pCRE-MetLuc2-Reporter

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: pCRE-MetLuc2-Reporter
质粒类型: 哺乳动物细胞表达载体;信号通路报告载体;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: TAL (TATA-like promoter)
载体大小: 4489 bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: --
载体抗性: Kanamycin.html' target='_blank'>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: 哺乳动物细胞表达载体pCRE-MetLuc2-Reporter是荧光素酶报告载体;
用于研究cAMP介导的信号通路;
MetLuc是已知分子量最小的荧光素酶;
荧光素酶报告基因既可以定性又可以定量。
产品目录号: 631745
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

pCRE-MetLuc2-Reporter载体图谱



pCRE-MetLuc2-Reporter 载体特征

载体简介

pCRE-MetLuc2-Reporter载体描述

pCRE-MetLuc2-Reporter allows you to monitor cAMP-mediated signal transduction directly from the cell culture medium, without cell lysis. The vector contains two copies of the cAMP response element (CRE; 1) fused to a TATA-like promoter (PTAL) region from the herpes simplex virus thymidine kinase (HSV-TK) gene. Located downstream of PTAL is a human codon optimized secreted-luciferase gene from the marine copepod Metridia longa. This gene encodes a 219 amino acid (24 kDa) polypeptide that includes a 17 amino acid N-terminal signal peptide necessary for secretion. Binding of transcription factors, such as the cAMP response element binding protein (CREB), to the CRE sequence allows Metridia luciferase (MetLuc) to be expressed and secreted into the surrounding medium (2).
To prevent read-through transcription of the Metridia luciferase gene, a synthetic transcription blocker (TB; 3, 4), composed of adjacent polyadenylation and transcription pause sites, is located upstream of the CRE. To ensure efficient processing of the luciferase transcript in eukaryotic cells, an SV40 early polyadenylation signal is located downstream of the Metridia luciferase coding sequence. The vector backbone also contains an f1 origin for single-stranded DNA production, a pUC origin of replication, and a kanamycin resistance gene for propagation and selection in E. coli.

pCRE-MetLuc2-Reporter is available as part of the Ready-To-Glow CRE Secreted Luciferase Reporter System (Cat. No. 631745). In mammalian cells containing pCRE-MetLuc2-Reporter, the addition of cAMP, or forskolin to the culture medium induces transcription factors, such as CREB or activating transcription factor (ATF) proteins, to bind to the CRE, thereby initiating transcription of the secreted luciferase reporter gene. Activation of cAMP-mediated signal transduction can be monitored simply by sampling the culture medium. To monitor the effect of a specific protein on the system, pCRE-MetLuc2-Reporter can be cotransfected with an expression
vector containing the gene of interest. pCRE-MetLuc2-Reporter can be transfected into mammalian cells using any standard transfection method. Stable transfectants can be selected using G418 when required.

Propagation in E. coli:
 Suitable host strains: DH5α, HB101 and other general purpose strains. Single-stranded DNA production
requires a host containing an F' episome, such as JM109 or XL1-Blue.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) to E. coli hosts.
 E. coli replication origin: pUC
 Plasmid incompatibility group: pMB1/Col E1

References:
1. Himmler, A., et al. (1993) J. Recep. Res. 13(1–4):79–94.
2. Markova, S. V., et al. (2004) J. Biol. Chem. 279(5):3212–3217.
3. Eggermont, J. & Proudfoot, N. (1993) EMBO J. 12(6):2539–2548.
4. Enriquez-Harris, P., et al. (1991) EMBO J. 10(7):1833–1842.

Note:
The vector sequence file has been compiled from information in the sequence databases, published literature, and other sources,
together with partial sequences obtained by Clontech Laboratories, Inc. This vector has not been completely sequenced.

