pAcGFP1-p53

价格:2500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: pAcGFP1-p53
质粒类型: 哺乳动物细胞表达载体;荧光报告载体
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 IE
载体大小: 5932bp
5' 测序引物及序列: --
3' 测序引物及序列: --
载体标签: AcGFP1 (N-端)
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, HB101
宿主细胞(系): 常规细胞系(293、CV-1、CHO等)
备注: 哺乳动物载体pAcGFP1-p53组成型表达N端带有AcGFP标签的P53蛋白;
AcGFP1与GFP相比,密码子经过了优化,更适合在哺乳动物细胞中高水平表达,同时也增强了亮度。
产品目录号: 630458
稳定性: 稳表达 或 瞬表达
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

pAcGFP1-p53载体图谱



pAcGFP1-p53 载体特征

载体简介

pAcGFP1-p53载体描述

pAcGFP1-p53 Vector encodes a green fluorescent protein (GFP) from Aequorea coerulescens (excitation maximum = 475 nm; emission maximum = 505 nm) and the gene encoding the human tumor suppressor p53 (ref.). SV40 polyadenylation signals downstream of the AcGFP1- p53 fusion direct proper processing of the 3’end of the AcGFP1-p53 mRNA. AcGFP1 contains silent mutations that create an open reading frame comprised almost entirely of optimized human codons. These changes increase the translational efficiency of the AcGFP1 mRNA and consequently the expression of AcGFP1 in mammalian and plant cells. The vector backbone also contains an SV40 origin for replication in any mammalian cell line that expresses the SV40 T antigen. A neomycin resistance cassette (Neor), consisting of the SV40 early promoter, the neomycin/kanamycin resistance gene of Tn5, and polyadenylation signals from the herpes simplex virus thymidine kinase (HSV-TK) gene, allows stably transfected eukaryotic cells to be selected using G418. A bacterial promoter upstream of this cassette drives expression of the gene encoding kanamycin resistance in E. coli. The pAcGFP1-p53 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

The pAcGFP1-p53 is provided in the Matchmaker Chemiluminescent Co-IP Vector Set as a positive bait control construct. In mammalian cells, this vector expresses the AcGFP1-p53 fusion protein—AcGFP1 serves as a tag, through which a polyclonal antibody against AcGFP1 can be used for immunoprecipitation, while p53 serves as the bait for the capture of interacting proteins via coimmunoprecipitation. Because p53 interacts with SV40 large T (1), in lysates of cells cotransfected with pAcGFP1-p53 and ProLabel-T, there is a high level of ProLabel activity detected in the Co-IP of this positive control for interacting pairs of proteins. The fluorescence from AcGFP1-p53 fusion protein also allows for easy detection of fusion expression and localization in vivo, as well as offering a mode for determining transfection efficiency.

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 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

载体序列

LOCUS       pAcGFP1-p53   5932 bp    DNA   circular             1-FEB-2010
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 630458
FEATURES             Location/Qualifiers
     misc_feature    1..589
                     /label=CMV\IE\Promoter
     misc_feature    613..1329
                     /label=AcGFP1
     misc_feature    1417..2376
                     /label=p53\(truncated)
     misc_feature    2751..2801
                     /label=SV40\early\polyA\signals
     misc_feature    2848..3303
                     /label=f1\origin
     misc_feature    3644..3779
                     /label=SV40\origin
     misc_feature    3828..4622
                     /label=Kan(R)/Neo(R)
     misc_feature    4858..4876
                     /label=HSV\TK\polyA\signals
     misc_feature    5207..5850
                     /label=pUC\origin
BASE COUNT     1436 a      1610 c      1543 g      1343 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 ccatggtgag caagggcgcc gagctgttca ccggcatcgt gcccatcctg 
      661 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc 
      721 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg 
      781 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc 
      841 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag 
      901 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag 
      961 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac 
     1021 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac 
     1081 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc 
     1141 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg 
     1201 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc 
     1261 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag 
     1321 ctgtacaagt ccggacttaa ggcctctgtc gacgccgccg aggagcagaa gctgatctca 
     1381 gaggaggacc tgcatatggc catggaggcc gaattccctg tcaccgagac ccctgggcca 
     1441 gtggcccctg ccccagccac tccatggccc ctgtcatctt ttgtcccttc tcaaaaaact 
     1501 taccagggca actatggctt ccacctgggc ttcctgcagt ctgggacagc caagtctgtt 
     1561 atgtgcacgt actctcctcc cctcaataag ctattctgcc agctggcgaa gacgtgccct 
     1621 gtgcagttgt gggtcagcgc cacacctcca gctgggagcc gtgtccgcgc catggccatc 
     1681 tacaagaagt cacagcacat gacggaggtc gtgagacgct gcccccacca tgagcgctgc 
     1741 tccgatggtg atggcctggc tcctccccag catcttatcc gggtggaagg aaatttgtat 
     1801 cccgagtatc tggaagacag gcagactttt cgccacagcg tggtggtacc ttatgagcca 
     1861 cccgaggccg gctctgagta taccaccatc cactacaagt acatgtgtaa tagctcctgc 
     1921 atggggggca tgaaccgccg acctatcctt accatcatca cactggaaga ctccagtggg 
     1981 aaccttctgg gacgggacag ctttgaggtt cgtgtttgtg cctgccctgg gagagaccgc 
     2041 cgtacagaag aagaaaattt ccgcaaaaag gaagtccttt gccctgaact gcccccaggg 
     2101 agcgcaaaga gagcgctgcc cacctgcaca agcgcctctc ccccgcaaaa gaaaaaacca 
     2161 cttgatggag agtatttcac cctcaagatc cgcgggcgta aacgcttcga gatgttccgg 
     2221 gagctgaatg aggccttaga gttaaaggat gcccatgcta cagaggagtc tggagacagc 
     2281 agggctcact ccagctacct gaagaccaag aagggccagt ctacttcccg ccataaaaaa 
     2341 acaatggtca agaaagtggg gcctgactca gactgactgc ctctgcatcc cggatccgtc 
     2401 gacctgcagc ggccgcataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg 
     2461 gttttttgcg cgcttgcagc caagctaatt ccgggcgaat ttcttatgat ttatgatttt 
     2521 tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg acaagcttgc 
     2581 gaattctggc ggatccaccg gatctagata actgatcata atcagccata ccacatttgt 
     2641 agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat 
     2701 gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa 
     2761 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 
     2821 caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg 
     2881 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc 
     2941 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga 
     3001 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg 
     3061 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag 
     3121 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga 
     3181 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg 
     3241 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg 
     3301 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa 
     3361 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt 
     3421 gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg 
     3481 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc 
     3541 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca 
     3601 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc 
     3661 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc 
     3721 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct 
     3781 aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag 
     3841 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg 
     3901 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc 
     3961 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag 
     4021 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca 
     4081 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat 
     4141 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata 
     4201 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac 
     4261 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc 
     4321 tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg 
     4381 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg 
     4441 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta 
     4501 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg 
     4561 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct 
     4621 gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga 
     4681 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc 
     4741 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg 
     4801 gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat 
     4861 aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc 
     4921 ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt 
     4981 ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa 
     5041 cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt 
     5101 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac 
     5161 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa 
     5221 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc 
     5281 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt 
     5341 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg 
     5401 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc 
     5461 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt 
     5521 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga 
     5581 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct 
     5641 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg 
     5701 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca 
     5761 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa 
     5821 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt 
     5881 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc at 
//

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