pProLabel-N

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-N
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: 4167 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(C ter)
载体抗性: 卡那霉素
筛选标记: Neomycin_1.html' target='_blank'>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: 631628
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-N载体图谱



pProLabel-N 多克隆位点

载体简介

pProLabel-N载体描述

pProLabel-N is a mammalian expression vector designed to express a protein of interest fused to the the n-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.

Propagation in E. coli
 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-N   4167 bp    DNA   circular             9-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631628
FEATURES             Location/Qualifiers
     promoter        1..589
                     /label=CMV\IE\Promoter
     rep_origin      1083..1538
                     /label=f1\origin
     rep_origin      1879..2014
                     /label=SV40\origin
     CDS             2063..2857
                     /gene="Kanamycin/neomycin resistance gene Neomycin phosphotransferase coding sequences"
                     /label=Kan(R)/Neo(R)
                     /note="CDS(Kanamycin/neomycin resistance gene Neomycin phosphotransferase coding sequences)_2"
     polyA_signal    3093..3098
                     /label=HSV\TK\polyA\signal
                     /note="Herpes simplex virus (HSV) thymidine kinase (TK) polyA signal"
     rep_origin      3442..4085
                     /label=pUC\origin
     CDS             666..833
                     /label=ProLabel\tag
     misc_feature    591..665
                     /label=MCS
     polyA_signal    986..1036
                     /label=SV40\polyA\signal
BASE COUNT     1001 a      1093 c      1079 g       994 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 atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg 
      661 gatccagctc caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg 
      721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag 
      781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa taggcggccg 
      841 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac 
      901 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg 
      961 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa 
     1021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc 
     1081 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc 
     1141 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga 
     1201 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc 
     1261 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc 
     1321 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag 
     1381 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa 
     1441 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac 
     1501 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa 
     1561 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 
     1621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga 
     1681 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca 
     1741 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca 
     1801 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc 
     1861 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc 
     1921 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta 
     1981 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga 
     2041 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc 
     2101 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga 
     2161 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct 
     2221 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac 
     2281 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct 
     2341 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt 
     2401 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt 
     2461 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt 
     2521 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag 
     2581 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt 
     2641 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg 
     2701 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg 
     2761 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg 
     2821 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg 
     2881 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat 
     2941 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg 
     3001 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag 
     3061 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt 
     3121 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc 
     3181 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc 
     3241 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata 
     3301 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg 
     3361 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg 
     3421 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt 
     3481 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg 
     3541 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata 
     3601 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca 
     3661 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag 
     3721 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc 
     3781 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga 
     3841 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg 
     3901 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac 
     3961 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg 
     4021 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg 
     4081 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct 
     4141 gtggataacc gtattaccgc catgcat 
//

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