| 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 |
买家导航
pLVX-shRNA2载体质粒基本信息
pLVX-shRNA2质粒图谱载体图谱和pLVX-shRNA2载体序列质粒序列多克隆位点信息
pLVX-shRNA2质粒载体简介
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).
pLVX-shRNA2质粒序列载体序列
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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