pLVX-shRNA2慢病毒基因干扰载体shRNA表达质粒 BioVector NTCC质粒载体菌种细胞基因保藏中心

pLVX-shRNA2慢病毒基因干扰载体shRNA表达质粒

BioVector NTCC质粒载体菌种细胞基因保藏中心                      

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
          www.biovector.net
COMMENT     VNTAUTHORNAME|www.biovector.net|
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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