| 质粒名称: | shp53 pLKO.1 puro |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 19119 |
| 存储实验室: | Bob Weinberg |
|
载体骨架基本信息 |
|
| 载体名称: | pLKO.1 puro |
| 载体出品公司: | GenHunter |
| 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: | small hairpin RNA against tumor protein p53,p53, FLJ92943, LFS1, TRP53 |
| 物种来源: | H. sapiens (human) |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 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: | shp53 pLKO.1 puro was a gift from Bob Weinberg (Addgene plasmid # 19119 ; http://n2t.net/addgene:19119 ; RRID:Addgene_19119) |
| 参考文献部分: | Growth-inhibitory and tumor- suppressive functions of p53 depend on its repression of CD44 expression. Godar S, Ince TA, Bell GW, Feldser D, Donaher JL, Bergh J, Liu A, Miu K, Watnick RS, Reinhardt F, McAllister SS, Jacks T, Weinberg RA. Cell. 2008 Jul 11. 134(1):62-73. 10.1016/j.cell.2008.06.006 PubMed 18614011 |
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 shp53 pLKO.1 puro 7081 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION 3rd gen lentiviral vector for knocking down p53 gene expression.
ACCESSION .
VERSION .
KEYWORDS shp53 pLKO.1 puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7081)
AUTHORS Godar S, Ince TA, Bell GW, Feldser D, Donaher JL, Bergh J, Liu A,
Miu K, Watnick RS, Reinhardt F, McAllister SS, Jacks T, Weinberg RA
TITLE Growth-inhibitory and tumor- suppressive functions of p53 depend on
its repression of CD44 expression.
JOURNAL Cell. 2008 Jul 11. 134(1):62-73.
PUBMED 18614011
REFERENCE 2 (bases 1 to 7081)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..7081
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind 107..128
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 186..208
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 240..260
/label=T3
/note="T3 promoter, forward primer"
promoter 242..260
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 288..514
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 515..695
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 742..867
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1360..1593
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
promoter 2120..2360
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 2120..2140
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2291..2310
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_feature 2462..2579
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2628..3138
/label=hPGK promoter
/note="human phosphoglycerate kinase 1 promoter"
primer_bind 3057..3075
/label=hPGK-F
/note="Human PGK promoter, forward primer"
CDS 3160..3759
/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(3160..3179)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 3656..3676
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 3887..4120
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 4192..4313
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4229..4248)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4283..4302
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 4353..4488
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 4415..4434
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(4508..4527)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(4509..4527)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4537..4554)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(4537..4553)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4546..4568)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 4695..5150
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(4782..4801)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 4992..5013
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 5176..5280
/gene="bla"
/label=AmpR promoter
CDS 5281..6141
/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 complement(5499..5518)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 6312..6900
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 6801..6820
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7054..7071
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg
241 caattaaccc tcactaaagg gaacaaaagc tggagctgca agcttaatgt agtcttatgc
301 aatactcttg tagtcttgca acatggtaac gatgagttag caacatgcct tacaaggaga
361 gaaaaagcac cgtgcatgcc gattggtgga agtaaggtgg tacgatcgtg ccttattagg
421 aaggcaacag acgggtctga catggattgg acgaaccact gaattgccgc attgcagaga
481 tattgtattt aagtgcctag ctcgatacat aaacgggtct ctctggttag accagatctg
541 agcctgggag ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc
601 ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct
661 cagacccttt tagtcagtgt ggaaaatctc tagcagtggc gcccgaacag ggacttgaaa
721 gcgaaaggga aaccagagga gctctctcga cgcaggactc ggcttgctga agcgcgcacg
781 gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag cggaggctag
841 aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag atcgcgatgg
901 gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca tatagtatgg
961 gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac atcagaaggc
1021 tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga agaacttaga
1081 tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga gataaaagac
1141 accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac caccgcacag
1201 caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt ggagaagtga
1261 attatataaa tataaagtag taaaaattga accattagga gtagcaccca ccaaggcaaa
1321 gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt tccttgggtt
1381 cttgggagca gcaggaagca ctatgggcgc agcgtcaatg acgctgacgg tacaggccag
1441 acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta ttgaggcgca
1501 acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa gaatcctggc
1561 tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct ctggaaaact
1621 catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc tggaacagat
1681 ttggaatcac acgacctgga tggagtggga cagagaaatt aacaattaca caagcttaat
1741 acactcctta attgaagaat cgcaaaacca gcaagaaaag aatgaacaag aattattgga
1801 attagataaa tgggcaagtt tgtggaattg gtttaacata acaaattggc tgtggtatat
1861 aaaattattc ataatgatag taggaggctt ggtaggttta agaatagttt ttgctgtact
1921 ttctatagtg aatagagtta ggcagggata ttcaccatta tcgtttcaga cccacctccc
1981 aaccccgagg ggacccgaca ggcccgaagg aatagaagaa gaaggtggag agagagacag
2041 agacagatcc attcgattag tgaacggatc tcgacggtat cgatcacgag actagcctcg
2101 agcggccgcc cccttcaccg agggcctatt tcccatgatt ccttcatatt tgcatatacg
2161 atacaaggct gttagagaga taattggaat taatttgact gtaaacacaa agatattagt
2221 acaaaatacg tgacgtagaa agtaataatt tcttgggtag tttgcagttt taaaattatg
2281 ttttaaaatg gactatcata tgcttaccgt aacttgaaag tatttcgatt tcttggcttt
2341 atatatcttg tggaaaggac gaaacaccga ctccagtggt aatctacttc aagagagtag
2401 attaccactg gagtcttttt gaattctcga cctcgagaca aatggcagta ttcatccaca
2461 attttaaaag aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa
2521 tagcaacaga catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaattttc
2581 gggtttatta cagggacagc agagatccac tttggccgcg gctcgagggg gttggggttg
2641 cgccttttcc aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc
2701 gggaaacgca gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc
2761 acccggatct tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta
2821 agtcgggaag gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt
2881 ctcactagta ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg ccgaccgcga
2941 tgggctgtgg ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg
3001 gcggtgcggg aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt
3061 tccgcattct gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc
3121 accgacctct ctccccaggg ggatccaccg gagcttacca tgaccgagta caagcccacg
3181 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
3241 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
3301 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
3361 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
3421 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
3481 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
3541 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
3601 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
3661 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
3721 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac gcccgcccca cgacccgcag
3781 cgcccgaccg aaaggagcgc acgaccccat gcatcggtac ctttaagacc aatgacttac
3841 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
3901 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
3961 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
4021 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
4081 tccctcagac ccttttagtc agtgtggaaa atctctagca gtagtagttc atgtcatctt
4141 attattcagt atttataact tgcaaagaaa tgaatatcag agagtgagag gaacttgttt
4201 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
4261 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
4321 tggctctagc tatcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
4381 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
4441 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta gggacgtacc
4501 caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt tacaacgtcg
4561 tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc
4621 cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct
4681 gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
4741 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
4801 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
4861 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
4921 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
4981 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
5041 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
5101 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
5161 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
5221 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
5281 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
5341 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
5401 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
5461 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
5521 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
5581 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
5641 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
5701 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
5761 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
5821 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
5881 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
5941 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
6001 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
6061 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
6121 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
6181 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
6241 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
6301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
6361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
6421 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
6481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
6541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
6601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
6661 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
6721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
6781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
6841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
6901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
6961 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
7021 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
7081 g
//
