| 出品公司: | Clontech |
|---|---|
| 载体名称: | pLVX-AmCyan1-N1, pLVX AmCyan1 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 IE |
| 克隆方法: | 多克隆位点，限制性内切酶 |
| 载体大小: | 8757 bp |
| 5' 测序引物及序列: | -- |
| 3' 测序引物及序列: | -- |
| 载体标签: | AmCyan1（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-AmCyan1-N1载体表达C端AmCyan1融合蛋白； 青色荧光蛋白AmCyan1具有高度可溶性，高亮度，同时与其它荧光蛋白颜色区别大。 |
| 产品目录号: | 632558 |
| 稳定性: | 稳表达 |
| 组成型: | 组成型 |
| 病毒/非病毒: | 慢病毒 |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
pLVX-AmCyan1-N1载体描述
pLVX-AmCyan1-N1 is an HIV-1-based, lentiviral expression vector that allows you to express your gene of interest fused to AmCyan1, a human codon-optimized variant of the wild-type Anemonia majano cyan fluorescent protein (1). Genes cloned into the multiple cloning site (MCS), located upstream of the AmCyan1 coding sequence, are expressed as N-terminal AmCyan1 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 AmCyan1 fusion proteins in virtually any cell type, including primary cells. The unmodified vector expresses AmCyan1, and may be used to produce marker virus to optimize infection protocols.
pLVX-AmCyan1-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-AmCyan1-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-AmCyan1-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.
To construct a fusion protein, the gene of interest must be cloned into pLVX-AmCyan1-N1 so that it is in-frame with the AmCyan1 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-AmCyan1-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).
Selection of Stable Transfectants
Selectable marker: plasmid confers resistance to puromycin.
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
Excitation and emission maxima of AmCyan1
Excitation maximum = 458 nm
Emission maximum = 489 nm
载体序列
LOCUS pLVX-AmCyan1-N1 8757 bp DNA circular 7-OCT-2009
COMMENT Created by Clontech Laboratories Inc.
COMMENT Cat. No. 632558
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 4725..5316
/label=WPRE
CDS complement(7443..8439)
/label=Amp(R)
misc_binding 1303..1536
/label=RRE
misc_binding 636..653
/label=PBS
LTR 5519..6155
/label=3'\LTR
promoter 3583..4091
/label=PGK\Promoter
CDS 4112..4711
/label=Puro(R)
CDS 2875..3564
/label=AmCyan1
misc_feature 2028..2151
/label=cPPT/CTS\(DNA\flap)
promoter 2185..2787
/label=CMV\IE\Promoter
rep_origin complement(6625..7298)
/label=pUC\ori
misc_feature 2815..2867
/label=MCS
BASE COUNT 2163 a 2296 c 2300 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 ggtcatggcc
2881 ctgtccaaca agttcatcgg cgacgacatg aagatgacct accacatgga cggctgcgtg
2941 aacggccact acttcaccgt gaagggcgag ggcagcggca agccctacga gggcacccag
3001 acctccacct tcaaggtgac catggccaac ggcggccccc tggccttctc cttcgacatc
3061 ctgtccaccg tgttcatgta cggcaaccgc tgcttcaccg cctaccccac cagcatgccc
3121 gactacttca agcaggcctt ccccgacggc atgtcctacg agagaacctt cacctacgag
3181 gacggcggcg tggccaccgc cagctgggag atcagcctga agggcaactg cttcgagcac
3241 aagtccacct tccacggcgt gaacttcccc gccgacggcc ccgtgatggc caagaagacc
3301 accggctggg acccctcctt cgagaagatg accgtgtgcg acggcatctt gaagggcgac
3361 gtgaccgcct tcctgatgct gcagggcggc ggcaactaca gatgccagtt ccacacctcc
3421 tacaagacca agaagcccgt gaccatgccc cccaaccacg tggtggagca ccgcatcgcc
3481 agaaccgacc tggacaaggg cggcaacagc gtgcagctga ccgagcacgc cgtggcccac
3541 atcacctccg tggtgccctt ctgagcggcc gcgactctag ataattctac cgggtagggg
3601 aggcgctttt cccaaggcag tctggagcat gcgctttagc agccccgctg ggcacttggc
3661 gctacacaag tggcctctgg cctcgcacac attccacatc caccggtagg cgccaaccgg
3721 ctccgttctt tggtggcccc ttcgcgccac cttctactcc tcccctagtc aggaagttcc
3781 cccccgcccc gcagctcgcg tcgtgcagga cgtgacaaat ggaagtagca cgtctcacta
3841 gtctcgtgca gatggacagc accgctgagc aatggaagcg ggtaggcctt tggggcagcg
3901 gccaatagca gctttgctcc ttcgctttct gggctcagag gctgggaagg ggtgggtccg
3961 ggggcgggct caggggcggg ctcaggggcg gggcgggcgc ccgaaggtcc tccggaggcc
4021 cggcattctg cacgcttcaa aagcgcacgt ctgccgcgct gttctcctct tcctcatctc
4081 cgggcctttc gacctgcagc ccaagcttac catgaccgag tacaagccca cggtgcgcct
4141 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
4201 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
4261 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
4321 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
4381 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
4441 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt
4501 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
4561 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
