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shRNA质粒

pLVX-shRNA1 慢病毒shRNA,RNA干扰载体 嘌呤霉素 puro
发布时间:2018-05-28 11:15:00 | 浏览次数:


测序及更多信息: 

https://pan.baidu.com/s/1GsonEdageMUdUEzaXiuaMQ 密码:xil1

采购地址:淘宝店铺地址

载体基本信息

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,shRNARNA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

稳定性:

/

组成型:

--

病毒/非病毒:

慢病毒



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

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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