| 质粒名称: | pLKO-p53-shRNA-941 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 25637 |
| 存储实验室: | Todd Waldman |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1 puro |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7032 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达，慢病毒，RNAi |
| 筛选标记: | 嘌呤霉素 |
| 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: | p53-shRNA-941 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 42 bp |
| 基因突变: | -- |
| GenBank编号 | NM_000546 |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 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 | LKO.1 5' |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pLKO-p53-shRNA-941 was a gift from Todd Waldman (Addgene plasmid # 25637 ; http://n2t.net/addgene:25637 ; RRID:Addgene_25637) |
| 参考文献部分: | Activation of p53-dependent growth suppression in human cells by mutations in PTEN or PIK3CA. Kim JS, Lee C, Bonifant CL, Ressom H, Waldman T. Mol Cell Biol. 2007 Jan . 27(2):662-77. 10.1128/MCB.00537-06 PubMed 17060456 |
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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 pLKO-p53-shRNA-941 7083 bp ds-DNA circular SYN 21-AUG-2019
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pLKO-p53-shRNA-941
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7083)
AUTHORS Kim JS, Lee C, Bonifant CL, Ressom H, Waldman T
TITLE Activation of p53-dependent growth suppression in human cells by
mutations in PTEN or PIK3CA.
JOURNAL Mol Cell Biol. 2007 Jan . 27(2):662-77.
PUBMED 17060456
REFERENCE 2 (bases 1 to 7083)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 21, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7083
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..234
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
CDS complement(362..961)
/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(445..465)
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind 942..961
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
promoter complement(983..1493)
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind complement(1046..1064)
/label=hPGK-F
/note="Human PGK promoter, forward primer"
misc_feature 1542..1659
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter complement(1763..2003)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(1813..1832)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(1983..2003)
/label=hU6-F
/note="Human U6 promoter, forward primer"
misc_feature 2530..2763
/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 3256..3381
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 3428..3608
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(3609..3835)
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
primer_bind complement(3863..3883)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(3863..3881)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(3902..3918)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(3902..3918)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(3915..3937)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 3926..3942
/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(3950..3980)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 3995..4016
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(4133..4150)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4304..4892)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4384..4403)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5063..5923)
/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 5686..5705
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(5924..6028)
/gene="bla"
/label=AmpR promoter
rep_origin complement(6054..6509)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6191..6212)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6403..6422
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6636..6658
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 6650..6667
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 6651..6667
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6677..6696
/label=T7
/note="T7 promoter, forward primer"
promoter 6677..6695
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
rep_origin 6716..6851
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6770..6789)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
polyA_signal 6891..7012
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6902..6921)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 6956..6975
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
ORIGIN
1 tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta ccagagtcac
61 acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga ggcttaagca
121 gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac cagagagacc
181 cagtacaagc aaaaagcaga tcttgtcttc gttgggagtg aattagccct tccagtcccc
241 ccttttcttt taaaaagtgg ctaagatcta cagctgcctt gtaagtcatt ggtcttaaag
301 gtaccgatgc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg tggggcgggc
361 gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc acctcgacgt
421 cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc gcggaggtct
481 ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc acgacggcgc
541 tgcccagacc cttgccctgg tggtcgggcg agacgccgac ggtggccagg aaccacgcgg
601 gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg gccagccggg
661 aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc gcttcgacgc
721 tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac accttgccga
781 tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc tcgatgtggc
841 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
901 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
961 tggtaagctc cggtggatcc ccctggggag agaggtcggt gattcggtca acgagggagc
1021 cgactgccga cgtgcgctcc ggaggcttgc agaatgcgga acaccgcgcg ggcaggaaca
1081 gggcccacac taccgcccca caccccgcct cccgcaccgc cccttcccgg ccgctgctct
1141 cggcgcgccc tgctgagcag ccgctattgg ccacagccca tcgcggtcgg cgcgctgcca
1201 ttgctccctg gcgctgtccg tctgcgaggg tactagtgag acgtgcggct tccgtttgtc
1261 acgtccggca cgccgcgaac cgcaaggaac cttcccgact taggggcgga gcaggaagcg
1321 tcgccggggg gcccacaagg gtagcggcga agatccgggt gacgctgcga acggacgtga
1381 agaatgtgcg agacccaggg tcggcgccgc tgcgtttccc ggaaccacgc ccagagcagc