4621 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
4681 ctggtgcatg acccgcaagc ccggtgcctg accgcgtctg gaacaatcaa cctctggatt
4741 acaaaatttg tgaaagattg actggtattc ttaactatgt tgctcctttt acgctatgtg
4801 gatacgctgc tttaatgcct ttgtatcatg ctattgcttc ccgtatggct ttcattttct
4861 cctccttgta taaatcctgg ttgctgtctc tttatgagga gttgtggccc gttgtcaggc
4921 aacgtggcgt ggtgtgcact gtgtttgctg acgcaacccc cactggttgg ggcattgcca
4981 ccacctgtca gctcctttcc gggactttcg ctttccccct ccctattgcc acggcggaac
5041 tcatcgccgc ctgccttgcc cgctgctgga caggggctcg gctgttgggc actgacaatt
5101 ccgtggtgtt gtcggggaag ctgacgtcct ttccatggct gctcgcctgt gttgccacct
5161 ggattctgcg cgggacgtcc ttctgctacg tcccttcggc cctcaatcca gcggaccttc
5221 cttcccgcgg cctgctgccg gctctgcggc ctcttccgcg tcttcgcctt cgccctcaga
5281 cgagtcggat ctccctttgg gccgcctccc cgcctggaat taattctgca gtcgagacct
5341 agaaaaacat ggagcaatca caagtagcaa tacagcagct accaatgctg attgtgcctg
5401 gctagaagca caagaggagg aggaggtggg ttttccagtc acacctcagg tacctttaag
5461 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa agaggggact
5521 ggaagggcta attcactccc aacgaagaca agatatcctt gatctgtgga tctaccacac
5581 acaaggctac ttccctgatt agcagaacta cacaccaggg ccaggggtca gatatccact
5641 gacctttgga tggtgctaca agctagtacc agttgagcca gataaggtag aagaggccaa
5701 taaaggagag aacaccagct tgttacaccc tgtgagcctg catgggatgg atgacccgga
5761 gagagaagtg ttagagtgga ggtttgacag ccgcctagca tttcatcacg tggcccgaga
5821 gctgcatccg gagtacttca agaactgctg atatcgagct tgctacaagg gactttccgc
5881 tggggacttt ccagggaggc gtggcctggg cgggactggg gagtggcgag ccctcagatc
5941 ctgcatataa gcagctgctt tttgcctgta ctgggtctct ctggttagac cagatctgag
6001 cctgggagct ctctggctaa ctagggaacc cactgcttaa gcctcaataa agcttgcctt
6061 gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc tggtaactag agatccctca
6121 gaccctttta gtcagtgtgg aaaatctcta gcagtagtag ttcatgtcat cttattattc
6181 agtatttata acttgcaaag aaatgaatat cagagagtga gaggccttga cattgctagc
6241 gttttaccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
6301 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
6361 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
6421 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
6481 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
6541 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
6601 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
6661 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
6721 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
6781 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
6841 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
6901 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
6961 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
7021 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
7081 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
7141 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
7201 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
7261 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
7321 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
7381 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
7441 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
7501 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
7561 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
7621 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
7681 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
7741 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
7801 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
7861 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
7921 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
7981 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
8041 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
8101 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
8161 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
8221 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
8281 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
8341 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
8401 gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg agatcaactt
8461 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
8521 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
8581 tgtctggatc aactggataa ctcaagctaa ccaaaatcat cccaaacttc ccaccccata
8641 ccctattacc actgccaatt acctgtggtt tcatttactc taaacctgtg attcctctga
8701 attattttca ttttaaagaa attgtatttg ttaaatatgt actacaaact tagtagt
//
