pLVX-shRNA2

价格:1300元

联系方式:I47-825O-882O

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载体基本信息

出品公司: Clontech
载体名称: pLVX-shRNA2 , pLVX shRNA2
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: 7881 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: ZsGreen1
备注: 含有人U6启动子的慢病毒shRNA载体
产品目录号: 632179
稳定性: /
组成型: --
病毒/非病毒: 慢病毒

载体质粒图谱和多克隆位点信息

pLVX-shRNA2载体图谱


pLVX-shRNA2多克隆位点
pLVX-shRNA2载体特征

载体简介

Description
pLVX-shRNA2 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 driven by the RNA Pol III-dependent, human U6 promoter (PU6), located just upstream of the MCS.pLVX-shRNA2 can 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.

In addition to expressing shRNAs, pLVX-shRNA2 also expresses the fluorescent protein ZsGreen1, a human codon-optimized variant of the reef coral Zoanthus sp. green fluorescent protein, ZsGreen (1). Expression of ZsGreen1 is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), allowing it to be used as an indicator of transfection or transduction efficiency, as well as a marker for cell sorting.

pLVX-shRNA2 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 (2), 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 (3). Finally, pLVX-shRNA2 also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, this element creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (4). The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.

Use
Before pLVX-shRNA2 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 (5).

ZsGreen1 is the brightest commercially available green fluorescent protein. The presence of this protein allows transductants to be sorted by flow cytometry with standard FITC filter sets (ZsGreen1 has an excitation maximum of 493 nm and an emission maximum of 505 nm).

载体序列

LOCUS       pLVX-shRNA2   7880 bp    DNA   circular             8-NOV-2009
COMMENT     Created by Clontech Laboratories Inc.
COMMENT     Cat. No. 632179
COMMENT     This file is created by Vector NTI
            http://www.biofeng.com/
COMMENT     VNTAUTHORNAME|biofeng.com|
FEATURES             Location/Qualifiers
     misc_feature    2187..2444
                     /label=Human\U6\promoter
     misc_feature    2462..3050
                     /label=CMV\IE\Promoter
     misc_feature    3074..3769
                     /label=ZsGreen1
     LTR             1..635
                     /label=5'\LTR
     misc_feature    685..822
                     /label=packaging\signal
     misc_feature    2028..2151
                     /label=cPPT/CTS\(DNA\flap)
     misc_feature    3802..4393
                     /label=WPRE
     rep_origin      complement(5701..6374)
                     /label=pUC\origin
     CDS             complement(6519..7515)
                     /label=Amp(R)
     misc_binding    1303..1536
                     /label=RRE
     splicing_signal 744..745
                     /label=SD
     splicing_signal 1883..1884
                     /label=SA
     LTR             551..635
                     /label=U5
     misc_binding    636..653
                     /label=PBS
     LTR             4596..5232
                     /label=3'\LTR
BASE COUNT     2086 a      1881 c      1985 g      1928 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 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 
     2521 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 
     2581 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 
     2641 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 
     2701 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 
     2761 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 
     2821 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 
     2881 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 
     2941 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 
     3001 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct 
     3061 accggtcgcc accatggccc agtccaagca cggcctgacc aaggagatga ccatgaagta 
     3121 ccgcatggag ggctgcgtgg acggccacaa gttcgtgatc accggcgagg gcatcggcta 
     3181 ccccttcaag ggcaagcagg ccatcaacct gtgcgtggtg gagggcggcc ccttgccctt 
     3241 cgccgaggac atcttgtccg ccgccttcat gtacggcaac cgcgtgttca ccgagtaccc 
     3301 ccaggacatc gtcgactact tcaagaactc ctgccccgcc ggctacacct gggaccgctc 
     3361 cttcctgttc gaggacggcg ccgtgtgcat ctgcaacgcc gacatcaccg tgagcgtgga 
     3421 ggagaactgc atgtaccacg agtccaagtt ctacggcgtg aacttccccg ccgacggccc 
     3481 cgtgatgaag aagatgaccg acaactggga gccctcctgc gagaagatca tccccgtgcc 
     3541 caagcagggc atcttgaagg gcgacgtgag catgtacctg ctgctgaagg acggtggccg 
     3601 cttgcgctgc cagttcgaca ccgtgtacaa ggccaagtcc gtgccccgca agatgcccga 
     3661 ctggcacttc atccagcaca agctgacccg cgaggaccgc agcgacgcca agaaccagaa 
     3721 gtggcacctg accgagcacg ccatcgcctc cggctccgcc ttgccctaac tcgagtctag 
     3781 agggcccgac gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact 
     3841 ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg 
     3901 tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg 
     3961 ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg 
     4021 tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct cctttccggg 
     4081 actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc 
     4141 tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg 
     4201 acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc 
     4261 tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct 
     4321 ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc 
     4381 gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa 
     4441 gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg 
     4501 aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg 
     4561 tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac 
     4621 gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc 
     4681 agaactacac accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc 
     4741 tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt 
     4801 tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt 
     4861 ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga 
     4921 actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg 
     4981 gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt 
     5041 gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta 
     5101 gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc 
     5161 cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa 
     5221 atctctagca gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa 
     5281 tgaatatcag agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag 
     5341 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt 
     5401 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc 
     5461 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc 
     5521 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct 
     5581 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 
     5641 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 
     5701 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 
     5761 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 
     5821 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 
     5881 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 
     5941 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 
     6001 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 
     6061 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 
     6121 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 
     6181 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 
     6241 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 
     6301 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 
     6361 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 
     6421 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 
     6481 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 
     6541 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 
     6601 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 
     6661 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 
     6721 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 
     6781 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 
     6841 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 
     6901 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 
     6961 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 
     7021 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 
     7081 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 
     7141 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 
     7201 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 
     7261 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 
     7321 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 
     7381 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 
     7441 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 
     7501 gtgccacctg acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta 
     7561 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag 
     7621 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca 
     7681 agctaaccaa aatcatccca aacttcccac cccataccct attaccactg ccaattacct 
     7741 gtggtttcat ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg 
     7801 tatttgttaa atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata 
     7861 tgtactacaa acttagtagt 
//

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