| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-shRNA1 , pLVX shRNA1 |
| 质粒类型: | 哺乳动物细胞慢病毒表达载体，shRNA，RNA干扰载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 启动子: | CMV |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 7758 bp |
| 5' 测序引物及序列: | LKO.1 5’：GACTATCATATGCTTACCGT |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 氨苄 |
| 筛选标记: | 嘌呤霉素 |
| 备注: | 含有组成型人U6启动子的慢病毒shRNA载体 |
| 产品目录号: | 632177 |
| 稳定性: | / |
| 组成型: | -- |
| 病毒/非病毒: | 慢病毒 |
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
Description
pLVX-shRNA1 is an HIV-1-based, lentiviral expression vector designed to express a small hair-pin RNA (shRNA) for RNA interference (RNAi) studies. Expression of your shRNA is drivenby the RNA Pol III-dependent, human U6 promoter (PU6), located just upstream of the MCS. pLVX-shRNA1 can either be used as a plasmid expression vector and transfected into cells, or it can be packaged into viral particles and transduced into cells. Lentiviral particles derived from the vector allow the expression of shRNAs in virtually any cell type, including primary cells.
pLVX-shRNA1 contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA (1), leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-shRNA1 also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, thiselement creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (3).
In addition to lentiviral elements, pLVX-shRNA1 contains a puromycin resistance gene (Puror)under the control of the murine phosphoglycerate kinase promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.
Use
pLVX-shRNA1 is available in the Lenti-X™ shRNA Expression System (Cat. No. 632177). Before it can be transduced into target cells, the vector must be cotransfected into 293T cells with our Lenti-X HTX Packaging System (Cat. Nos. 631247 and 631249) and packaged into viral particles. This packaging system allows the safe production of high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (4).
载体序列
LOCUS pLVX-shRNA1 7757 bp DNA circular 8-NOV-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 632177,632178
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
promoter 2457..2965
/label=PGK\promoter
CDS 2986..3585
/label=Puro(R)
misc_feature 2187..2444
/label=Human\U6\promoter
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=packaging\signal
misc_feature 2028..2151
/label=cPPT/CTS\(DNA\flap)
misc_feature 3679..4270
/label=WPRE
rep_origin complement(5578..6251)
/label=pUC\origin
CDS complement(6396..7392)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
LTR 551..635
/label=U5
misc_binding 636..653
/label=PBS
LTR 4473..5109
/label=3'\LTR
BASE COUNT 1968 a 1930 c 2022 g 1837 t
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
2161 acagcagaga tccagtttat cgatctgggc aggaagaggg cctatttccc atgattcctt
2221 catatttgca tatacgatac aaggctgtta gagagataat tagaattaat ttgactgtaa
2281 acacaaagat attagtacaa aatacgtgac gtagaaagta ataatttctt gggtagtttg
2341 cagttttaaa attatgtttt aaaatggact atcatatgct taccgtaact tgaaagtatt
2401 tcgatttctt ggctttatat atcttgtgga aaggacgagg atccggacaa gcttcgaatt
2461 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
2521 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
2581 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
2641 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
2701 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
2761 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
2821 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
2881 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
2941 ctcttcctca tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag
3001 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
3061 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
3121 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
3181 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
3241 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
3301 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
3361 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
3421 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
3481 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
3541 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgacgccc gccccacgac
3601 ccgcagcgcc cgaccgaaag gagcgcacga ccccatgcat cggcgtctcg agtctagagg
3661 gcccgacgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
3721 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
3781 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
3841 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
3901 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
3961 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
4021 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
4081 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
4141 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
4201 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
4261 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
4321 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
4381 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
4441 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
4501 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
4561 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
4621 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
4681 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
4741 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
4801 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
4861 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
4921 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
4981 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
5041 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
5101 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
5161 atatcagaga gtgagaggcc ttgacattgc tagcgtttac cgtcgacctc tagctagagc
5221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
5281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
5341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
5401 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
5461 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
5521 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
5581 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
5641 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
5701 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
5761 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
5821 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
5881 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
5941 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
6001 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
6061 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
6121 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
6181 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
6241 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
6301 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
6361 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
6421 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
6481 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
6541 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
6601 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
6661 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
6721 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
6781 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
6841 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
6901 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
6961 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
7021 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
7081 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
7141 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
7201 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
7261 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
7321 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
7381 ccacctgacg tcgacggatc gggagatcaa cttgtttatt gcagcttata atggttacaa
7441 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
7501 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atcaactgga taactcaagc
7561 taaccaaaat catcccaaac ttcccacccc ataccctatt accactgcca attacctgtg
7621 gtttcattta ctctaaacct gtgattcctc tgaattattt tcattttaaa gaaattgtat
7681 ttgttaaata tgtactacaa acttagtagt ttttaaagaa attgtatttg ttaaatatgt
7741 actacaaact tagtagt
//
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