pNFκB-MetLuc2-Reporter

价格:2575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: pNFκB-MetLuc2-Reporter
质粒类型: 哺乳动物细胞表达载体;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: TA
载体大小: 4366 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: 卡那霉素
筛选标记: 新霉素(Neomycin)
克隆菌株: Stbl3
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 哺乳动物表达载体pNFκB-MetLuc2-Reporter是荧光素酶报告载体;
MetLuc是已知分子量最小的荧光素酶;
启动子上游有NFκB增强子元件。
产品目录号: 631743
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

pNFκB-MetLuc2-Reporter载体图谱



pNFκB-MetLuc2-Reporter 载体特征

载体简介

pNFκB-MetLuc2-Reporter载体描述

pNFκB-MetLuc2-Reporter allows you to monitor the activation of the NFκB signal transduction pathway (1–4) directly from the cell culture medium, without cell lysis. The vector contains an NFκB enhancer element (composed of four tandem copies of the NFκB consensus binding sequence; 5), located upstream of the minimal TA promoter (PTA), which consists of the TATA box from the herpes simplex virus thymidine kinase (HSV-TK) promoter. Located downstream of PTA 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 to the NFκB enhancer element allows Metridia luciferase (MetLuc) to be expressed and secreted into the surrounding medium (6).
To prevent read-through transcription of the Metridia luciferase gene, a synthetic transcription
blocker (TB; 7, 8), composed of adjacent polyadenylation and transcription pause sites, is located upstream of the cis-acting enhancer element. 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.

pNFκB-MetLuc2-Reporter is available as part of the Ready-To-Glow? NFκB Secreted Luciferase Reporter
System (Cat. No. 631743). In mammalian cells containing pNFκB-MetLuc2-Reporter, the addition of TNFα, IL-1, or stimulants of lymphokine receptors to the culture medium induces transcription factors to bind to the NFκB enhancer element, thereby initiating transcription of the secreted luciferase reporter gene.
Activation of the NFκB signal transduction pathway can be monitored simply by sampling the culture medium. To monitor the effect of a specific protein on the pathway, pNFκB-MetLuc2-Reporter can be cotransfected with an expression vector containing the gene of interest. pNFkB-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 and 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. NF-κB Transcription Reporter Vectors (July 1998) Clontechniques XIII(3):24–25.
2. Baeuerle, P. A. & Baltimore, D. (1996) Cell 87(1):13–20
3. Baeuerle, P. A. (1998) Curr. Biol. 8(1):R19–R22.
4. Peltz, G. (1997) Curr. Opin. Biotechnol. 8(4):467–473.
5. Pessara, U. & Koch, N. (1990) Mol. Cell Biol. 10(8):4146–4154.
6. Markova, S. V., et al. (2004) J. Biol. Chem. 279(5):3212–3217..
7. Eggermont, J. & Proudfoot, N. (1993) EMBO J. 12(6):2539–2548.
8. Enriquez-Harris, P., et al. (1991) EMBO J. 10(7):1833–1842.


载体序列

LOCUS       pNFkB-MetLuc2-Reporter   4366 bp    DNA   circular             8-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631742,631743
FEATURES             Location/Qualifiers
     enhancer        46..85
                     /label=NFkB\enhancer\element
     promoter        105..111
                     /label=TA\minimal\promoter
     CDS             213..869
                     /label=Metridia\secreted\luciferase
     CDS             2102..2896
                     /label=Kan(R)/Neo(R)
     misc_feature    4154..4307
                     /label=Transcription\blocker
     polyA_signal    1025..1030
                     /label=SV40\early\polyA\signal
     rep_origin      complement(1122..1577)
                     /label=f1\origin
     rep_origin      1918..2056
                     /label=SV40\origin
     rep_origin      3481..4124
                     /label=pUC\origin
BASE COUNT     1064 a      1162 c      1159 g       981 t 
ORIGIN
        1 tagttattac tagcgctacc ggactcagat ctcgagctca agcttgggaa tttccgggaa 
       61 tttccgggaa tttccgggaa tttccagatc tagactctag agggtatata atggaagctc 
      121 gaattccagc ttggcattcc ggtactgttg gtaaaaagct tcgaattctg cagtcgacgg 
      181 taccgcgggc ccgggatcca ccggtcgcca ccatggacat caaggtggtg ttcaccctgg 
      241 tgttcagcgc cctggtgcag gccaagagca ccgagttcga ccccaacatc gacatcgtgg 
      301 gcctggaagg caagttcggc atcaccaacc tggaaaccga cctgttcacc atctgggaga 
      361 ccatggaagt gatgatcaag gccgacatcg ccgacaccga ccgggccagc aacttcgtgg 
      421 ccaccgagac cgacgccaac cggggcaaga tgcccggcaa gaagctgccc ctggccgtca 
      481 tcatggaaat ggaagccaac gccttcaagg ccggctgcac ccggggctgc ctgatctgcc 
      541 tgagcaagat caagtgcacc gccaagatga aggtgtacat ccccggcagg tgccacgact 
      601 acggcggcga caagaaaacc ggccaggccg gcatcgtggg cgccatcgtg gacatccccg 
      661 agatcagcgg cttcaaagaa atggccccca tggaacagtt catcgcccag gtggacagat 
      721 gcgccagctg caccaccggc tgcctgaagg gcctggccaa cgtgaagtgc agcgagctgc 
      781 tgaagaagtg gctgcccgac cgctgcgcca gcttcgccga caagatccag aaagaggtgc 
      841 acaacatcaa gggcatggcc ggcgacaggt gagcggccgc gactctagat cataatcagc 
      901 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac 
      961 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt 
     1021 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct 
     1081 agttgtggtt tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat 
     1141 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 
     1201 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 
     1261 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 
     1321 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 
     1381 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 
     1441 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 
     1501 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 
     1561 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg 
     1621 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat 
     1681 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg 
     1741 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg 
     1801 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt 
     1861 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc 
     1921 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt 
     1981 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc 
     2041 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg 
     2101 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt 
     2161 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc 
     2221 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact 
     2281 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt 
     2341 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca 
     2401 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat 
     2461 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg 
     2521 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga 
     2581 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga 
     2641 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa 
     2701 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga 
     2761 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt 
     2821 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct 
     2881 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac 
     2941 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc 
     3001 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc 
     3061 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc 
     3121 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa 
     3181 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc 
     3241 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca 
     3301 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata 
     3361 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt 
     3421 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc 
     3481 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg 
     3541 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact 
     3601 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg 
     3661 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg 
     3721 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac 
     3781 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca 
     3841 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga 
     3901 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc 
     3961 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct 
     4021 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg 
     4081 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct 
     4141 tttgctcaca tgtaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg 
     4201 tgaatcgata gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc 
     4261 aaaataggct gtccccagtg caagtgcagg tgccagaaca tttctctaca tgttctttcc 
     4321 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat 
//

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