| 出品公司: | Clontech |
|---|---|
| 载体名称: | pMetLuc2-Control |
| 质粒类型: | 慢病毒载体；无启动子载体；荧光素酶报告载体 |
| 高拷贝/低拷贝: | 高拷贝 |
| 克隆方法: | 限制性内切酶，多克隆位点 |
| 启动子: | CMV |
| 载体大小: | 4784 bp |
| 5' 测序引物及序列: | CMV Forward： CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物及序列: | -- |
| 载体标签: | -- |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | 新霉素（Neomycin） |
| 克隆菌株: | DH5alpha等 |
| 宿主细胞（系）: | 常规细胞系（293、CV-1、CHO等） |
| 备注: |
pMetLuc2-Control组成型表达荧光素酶MetLuc，作为pMetLuc2-Reporter载体的阳性对照； MetLuc是已知最小分子量的荧光素酶； 荧光素酶报告基因可以用来定性定量分析启动子及其它顺式转录元件的活性。 |
| 产品目录号: | 631729 |
| 稳定性: | 瞬表达 或 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
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
pMetLuc2-Control载体描述
The pMetLuc2-Control vector encodes a sequence-optimized, secreted luciferase from the marine copepod Metridia longa. The 24 kDa Metridia luciferase (MetLuc) contains a 17 amino acid, N-terminal signal peptide that allows the protein to be efficiently secreted into the cell culture medium (1), making it easy to detect without cell lysis.Expression of MetLuc is driven by the constitutively active cytomegalovirus immediate early promoter (PCMV IE). SV40 polyadenylation signals downstream of the MetLuc gene direct proper processing of the 3' end of the MetLuc mRNA. The vector backbone contains an SV40 origin for replication in mammalian cells expressing the SV40 large T antigen, a pUC origin of replication for propagation in E. coli, and an f1 origin for single-stranded DNA production. A neomycin-resistance cassette (Neor) allows stably transfected eukaryotic cells to be selected using G418. This cassette consists of the SV40 early promoter, the Tn5 kanamycin/neomycin resistance gene, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV TK) gene. The vector also contains a synthetic transcription blocker (TB), composed of adjacent polyadenylation and transcription pause sites, that reduces background readthrough transcription (2). A bacterial promoter (PKanr) upstream of the cassette allows kanamycin resistance in E. coli.
The pMetLuc2-Control vector can be used as a positive control for the constitutive expression and secretion of MetLuc in experiments using the pMetLuc2-Reporter vector. pMetLuc2-Control can also be used to monitor the inhibitory effect of drugs, such as Brefeldin A, on protein secretion.
The pMetLuc2-Control Vector can be transfected into mammalian cells using any standard transfection method. Stable transfectants can be selected using G418 when required (3). The presence of MetLuc in the cell culture medium can be easily detected by adding luciferase substrate to a small aliquot of the medium and analyzing the sample with a luminometer.
Propagation in E. coli
Suitable host strains: DH5α, HB101, and other general purpose strains. Single-stranded DNA production
requires a host containing an F plasmid such as JM109 or XL1-Blue.
Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
E. coli replication origin: pUC
Plasmid incompatibility group: pMB1/Col E1
Note: The attached sequence file has been 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.
References
1. Markova, S.V. et al. (2004) J. Bio. Chem. 279(5):3212-3217.
2. Eggermont, J. & Proudfoot, N. (1993) EMBO J. 12(6):2539–2548.
3. Gorman, C. (1985) DNA cloning: A Practical Approach, Vol. II. Ed. D. M. Glover. (IRL Press Oxford, U.K.), pp. 143-190.
载体序列
LOCUS pMetLuc2-Control 4781 bp DNA circular 8-NOV-2009
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat. No. 631735,631731,631743,631744,631732,631742,631729,631745,631730
FEATURES Location/Qualifiers
CDS 2517..3311
/label=Kan(R)/Neo(R)
promoter 1..589
/label=CMV\IE\Promoter
CDS 628..1285
/label=Metridia\luciferase
misc_feature 4569..4722
/label=Transcription\blocker
polyA_signal 1440..1445
/label=SV40\early\poly\A\signal
polyA_signal 1469..1474
/label=SV40\early\polyA\signal
polyA_signal 3547..3552
/label=HSV\TK\polyA\signal
polyA_signal 3560..3565
/label=HSV\TK\polyA\signal
rep_origin complement(1537..1992)
/label=f1\origin
rep_origin 3896..4539
/label=pUC\origin
rep_origin 2333..2471
/label=SV40\origin
misc_feature 2388..2394
/label=SV40\early\Promoter
promoter 2054..2082
/label=Kan(R)\Promoter
BASE COUNT 1171 a 1268 c 1253 g 1089 t
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atccaccggt cgccaccatg gacatcaagg tggtgttcac cctggtgttc
661 agcgccctgg tgcaggccaa gagcaccgag ttcgacccca acatcgacat cgtgggcctg
721 gaaggcaagt tcggcatcac caacctggaa accgacctgt tcaccatctg ggagaccatg
781 gaagtgatga tcaaggccga catcgccgac accgaccggg ccagcaactt cgtggccacc
841 gagaccgacg ccaaccgggg caagatgccc ggcaagaagc tgcccctggc cgtcatcatg
901 gaaatggaag ccaacgcctt caaggccggc tgcacccggg gctgcctgat ctgcctgagc
961 aagatcaagt gcaccgccaa gatgaaggtg tacatccccg gcaggtgcca cgactacggc
1021 ggcgacaaga aaaccggcca ggccggcatc gtgggcgcca tcgtggacat ccccgagatc
1081 agcggcttca aagaaatggc ccccatggaa cagttcatcg cccaggtgga cagatgcgcc
1141 agctgcacca ccggctgcct gaagggcctg gccaacgtga agtgcagcga gctgctgaag
1201 aagtggctgc ccgaccgctg cgccagcttc gccgacaaga tccagaaaga ggtgcacaac
1261 atcaagggca tggccggcga caggtgagcg gccgcgactc tagatcataa tcagccatac
1321 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
1381 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
1441 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
1501 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
1561 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
1621 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
1681 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
1741 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
1801 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
1861 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
1921 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
1981 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
2041 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
2101 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
2161 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
2221 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
2281 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
2341 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
2401 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
2461 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
2521 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
2581 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
2641 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
2701 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
2761 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
2821 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
2881 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
2941 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
3001 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
3061 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
3121 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
3181 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
3241 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
3301 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
3361 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
3421 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
3481 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
3541 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
3601 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
3661 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
3721 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
3781 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
3841 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
3901 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
3961 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
4021 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
4081 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
4141 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
4201 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
4261 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
4321 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
4381 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
4441 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
4501 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
4561 tcacatgtaa taaaatatct ttattttcat tacatctgtg tgttggtttt ttgtgtgaat
4621 cgatagtact aacatacgct ctccatcaaa acaaaacgaa acaaaacaaa ctagcaaaat
4681 aggctgtccc cagtgcaagt gcaggtgcca gaacatttct ctacatgttc tttcctgcgt
4741 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
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