| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-DsRed-Express2-N1 |
| 质粒类型: | 慢病毒表达载体；荧光报告载体 |
| 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: | 8745 bp |
| 5' 测序引物及序列: | CMV-F: CGCAAATGGGCGGTAGGCGTG (Invitrogen) |
| 3' 测序引物及序列: | -- |
| 载体标签: | DsRed-Express2（C-端） |
| 载体抗性: | 氨苄青霉素 |
| 筛选标记: | 嘌呤霉素(Puromycin) |
| 克隆菌株: | Stbl3 |
| 宿主细胞（系）: | 常规细胞系，293、CV-1、CHO等 |
| 备注: |
pLVX-DsRed-Express2-N1载体表达C端DsRed-Express2融合蛋白； DsRed-Express2是DsRed的突变体， 与DsRed相比，DsRed-Express2细胞毒性低，尤其适用于原代细胞和干细胞的荧光标记 |
| 产品目录号: | 632560 |
| 稳定性: | 稳表达 |
| 组成型: | 组成型 |
| 病毒/非病毒: | 慢病毒 |
买家导航
pLVX-DsRed-Express2-N1载体质粒基本信息
pLVX-DsRed-Express2-N1质粒图谱载体图谱和pLVX-DsRed-Express2-N1载体序列质粒序列多克隆位点信息
pLVX-DsRed-Express2-N1质粒载体简介
Description
pLVX-DsRed-Express2-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to DsRed-Express2, a mutant fluorescent protein derived from the tetrameric Discosoma sp. red fluorescent protein, DsRed (1). Genes cloned into the multiple cloning site (MCS), located upstream of the DsRed-Express2 coding sequence, are expressed as N-terminal DsRed-Express2 fusion proteins. Expression of the fusion protein
is driven by the constitutively active human cytomegalovirus immediate early promoter (PCMV IE), located just upstream of the MCS. Lentiviral particles derived from the vector allow the expression of DsRed-Express2 fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses DsRed-Express2, and may be used to produce marker virus to optimize infection protocols.
pLVX-DsRed-Express2-N1 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 and transgene RNA (2), leading to increased viral titers from packaging cells, and enhanced expression of your gene of interest in target 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-DsRed-Express2-N1 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).
In addition to lentiviral elements, pLVX-DsRed-Express2-N1 contains a puromycin resistance gene (Puror) under the control of the murine phosphoglycerate kinase (PGK) 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
To construct a fusion protein, the gene of interest must be cloned into pLVX-DsRed-Express2-N1 so that it is in-frame with the DsRed-Express2 coding sequence. The inserted sequence should include an initiation codon (ATG) and lack in-frame stop codons.
The fusion protein is constitutively expressed when pLVX-DsRed-Express2-N1 is transduced into target cells. Before the vector can be transduced, however, it must be transfected into 293T packaging cells with our Lenti-X™ HT Packaging System (Cat. Nos. 632160 and 632161). This packaging system allows you to safely produce 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).
