| 质粒名称: | shYAP1 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 42540 |
| 存储实验室: | William Hahn |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1 |
| 载体出品公司: | TRC |
| 空载体大小: | 8000 bp |
| 完整质粒大小: | 7084 bp |
| 载体类型: | 哺乳细胞表达、Lentiviral, RNAi |
| 载体标识： | Puromycin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | YAP1 |
| 物种来源: | H. sapiens (human) |
| GenBank编号 | NM_001130145 |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | Age1 (destroyed during cloning) |
| 3’克隆位点: | EcoR1 (destroyed during cloning) |
| 5’测序引物 | pLKOF |
| 3’测序引物 | pLKOR |
|
文献引用方法 |
|
| 材料和方法部分: |
shYAP1 # 1 was a gift from William Hahn (Addgene plasmid # 42540 ; http://n2t.net/addgene:42540 ; RRID:Addgene_42540)
|
| 参考文献部分: | beta-Catenin-Driven Cancers Require a YAP1 Transcriptional Complex for Survival and Tumorigenesis. Rosenbluh J, Nijhawan D, Cox AG, Li X, Neal JT, Schafer EJ, Zack TI, Wang X, Tsherniak A, Schinzel AC, Shao DD, Schumacher SE, Weir BA, Vazquez F, Cowley GS, Root DE, Mesirov JP, Beroukhim R, Kuo CJ, Goessling W, Hahn WC. Cell. 2012 Dec 21;151(7):1457-73. doi: 10.1016/j.cell.2012.11.026. Epub 2012 Dec 13. 10.1016/j.cell.2012.11.026 PubMed 23245941 |
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 shYAP1 # 1 7084 bp ds-DNA circular SYN 23-SEP-2022
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS shYAP1 # 1
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7084)
AUTHORS Rosenbluh J, Nijhawan D, Cox AG, Li X, Neal JT, Schafer EJ, Zack TI,
Wang X, Tsherniak A, Schinzel AC, Shao DD, Schumacher SE, Weir BA,
Vazquez F, Cowley GS, Root DE, Mesirov JP, Beroukhim R, Kuo CJ,
Goessling W, Hahn WC
TITLE beta-Catenin-Driven Cancers Require a YAP1 Transcriptional Complex
for Survival and Tumorigenesis.
JOURNAL Cell. 2012 Dec 21;151(7):1457-73. doi: 10.1016/j.cell.2012.11.026.
Epub 2012 Dec 13.
PUBMED 23245941
REFERENCE 2 (bases 1 to 7084)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 23, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..7084
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(11..28)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(182..770)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(262..281)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(941..1801)
/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 1564..1583
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1802..1906)
/gene="bla"
/label=AmpR promoter
rep_origin complement(1932..2387)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2069..2090)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 2281..2300
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 2514..2536
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2528..2545
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2529..2545
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 2555..2574
/label=T7
/note="T7 promoter, forward primer"
promoter 2555..2573
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 2594..2729
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(2648..2667)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 2769..2890
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(2780..2799)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 2834..2853
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
LTR 2962..3195
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS complement(3323..3922)
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(3406..3426)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 3903..3922
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(3944..4454)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind complement(4007..4025)
/label=hPGK-F
/note="Human PGK promoter, forward primer"
misc_feature 4503..4620
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter complement(4725..4965)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(4775..4794)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(4945..4965)
/label=hU6-F
/note="Human U6 promoter, forward primer"
CDS complement(5263..5307)
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 5492..5725
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 6218..6343
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 6390..6570
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(6571..6797)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(6825..6845)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(6825..6843)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6864..6880)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6864..6880)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6877..6899)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6888..6904
/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(6912..6942)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6957..6978
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
61 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
121 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
181 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
241 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
301 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
361 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
421 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
481 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
541 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
601 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
661 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
721 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
781 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
841 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
901 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
961 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
1021 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
1081 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
1141 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
1201 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
1261 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
1321 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
1381 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
1441 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
