pProLabel-C

价格:2000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: pProLabel-C
质粒类型: 哺乳动物表达载体
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: 4161 bp
5' 测序引物及序列: CMV Forward: CGCAAATGGGCGGTAGGCGTG
3' 测序引物及序列: --
载体标签: ProLabel tag(N- ter)
载体抗性: Kanamycin.html' target='_blank'>卡那霉素
筛选标记: 新霉素(Neomycin)
克隆菌株: DH5α 等
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: ProLabel tag 是一个分子量约6KD的标签,检测的灵敏度高。
产品目录号: 630458
稳定性: 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

pProLabel-C载体图谱



pProLabel-C 多克隆位

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pProLabel-C 载体特征

载体简介

pProLabel-C载体描述

pProLabel-C is a mammalian expression vector designed to express a protein of interest fused to the the C-terminus of a 6 kDa ProLabel tag. The resulting fusion protein can be used in a variety of functional assays and quantitated with our ProLabel Detection Kit II (Cat. No. 631629). The Detection Kit provides all of the components needed to perform enzyme fragment complementation assays (1, 2). In these assays, two inactive enzyme fragments (the ProLabel tag, and a larger Enzyme Acceptor) are combined to form a complete, active enzyme that cleaves the Galacton Star chemiluminescent substrate. The resulting signal can be detected and quantitated with any standard luminometer.
The pProLabel-C vector contains a CMV promoter that drives strong, constitutive expression of the fusion protein, and an SV40 polyadenylation signal that directs processing of the 3’end of the mRNA transcript. The vector also contains a kanamycin/neomycin resistance cassette (Kanr/Neor) that allows G418 selection of stably transfected eukaryotic cells; a bacterial promoter upstream of this cassette allows kanamycin or neomycin selection of transformed bacterial cells. In addition, pProLabel-C contains an SV40 origin of replication for propagation in mammalian cells that express SV40 T-antigen, a pUC origin for propagation in E. coli and an f1 origin for the production of single-stranded DNA.

In order to create a fusion of your protein of interest and the ProLabel tag, your gene of interest must be in the same reading frame as the ProLabel tag sequence, and there can be no intervening stop codons. Prolabel vector constructs can be transfected into mammalian cells using standard transfection methods. ProLabel fusion protein expression levels can be measured quantitatively from mammalian cell lysates using the method described in the ProLabel Detection Kit II Protocol-at-a-Glance (PT3987-2). This highly sensitive and rapid procedure obviates the need for Western analysis. The assay is designed to be used with any ProLabel fusion in a variety of functional assays—from measuring target gene expression in RNAi knockdown studies to measuring protein interactions in coimmunoprecipitation studies.

质粒扩增方法
 Suitable host strains: DH5α, Fusion Blue, and other general purpose strains. Single-stranded DNA production requires a host containing an F plasmid, such as JM101 or XL1-Blue.
 Selectable marker: plasmid confers resistance to kanamycin (50 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC
 Copy number: ~500
 Plasmid incompatibility group: pMB1/ColE1

参考文献
1. Gorman, C. (1985) In DNA cloning: A practical approach, Vol. II. Ed. D.M. Glover. (IRL Press, Oxford, UK) pp. 143–190.
2. Eglen, R.M. and Singh, R. (2002) Comb. Chem. High Throughput Screen. 6: 381–387.

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.

载体序列

LOCUS       pProLabel-C   4161 bp    DNA   circular             1-FEB-2010
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 630458,630461,630460,631628
FEATURES             Location/Qualifiers
     misc_feature    1..589
                     /label=CMV\IE\Promoter
     misc_feature    613..780
                     /label=ProLabel\tag
     misc_feature    782..825
                     /label=MCS
     misc_feature    980..1030
                     /label=SV40\polyA\signals
     misc_feature    1077..1532
                     /label=f1\origin
     misc_feature    1873..2008
                     /label=SV40\origin
     misc_feature    2057..2851
                     /label=Kan(R)/Neo(R)
     misc_feature    3087..3105
                     /label=HSV\TK\polyA\signals
     misc_feature    3436..4079
                     /label=pUC\origin
BASE COUNT     1002 a      1092 c      1074 g       993 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 ccggtcgcca ccatgagctc caattcactg gccgtcgttt tacaacgtcg tgactgggaa 
      661 aaccctggcg ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt 
      721 aatagcgaag aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa 
      781 caagcttcga attctgcagt cgacggtacc gcgggcccgg gatccaccgg atctagataa 
      841 ctgatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca 
      901 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt 
      961 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt 
     1021 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta acgcgtaaat 
     1081 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 
     1141 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 
     1201 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 
     1261 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 
     1321 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 
     1381 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 
     1441 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 
     1501 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg 
     1561 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca 
     1621 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac 
     1681 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga 
     1741 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc 
     1801 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc 
     1861 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc 
     1921 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag 
     1981 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg 
     2041 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg 
     2101 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc 
     2161 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg 
     2221 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt 
     2281 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg 
     2341 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat 
     2401 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca 
     2461 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca 
     2521 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa 
     2581 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa 
     2641 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc 
     2701 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga 
     2761 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc 
     2821 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac 
     2881 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg 
     2941 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 
     3001 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata 
     3061 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg 
     3121 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg 
     3181 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt 
     3241 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca 
     3301 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag 
     3361 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac 
     3421 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc 
     3481 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat 
     3541 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat 
     3601 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct 
     3661 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt 
     3721 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg 
     3781 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta 
     3841 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg 
     3901 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg 
     3961 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc 
     4021 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg 
     4081 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat 
     4141 aaccgtatta ccgccatgca t 
//

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