载体序列

LOCUS       pCRE-MetLuc2-Reporter   4489 bp    DNA   circular             8-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631744,631745
FEATURES             Location/Qualifiers
     misc_feature    37..123
                     /label=cAMP\response\element\(CRE)
     promoter        130..278
                     /label=TATA\like\promoter\Ptal
     CDS             336..992
                     /label=Metridia\secreted\luciferase
     CDS             2225..3019
                     /label=Kan(R)/Neo(R)
     misc_feature    4277..4430
                     /label=Transcription\blocker
     polyA_signal    1148..1153
                     /label=SV40\early\polyA\signal
     rep_origin      complement(1245..1700)
                     /label=f1\origin
     rep_origin      2041..2179
                     /label=SV40\origin
     rep_origin      3604..4247
                     /label=pUC\origin
BASE COUNT     1097 a      1207 c      1193 g       992 t 
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgaggcac cagacagtga cgtcagctgc 
       61 cagatcccat ggccgtcata ctgtgacgtc tttcagacac cccattgacg tcaatgggag 
      121 aacagatctg ccgccccgac tgcatctgcg tgttcgaatt cgccaatgac aagacgctgg 
      181 gcggggtttg tgtcatcata gaactaaaga catgcaaata tatttcttcc ggggacaccg 
      241 ccagcaaacg cgagcaacgg gccacgggga tgaagcagaa gcttcgaatt ctgcagtcga 
      301 cggtaccgcg ggcccgggat ccaccggtcg ccaccatgga catcaaggtg gtgttcaccc 
      361 tggtgttcag cgccctggtg caggccaaga gcaccgagtt cgaccccaac atcgacatcg 
      421 tgggcctgga aggcaagttc ggcatcacca acctggaaac cgacctgttc accatctggg 
      481 agaccatgga agtgatgatc aaggccgaca tcgccgacac cgaccgggcc agcaacttcg 
      541 tggccaccga gaccgacgcc aaccggggca agatgcccgg caagaagctg cccctggccg 
      601 tcatcatgga aatggaagcc aacgccttca aggccggctg cacccggggc tgcctgatct 
      661 gcctgagcaa gatcaagtgc accgccaaga tgaaggtgta catccccggc aggtgccacg 
      721 actacggcgg cgacaagaaa accggccagg ccggcatcgt gggcgccatc gtggacatcc 
      781 ccgagatcag cggcttcaaa gaaatggccc ccatggaaca gttcatcgcc caggtggaca 
      841 gatgcgccag ctgcaccacc ggctgcctga agggcctggc caacgtgaag tgcagcgagc 
      901 tgctgaagaa gtggctgccc gaccgctgcg ccagcttcgc cgacaagatc cagaaagagg 
      961 tgcacaacat caagggcatg gccggcgaca ggtgagcggc cgcgactcta gatcataatc 
     1021 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg 
     1081 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat 
     1141 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat 
     1201 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa 
     1261 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc 
     1321 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt 
     1381 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa 
     1441 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg 
     1501 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg 
     1561 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc 
     1621 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa 
     1681 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat 
     1741 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata 
     1801 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat 
     1861 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 
     1921 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 
     1981 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 
     2041 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 
     2101 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga 
     2161 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt 
     2221 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct 
     2281 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct 
     2341 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga 
     2401 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc 
     2461 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg 
     2521 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc 
     2581 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca 
     2641 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga 
     2701 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc 
     2761 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga 
     2821 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca 
     2881 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg 
     2941 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct 
     3001 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc 
     3061 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga 
     3121 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc 
     3181 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc 
     3241 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat 
     3301 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg 
     3361 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc 
     3421 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat 
     3481 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt 
     3541 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac 
     3601 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc 
     3661 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca 
     3721 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta 
     3781 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct 
     3841 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg 
     3901 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc 
     3961 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta 
     4021 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg 
     4081 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt 
     4141 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg 
     4201 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg 
     4261 ccttttgctc acatgtaata aaatatcttt attttcatta catctgtgtg ttggtttttt 
     4321 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact 
     4381 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct acatgttctt 
     4441 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcat 
//

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