1501 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
1561 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
1621 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
1681 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
1741 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
1801 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
1861 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
1921 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
1981 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
2041 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
2101 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
2161 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
2221 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
2281 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
2341 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt
2401 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
2461 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
2521 acgacgttgt aaaacgacgg ccagtgagcg cgcgtaatac gactcactat agggcgaatt
2581 gggtacgtcc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc
2641 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
2701 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggat agctagagcc
2761 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
2821 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
2881 taaacaagtt cctctcactc tctgatattc atttctttgc aagttataaa tactgaataa
2941 taagatgaca tgaactacta ctgctagaga ttttccacac tgactaaaag ggtctgaggg
3001 atctctagtt accagagtca cacaacagac gggcacacac tacttgaagc actcaaggca
3061 agctttattg aggcttaagc agtgggttcc ctagttagcc agagagctcc caggctcaga
3121 tctggtctaa ccagagagac ccagtacaag caaaaagcag atcttgtctt cgttgggagt
3181 gaattagccc ttccagtccc cccttttctt ttaaaaagtg gctaagatct acagctgcct
3241 tgtaagtcat tggtcttaaa ggtaccgatg catggggtcg tgcgctcctt tcggtcgggc
3301 gctgcgggtc gtggggcggg cgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
3361 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
3421 ggttgcgggg cgcggaggtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
3481 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga
3541 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
3601 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
3661 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
3721 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
3781 cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg
3841 cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca
3901 ccgtgggctt gtactcggtc atggtaagct ccggtggatc cccctgggga gagaggtcgg
3961 tgattcggtc aacgagggag ccgactgccg acgtgcgctc cggaggcttg cagaatgcgg
4021 aacaccgcgc gggcaggaac agggcccaca ctaccgcccc acaccccgcc tcccgcaccg
4081 ccccttcccg gccgctgctc tcggcgcgcc ctgctgagca gccgctattg gccacagccc
4141 atcgcggtcg gcgcgctgcc attgctccct ggcgctgtcc gtctgcgagg gtactagtga
4201 gacgtgcggc ttccgtttgt cacgtccggc acgccgcgaa ccgcaaggaa ccttcccgac
4261 ttaggggcgg agcaggaagc gtcgccgggg ggcccacaag ggtagcggcg aagatccggg
4321 tgacgctgcg aacggacgtg aagaatgtgc gagacccagg gtcggcgccg ctgcgtttcc
4381 cggaaccacg cccagagcag ccgcgtccct gcgcaaaccc agggctgcct tggaaaaggc
4441 gcaaccccaa ccccctcgag ccgcggccaa agtggatctc tgctgtccct gtaataaacc
4501 cgaaaatttt gaatttttgt aatttgtttt tgtaattctt tagtttgtat gtctgttgct
4561 attatgtcta ctattctttc ccctgcactg taccccccaa tccccccttt tcttttaaaa
4621 ttgtggatga atactgccat ttgtctcgag gtcgagaatt caaaaagcca ccaagctaga
4681 taaagaactc gagttcttta tctagcttgg tggcccggtg tttcgtcctt tccacaagat
4741 atataaagcc aagaaatcga aatactttca agttacggta agcatatgat agtccatttt
4801 aaaacataat tttaaaactg caaactaccc aagaaattat tactttctac gtcacgtatt
4861 ttgtactaat atctttgtgt ttacagtcaa attaattcca attatctctc taacagcctt
4921 gtatcgtata tgcaaatatg aaggaatcat gggaaatagg ccctcggtga agggggcggc
4981 cgctcgaggc tagtctcgtg atcgataccg tcgagatccg ttcactaatc gaatggatct
5041 gtctctgtct ctctctccac cttcttcttc tattccttcg ggcctgtcgg gtcccctcgg
5101 ggttgggagg tgggtctgaa acgataatgg tgaatatccc tgcctaactc tattcactat
5161 agaaagtaca gcaaaaacta ttcttaaacc taccaagcct cctactatca ttatgaataa
5221 ttttatatac cacagccaat ttgttatgtt aaaccaattc cacaaacttg cccatttatc
5281 taattccaat aattcttgtt cattcttttc ttgctggttt tgcgattctt caattaagga
5341 gtgtattaag cttgtgtaat tgttaatttc tctgtcccac tccatccagg tcgtgtgatt
5401 ccaaatctgt tccagagatt tattactcca actagcattc caaggcacag cagtggtgca
5461 aatgagtttt ccagagcaac cccaaatccc caggagctgt tgatccttta ggtatctttc
5521 cacagccagg attcttgcct ggagctgctt gatgccccag actgtgagtt gcaacagatg
5581 ctgttgcgcc tcaatagccc tcagcaaatt gttctgctgc tgcactatac cagacaataa
5641 ttgtctggcc tgtaccgtca gcgtcattga cgctgcgccc atagtgcttc ctgctgctcc
5701 caagaaccca aggaacaaag ctcctattcc cactgctctt ttttctctct gcaccactct
5761 tctctttgcc ttggtgggtg ctactcctaa tggttcaatt tttactactt tatatttata
5821 taattcactt ctccaattgt ccctcatatc tcctcctcca ggtctgaaga tcagcggccg
5881 cttgctgtgc ggtggtctta cttttgtttt gctcttcctc tatcttgtct aaagcttcct
5941 tggtgtcttt tatctctatc ctttgatgca cacaatagag ggttgctact gtattatata
6001 atgatctaag ttcttctgat cctgtctgaa gggatggttg tagctgtccc agtatttgtc
6061 tacagccttc tgatgtttct aacaggccag gattaactgc gaatcgttct agctccctgc
6121 ttgcccatac tatatgtttt aatttatatt ttttctttcc ccctggcctt aaccgaattt
6181 tttcccatcg cgatctaatt ctcccccgct taatactgac gctctcgcac ccatctctct
6241 ccttctagcc tccgctagtc aaaatttttg gcgtactcac cagtcgccgc ccctcgcctc
6301 ttgccgtgcg cgcttcagca agccgagtcc tgcgtcgaga gagctcctct ggtttccctt
6361 tcgctttcaa gtccctgttc gggcgccact gctagagatt ttccacactg actaaaaggg
6421 tctgagggat ctctagttac cagagtcaca caacagacgg gcacacacta cttgaagcac
6481 tcaaggcaag ctttattgag gcttaagcag tgggttccct agttagccag agagctccca
6541 ggctcagatc tggtctaacc agagagaccc gtttatgtat cgagctaggc acttaaatac
6601 aatatctctg caatgcggca attcagtggt tcgtccaatc catgtcagac ccgtctgttg
6661 ccttcctaat aaggcacgat cgtaccacct tacttccacc aatcggcatg cacggtgctt
6721 tttctctcct tgtaaggcat gttgctaact catcgttacc atgttgcaag actacaagag
6781 tattgcataa gactacatta agcttgcagc tccagctttt gttcccttta gtgagggtta
6841 attgcgcgct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
6901 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
6961 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
7021 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
7081 cgct
//
