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: | 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 |
买家导航
pAcGFP1-p53载体质粒基本信息
pAcGFP1-p53质粒图谱载体图谱和pAcGFP1-p53载体序列质粒序列多克隆位点信息
pAcGFP1-p53质粒载体简介
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
pAcGFP1-p53质粒序列载体序列
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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