pMetLuc2-Control

价格:2600元

联系方式:I47-825O-882O

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载体基本信息

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

载体质粒图谱和多克隆位点信息

pMetLuc2-Control载体图谱



pMetLuc2-Control 载体特征

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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