| 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 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
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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