| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-IRES-Neo, pLVX IRES Neo |
| 质粒类型: | 哺乳动物细胞慢病毒表达载体 |
| 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: | 8269 bp |
| 5' 测序引物及序列: | CMV-F: CGCAAATGGGCGGTAGGCGTG (Invitrogen) |
| 3' 测序引物及序列: | IRES-R: CCTCACATTGCCAAAAGACG |
| 载体标签: | -- |
| 载体抗性: | 氨苄 |
| 筛选标记: | 新霉素(G418) |
| 备注: | 载体含有新霉素筛选标记。 |
| 产品目录号: | 632181 |
| 稳定性: | / |
| 组成型: | 诱导型 |
| 病毒/非病毒: | 慢病毒 |
pLVX-IRES-Neo慢病毒表达载体质粒图谱序列抗性价格报价Biovector NTCC Inc.
- 价 格：¥1920
- 货 号：BiovectorLVX-IRES-Neo
- 产 地：北京
- BioVector NTCC典型培养物保藏中心
- 联系人：Dr.Xu, Biovector NTCC Inc.
电话：400-800-2947 工作QQ:1843439339 (微信同号)
邮件：Biovector@163.com
手机：18901268599
地址：北京
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载体基本信息
载体质粒图谱和多克隆位点信息
载体简介
Description
pLVX-IRES-Neo is an HIV-1-based, lentiviral expression vector that allows the simultaneous expression of your protein of interest and G418 resistance (Neor) in virtually any mammalian cell type, including primary cells. The vector expresses your protein of interest and Neor from a bicistronic mRNA transcript, allowing G418 resistance to be used as an indicator of transduction efficiency and a marker for selection.
Expression of the bicistronic transcript is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE) located just upstream of the MCS. An encephalomyocarditis virus (EMCV) internal ribosome entry site (IRES), positioned between the MCS and Neor, facilitates cap-independent translation of Neor from an internal start site at the IRES/Neor junction (1).
pLVX-IRES-Neo contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer, transgene expression, 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-IRES-Neo 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
pLVX-IRES-Neo is available as part of the Lenti-X™ Bicistronic Expression System (Neo; Cat. No. 632181). The vector is designed to constitutively coexpress your protein of interest and G418 resistance from PCMV IE
when transduced into mammalian cells. Before it can be transduced into target cells, the vector must be packaged into viral particles in HEK293T cells, using our Lenti-X™ HTX Packaging System (Cat. Nos. 631247 and 631249). 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 nondividing and primary cells (5).
Selection of Stable Transductants
• Selectable marker: vector confers resistance to G418.
Propagation in E. coli
• Suitable host strains: DH5α™, DH10B™ and other general purpose strains.
• Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
• E. coli replication origin: pUC
• Copy number: high
Notes:
The vector sequence was compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequenced.
The viral supernatants produced by this lentiviral vector could contain potentially hazardous recombinant virus. Due caution must be exercised in the production and handling of recombinant lentivirus. Appropriate NIH, regional, and institutional guidelines apply.
References
1. Jang, S. K. et al. (1988) J. Virol. 62(8):2636–2643.
2. Zufferey, R. et al. (1999) J. Virol. 73(4):2886–2892.
3. Cochrane, A. W. et al. (1990) Proc. Natl. Acad. Sci. USA 87(3):1198–1202.
4. Zennou, V. et al. (2000) Cell 101(2):173–185.
5. Wu, X. et al. (2000) Mol. Ther. 2(1):47–55.
载体序列
LOCUS pLVX-IRES-Neo 8269 bp DNA circular 8-NOV-2009
COMMENT Created by Clontech Laboratories
COMMENT This file is created by Vector NTI
http://www.biovector.net/
COMMENT VNTAUTHORNAME|biovector.net|
FEATURES Location/Qualifiers
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=packaging\signal
misc_feature 2028..2151
/label=cPPT/CTS\(DNA\flap)
promoter 2185..2787
/label=CMV\IE\Promoter
misc_feature 4238..4829
/label=WPRE
rep_origin complement(6137..6810)
/label=pUC\origin
CDS complement(6955..7951)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
CDS 3430..4224
/label=Neo(R)
misc_structure 2841..3429
/label=IRES
misc_feature 2803..2840
/label=MCS
LTR 5032..5668
/label=3'\LTR
BASE COUNT 2115 a 2002 c 2130 g 2022 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 cgataagctt gggagttccg cgttacataa cttacggtaa
2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
2761 tccatagaag acaccgactc tactagagga tctatttccg gtgaattcct cgagactagt
2821 tctagagcgg ccgcggatcc cgcccctctc cctccccccc ccctaacgtt actggccgaa
2881 gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt
2941 cttttggcaa tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg
3001 gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc
3061 ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc
3121 ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa
3181 aggcggcaca accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc
3241 tctcctcaag cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg
3301 gatctgatct ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac
3361 gtctaggccc cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt
3421 gccacaacca tggctgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
3481 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
3541 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
3601 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
3661 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
3721 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
3781 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
3841 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
3901 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
3961 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
4021 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
4081 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
4141 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
4201 cgccttcttg acgagttctt ctgaacgcgt ctggaacaat caacctctgg attacaaaat
4261 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
4321 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
4381 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
4441 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
4501 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
4561 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
4621 gttgtcgggg aagctgacgt cctttccatg gctgctcgcc tgtgttgcca cctggattct
4681 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
4741 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
4801 gatctccctt tgggccgcct ccccgcctgg aattaattct gcagtcgaga cctagaaaaa
4861 catggagcaa tcacaagtag caatacagca gctaccaatg ctgattgtgc ctggctagaa
4921 gcacaagagg aggaggaggt gggttttcca gtcacacctc aggtaccttt aagaccaatg
4981 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagagggg actggaaggg
5041 ctaattcact cccaacgaag acaagatatc cttgatctgt ggatctacca cacacaaggc
5101 tacttccctg attagcagaa ctacacacca gggccagggg tcagatatcc actgaccttt
5161 ggatggtgct acaagctagt accagttgag ccagataagg tagaagaggc caataaagga
5221 gagaacacca gcttgttaca ccctgtgagc ctgcatggga tggatgaccc ggagagagaa
5281 gtgttagagt ggaggtttga cagccgccta gcatttcatc acgtggcccg agagctgcat
5341 ccggagtact tcaagaactg ctgatatcga gcttgctaca agggactttc cgctggggac
5401 tttccaggga ggcgtggcct gggcgggact ggggagtggc gagccctcag atcctgcata
5461 taagcagctg ctttttgcct gtactgggtc tctctggtta gaccagatct gagcctggga
5521 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
5581 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
5641 ttagtcagtg tggaaaatct ctagcagtag tagttcatgt catcttatta ttcagtattt
5701 ataacttgca aagaaatgaa tatcagagag tgagaggcct tgacattgct agcgtttacc
5761 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
5821 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
5881 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
5941 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
6001 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
6061 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
6121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
6181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
6241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
6301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
6361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
6421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
6481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
6541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
6601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
6661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
6721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
6781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
6841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
6901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
6961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
7021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
7081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
7141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
7201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
7261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
7321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
7381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
7441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
7501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
7561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
7621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
7681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
7741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
7801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
7861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
7921 cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatcaac ttgtttattg
7981 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
8041 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
8101 tcaactggat aactcaagct aaccaaaatc atcccaaact tcccacccca taccctatta
8161 ccactgccaa ttacctgtgg tttcatttac tctaaacctg tgattcctct gaattatttt
8221 cattttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
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