781 accggccccc cagggccaag ggcagccgcc ttcgcagccc ccgcaggggc agggcccgcc
841 gtccggaccc gggcaaccgg cacccgcggc gacccaggcg gcgccgcagg caccccccgc
901 cgggcatcag atcgtgcacg tccgcgggga ctcggagacc gacctggagg cgctcttcaa
961 cgccgtcatg aaccccaaga cggccaacgt gccccagacc gtgcccatga ggctccggaa
1021 gctgcccgac tccttcttca agccgccgga gcccaaatcc cactcccgac aggccagtac
1081 tgatgcaggc actgcaggag ccctgactcc acagcatgtt cgagctcatg cctctccagc
1141 ttctctgcag ttgggagctg tttctcctgg gacactgacc cccactggag tagtctctgg
1201 cccagcagct acacccacag ctcagcatct tcgacagtct tcttttgaga tacctgatga
1261 tgtacctctg ccagcaggtt gggagatggc aaagacatct tctggtcaga gatacttctt
1321 aaatcacatc gatcagacaa caacatggca ggaccccagg aaggccatgc tgtcccagat
1381 gaacgtcaca gcccccacca gtccaccagt gcagcagaat atgatgaact cggcttcagg
1441 tcctcttcct gatggatggg aacaagccat gactcaggat ggagaaattt actatataaa
1501 ccataagaac aagaccacct cttggctaga cccaaggctt gaccctcgtt ttgccatgaa
1561 ccagagaatc agtcagagtg ctccagtgaa acagccacca cccctggctc cccagagccc
1621 acagggaggc gtcatgggtg gcagcaactc caaccagcag caacagatgc gactgcagca
1681 actgcagatg gagaaggaga ggctgcggct gaaacagcaa gaactgcttc ggcaggagtt
1741 agccctgcgt agccagttac caacactgga gcaggatggt gggactcaaa atccagtgtc
1801 ttctcccggg atgtctcagg aattgagaac aatgacgacc aatagctcag atcctttcct
1861 taacagtggc acctatcact ctcgagatga gagtacagac agtggactaa gcatgagcag
1921 ctacagtgtc cctcgaaccc cagatgactt cctgaacagt gtggatgaga tggatacagg
1981 tgatactatc aaccaaagca ccctgccctc acagcagaac cgtttcccag actaccttga
2041 agccattcct gggacaaatg tggaccttgg aacactggaa ggagatggaa tgaacataga
2101 aggagaggag ctgatgccaa gtctgcagga agctttgagt tctgacatcc ttaatgacat
2161 ggagtctgtt ttggctgcca ccaagctaga taaagaaagc tttcttacat ggttatagag
2221 ccctcaggca gactgaattc tgcagatatc cagcacagtg gcggccgctc gagtctagag
2281 gatcccgggt ggcatccctg tgacccctcc ccagtgcctc tcctggccct ggaagttgcc
2341 actccagtgc ccaccagcct tgtcctaata aaattaagtt gcatcatttt gtctgactag
2401 gtgtccttct ataatattat ggggtggagg ggggtggtat ggagcaaggg gcaagttggg
2461 aagacaacct gtagggcctg cggggtctat tgggaaccaa gctggagtgc agtggcacaa
2521 tcttggctca ctgcaatctc cgcctcctgg gttcaagcga ttctcctgcc tcagcctccc
2581 gagttgttgg gattccaggc atgcatgacc aggctcagct aatttttgtt tttttggtag
2641 agacggggtt tcaccatatt ggccaggctg gtctccaact cctaatctca ggtgatctac
2701 ccaccttggc ctcccaaatt gctgggatta caggcgtgaa ccactgctcc cttccctgtc
2761 cttctgattt taaaataact ataccagcag gaggacgtcc agacacagca taggctacct
2821 ggccatgccc aaccggtggg acatttgagt tgcttgcttg gcactgtcct ctcatgcgtt
2881 gggtccactc agtagatgcc tgttgaattg ggtacgcggc cagcttggct gtggaatgtg
2941 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
3001 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
3061 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
3121 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
3181 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
3241 ttttttggag gcctaggctt ttgcaaaaag ctcctcgagg aactgaaaaa ccagaaagtt
3301 aattccctat agtgagtcgt attaaattcg taatcatgtc atagctgttt cctgtgtgaa
3361 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
3421 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
3481 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
3541 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
3601 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
3661 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
3721 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
3781 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
3841 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
3901 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
3961 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
4021 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
4081 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
4141 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
4201 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
4261 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
4321 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
4381 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
4441 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
4501 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
4561 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
4621 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
4681 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
4741 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
4801 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
4861 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
4921 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
4981 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
5041 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
5101 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
5161 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
5221 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
5281 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
5341 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
5401 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
5461 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
5521 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
5581 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
5641 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
5701 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
5761 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
5821 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
5881 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
5941 caatttccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc
6001 ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac
6061 gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgccaagc tgatctatac
6121 attgaatcaa tattggcaat tagccatatt agtcattggt tatatagcat aaatcaatat
6181 tggctattgg ccattgcata cgttgtatct atatcataat atgtacattt a
//
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