1441 cgcgtccctg cgcaaaccca gggctgcctt ggaaaaggcg caaccccaac cccctcgagc
1501 cgcggccaaa gtggatctct gctgtccctg taataaaccc gaaaattttg aatttttgta
1561 atttgttttt gtaattcttt agtttgtatg tctgttgcta ttatgtctac tattctttcc
1621 cctgcactgt accccccaat cccccctttt cttttaaaat tgtggatgaa tactgccatt
1681 tgtctcgagg tcgagaatta aaaacaccat ccactacaac tacatctcga gatgtagttg
1741 tagtggatgg tgccggtgtt tcgtcctttc cacaagatat ataaagccaa gaaatcgaaa
1801 tactttcaag ttacggtaag catatgatag tccattttaa aacataattt taaaactgca
1861 aactacccaa gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt
1921 acagtcaaat taattccaat tatctctcta acagccttgt atcgtatatg caaatatgaa
1981 ggaatcatgg gaaataggcc ctcggtgaag ggggcggccg ctcgaggcta gtctcgtgat
2041 cgataccgtc gagatccgtt cactaatcga atggatctgt ctctgtctct ctctccacct
2101 tcttcttcta ttccttcggg cctgtcgggt cccctcgggg ttgggaggtg ggtctgaaac
2161 gataatggtg aatatccctg cctaactcta ttcactatag aaagtacagc aaaaactatt
2221 cttaaaccta ccaagcctcc tactatcatt atgaataatt ttatatacca cagccaattt
2281 gttatgttaa accaattcca caaacttgcc catttatcta attccaataa ttcttgttca
2341 ttcttttctt gctggttttg cgattcttca attaaggagt gtattaagct tgtgtaattg
2401 ttaatttctc tgtcccactc catccaggtc gtgtgattcc aaatctgttc cagagattta
2461 ttactccaac tagcattcca aggcacagca gtggtgcaaa tgagttttcc agagcaaccc
2521 caaatcccca ggagctgttg atcctttagg tatctttcca cagccaggat tcttgcctgg
2581 agctgcttga tgccccagac tgtgagttgc aacagatgct gttgcgcctc aatagccctc
2641 agcaaattgt tctgctgctg cactatacca gacaataatt gtctggcctg taccgtcagc
2701 gtcattgacg ctgcgcccat agtgcttcct gctgctccca agaacccaag gaacaaagct
2761 cctattccca ctgctctttt ttctctctgc accactcttc tctttgcctt ggtgggtgct
2821 actcctaatg gttcaatttt tactacttta tatttatata attcacttct ccaattgtcc
2881 ctcatatctc ctcctccagg tctgaagatc agcggccgct tgctgtgcgg tggtcttact
2941 tttgttttgc tcttcctcta tcttgtctaa agcttccttg gtgtctttta tctctatcct
3001 ttgatgcaca caatagaggg ttgctactgt attatataat gatctaagtt cttctgatcc
3061 tgtctgaagg gatggttgta gctgtcccag tatttgtcta cagccttctg atgtttctaa
3121 caggccagga ttaactgcga atcgttctag ctccctgctt gcccatacta tatgttttaa
3181 tttatatttt ttctttcccc ctggccttaa ccgaattttt tcccatcgcg atctaattct
3241 cccccgctta atactgacgc tctcgcaccc atctctctcc ttctagcctc cgctagtcaa
3301 aatttttggc gtactcacca gtcgccgccc ctcgcctctt gccgtgcgcg cttcagcaag
3361 ccgagtcctg cgtcgagaga gctcctctgg tttccctttc gctttcaagt ccctgttcgg
3421 gcgccactgc tagagatttt ccacactgac taaaagggtc tgagggatct ctagttacca
3481 gagtcacaca acagacgggc acacactact tgaagcactc aaggcaagct ttattgaggc
3541 ttaagcagtg ggttccctag ttagccagag agctcccagg ctcagatctg gtctaaccag
3601 agagacccgt ttatgtatcg agctaggcac ttaaatacaa tatctctgca atgcggcaat
3661 tcagtggttc gtccaatcca tgtcagaccc gtctgttgcc ttcctaataa ggcacgatcg
3721 taccacctta cttccaccaa tcggcatgca cggtgctttt tctctccttg taaggcatgt
3781 tgctaactca tcgttaccat gttgcaagac tacaagagta ttgcataaga ctacattaag
3841 cttgcagctc cagcttttgt tccctttagt gagggttaat tgcgcgcttg gcgtaatcat
3901 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
3961 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
4021 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
4081 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
4141 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
4201 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
4261 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
4321 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
4381 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
4441 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
4501 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
4561 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
4621 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
4681 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
4741 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
4801 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
4861 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
4921 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
4981 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
5041 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
5101 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
5161 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
5221 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
5281 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
5341 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
5401 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
5461 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
5521 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
5581 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
5641 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
5701 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
5761 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
5821 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
5881 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
5941 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
6001 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
6061 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
6121 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
6181 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
6241 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
6301 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
6361 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
6421 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
6481 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc gcaactgttg
6541 ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc
6601 tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac
6661 ggccagtgag cgcgcgtaat acgactcact atagggcgaa ttgggtacgt ccctaggcct
6721 ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc ctcggcctct
6781 gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact gggcggagtt
6841 aggggcggga tgggcggagt taggggcggg atagctagag ccagacatga taagatacat
6901 tgatgagttt ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat
6961 ttgtgatgct attgctttat ttgtaaccat tataagctgc aataaacaag ttcctctcac
7021 tctctgatat tcatttcttt gcaagttata aatactgaat aataagatga catgaactac
7081 tac
//
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