pLVX-DsRed-Express2-N1质粒序列载体序列
LOCUS pLVX-DsRed-Express2-N1 8745 bp DNA circular 21-OCT-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 632560
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT VNTAUTHORNAME|biofeng.com|
FEATURES Location/Qualifiers
LTR 1..635
/label=5'\LTR
misc_feature 685..822
/label=packaging\signal
misc_feature 4713..5304
/label=WPRE
CDS complement(7431..8427)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
LTR 5507..6143
/label=3'\LTR
promoter 3571..4079
/label=PGK\Promoter
CDS 4100..4699
/label=Puro(R)
CDS 2875..3552
/label=DsRed-Express2
misc_feature 2028..2151
/label=cPPT/CTS\(DNA\flap)
promoter 2185..2787
/label=CMV\IE\Promoter
rep_origin complement(6613..7286)
/label=pUC\Ori
misc_feature 2815..2867
/label=MCS
BASE COUNT 2160 a 2270 c 2317 g 1998 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 tcgctagcgc taccggactc agatctcgag
2821 ctcaagcttc gaattctgca gtcgacggta ccgcgggccc gggatccacc ggtcatggat
2881 agcactgaga acgtcatcaa gcccttcatg cgcttcaagg tgcacatgga gggctccgtg
2941 aacggccacg agttcgagat cgagggcgag ggcgagggca agccctacga gggcacccag
3001 accgccaagc tgcaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc
3061 ccccagttcc agtacggctc caaggtgtac gtgaagcacc ccgccgacat ccccgactac
3121 aagaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
3181 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcaccttcat ctaccacgtg
3241 aagttcatcg gcgtgaactt cccctccgac ggccccgtaa tgcagaagaa gactctgggc
3301 tgggagccct ccaccgagcg cctgtacccc cgcgacggcg tgctgaaggg cgagatccac
3361 aaggcgctga agctgaaggg cggcggccac tacctggtgg agttcaagtc aatctacatg
3421 gccaagaagc ccgtgaagct gcccggctac tactacgtgg actccaagct ggacatcacc
3481 tcccacaacg aggactacac cgtggtggag cagtacgagc gcgccgaggc ccgccaccac
3541 ctgttccagt gagcggccgc gactctagat aattctaccg ggtaggggag gcgcttttcc
3601 caaggcagtc tggagcatgc gctttagcag ccccgctggg cacttggcgc tacacaagtg
3661 gcctctggcc tcgcacacat tccacatcca ccggtaggcg ccaaccggct ccgttctttg
3721 gtggcccctt cgcgccacct tctactcctc ccctagtcag gaagttcccc cccgccccgc
3781 agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg tctcactagt ctcgtgcaga
3841 tggacagcac cgctgagcaa tggaagcggg taggcctttg gggcagcggc caatagcagc
3901 tttgctcctt cgctttctgg gctcagaggc tgggaagggg tgggtccggg ggcgggctca
3961 ggggcgggct caggggcggg gcgggcgccc gaaggtcctc cggaggcccg gcattctgca
4021 cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc ctcatctccg ggcctttcga
4081 cctgcagccc aagcttacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga
4141 cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg
4201 ccacaccgtc gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct
4261 cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc
4321 ggtctggacc acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg
4381 catggccgag ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc
4441 gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca
4501 ccagggcaag ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc
4561 cggggtgccc gccttcctgg agacctccgc gccccgcaac ctccccttct acgagcggct
4621 cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac
4681 ccgcaagccc ggtgcctgac cgcgtctgga acaatcaacc tctggattac aaaatttgtg
4741 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
4801 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
4861 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
4921 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
4981 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
5041 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
5101 cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg attctgcgcg
5161 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
5221 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
5281 ccctttgggc cgcctccccg cctggaatta attctgcagt cgagacctag aaaaacatgg
5341 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
5401 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
5461 caaggcagct gtagatctta gccacttttt aaaagaaaag aggggactgg aagggctaat
5521 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
5581 ccctgattag cagaactaca caccagggcc aggggtcaga tatccactga cctttggatg
5641 gtgctacaag ctagtaccag ttgagccaga taaggtagaa gaggccaata aaggagagaa
5701 caccagcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtgtt
5761 agagtggagg tttgacagcc gcctagcatt tcatcacgtg gcccgagagc tgcatccgga
5821 gtacttcaag aactgctgat atcgagcttg ctacaaggga ctttccgctg gggactttcc
5881 agggaggcgt ggcctgggcg ggactgggga gtggcgagcc ctcagatcct gcatataagc
5941 agctgctttt tgcctgtact gggtctctct ggttagacca gatctgagcc tgggagctct
6001 ctggctaact agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
6061 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
6121 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
6181 ttgcaaagaa atgaatatca gagagtgaga ggccttgaca ttgctagcgt tttaccgtcg
6241 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
6301 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
6361 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
6421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
6481 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
6541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
6601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
6661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
6721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
6781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
6841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
6901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
6961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
7021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
7081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
7141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
7201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
7261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
7321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
7381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
7561 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
8281 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
8341 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
8401 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcaacttgt ttattgcagc
8461 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
8521 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctggatcaa
8581 ctggataact caagctaacc aaaatcatcc caaacttccc accccatacc ctattaccac
8641 tgccaattac ctgtggtttc atttactcta aacctgtgat tcctctgaat tattttcatt
8701 ttaaagaaat tgtatttgtt aaatatgtac tacaaactta gtagt
//
