出品公司: | Clontech |
---|---|
载体名称: | pLVX-DsRed-Express2-C1 |
质粒类型: | 慢病毒载体;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(N-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-C1载体表达N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: | 632559 |
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-C1载体质粒基本信息
pLVX-DsRed-Express2-C1质粒图谱载体图谱和pLVX-DsRed-Express2-C1载体序列质粒序列多克隆位点信息
pLVX-DsRed-Express2-C1质粒载体简介
Description
pLVX-DsRed-Express2-C1 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 at the C-terminal end of the DsRed-Express2 coding sequence, are expressed as C-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 DsRed-Express2 coding sequence. 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-C1 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-C1 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-C1 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-C1 so that it is in-frame with the DsRed-Express2 coding sequence. The inserted sequence does not require an initiation codon (ATG) or a stop codon (TAA, TAG, TGA); however, if you don't want to use the stop codons downstream of the MCS (see map), you can add a stop codon to the end of your gene of interest.
The fusion protein is constitutively expressed when pLVX-DsRed-Express2-C1 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-C1质粒序列载体序列
LOCUS pLVX-DsRed-Express2-C1 8745 bp DNA circular 7-OCT-2009 COMMENT Created by Clontech Laboratories Inc. COMMENT Cat. No. 632559 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) misc_feature 3556..3558 /label=Stop misc_feature 3560..3562 /label=Stop CDS 2807..3481 /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 3564..3566 /label=Stop misc_feature 3495..3547 /label=MCS BASE COUNT 2163 a 2270 c 2312 g 2000 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 tctaccggtc gccaccatgg atagcactga 2821 gaacgtcatc aagcccttca tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca 2881 cgagttcgag atcgagggcg agggcgaggg caagccctac gagggcaccc agaccgccaa 2941 gctgcaggtg accaagggcg gccccctgcc cttcgcctgg gacatcctgt ccccccagtt 3001 ccagtacggc tccaaggtgt acgtgaagca ccccgccgac atccccgact acaagaagct 3061 gtccttcccc gagggcttca agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt 3121 gaccgtgacc caggactcct ccctgcagga cggcaccttc atctaccacg tgaagttcat 3181 cggcgtgaac ttcccctccg acggccccgt aatgcagaag aagactctgg gctgggagcc 3241 ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag ggcgagatcc acaaggcgct 3301 gaagctgaag ggcggcggcc actacctggt ggagttcaag tcaatctaca tggccaagaa 3361 gcccgtgaag ctgcccggct actactacgt ggactccaag ctggacatca cctcccacaa 3421 cgaggactac accgtggtgg agcagtacga gcgcgccgag gcccgccacc acctgttcca 3481 gtccggactc agatctcgag ctcaagcttc gaattctgca gtcgacggta ccgcgggccc 3541 gggatccacc ggatctagat aactgatcat 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 //