pAcGFP1-Lam

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-Lam
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: 5462 bp
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-Lam组成型表达N端AcGFP融合蛋白LaminA/C;
pAcGFP1-Lam是MatchMaker系统载体;
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-Lam载体图谱



pAcGFP1-Lam 载体特征

载体简介

pAcGFP1-Lam载体描述

pAcGFP1-Lamin encodes a green fluorescent protein (GFP) from Aequorea coerulescens (excitation maximum = 475 nm; emission maximum = 505 nm) and the gene encoding the human nuclear lamin (1). SV40 polyadenylation signals downstream of the AcGFP1-Lamin fusion direct proper processing of the 3’end of the AcGFP1-Lamin 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-Lamin backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

The pAcGFP1-Lamin is provided in the Matchmaker Chemiluminescent Co-IP Vector Set (Cat. No. 630458) as a negative bait control construct. In mammalian cells, this vector expresses the AcGFP1-Lamin fusion protein—the AcGFP1 serves as a tag, through which a polyclonal antibody against AcGFP1 can be used for immunoprecipitation, while lamin serves as the bait for the capture of interacting proteins via coimmunoprecipitation. Because lamin does not interact with SV40 large T antigen (2), in lysates of cells cotransfected with pAcGFP1-Lamin and pProLabel-T, there is no ProLabel activity detected in the Co-IP of this control for noninteracting pairs of proteins. The fluorescence from AcGFP1-Lamin 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

References
1. Sturman, N. et al. (1998) J. Stuct. Biol. 122(1–2) 42–66.
2. Matchmaker Co-IP Kit (April 1999) Clontechniques XIV(2): 14–15.
3. Gorman, C. (1985) In DNA Cloning: A Practical Approach, Vol. II. Ed. D.M. Glover. (IRL Press, Oxford, UK) pp. 143–190.


载体序列

LOCUS       pAcGFP1-Lam   5462 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    1426..1920
                     /label=Lamin\A/C
     misc_feature    2281..2331
                     /label=SV40\early\poly\signals
     misc_feature    2378..2833
                     /label=f1\origin
     misc_feature    3174..3309
                     /label=SV40\origin
     misc_feature    3358..4152
                     /label=Kan(R)/Neo(R)
     misc_feature    4388..4406
                     /label=HSV\TK\polyA\signals
     misc_feature    4737..5380
                     /label=pUC\origin
BASE COUNT     1328 a      1439 c      1472 g      1223 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 gaattcccgg ggatctctga agaggtggtc 
     1441 agccgcgagg tgtccggcat caaggccgcc tacgaggccg agctcgggga tgcccgcaag 
     1501 acccttgact cagtagccaa ggagcgcgcc cgcctgcagc tggagctgag caaagtgcgt 
     1561 gaggagttta aggagctgaa agcgcgcaat accaagaagg agggtgacct gatagctgct 
     1621 caggctcggc tgaaggacct ggaggctctg ctgaactcca aggaggccgc actgagcact 
     1681 gctctcagtg agaagcgcac gctggagggc gagctgcatg atctgcgggg ccaggtggcc 
     1741 aagcttgagg cagccctagg tgaggccaag aagcaacttc aggatgagat gctgcggcgg 
     1801 gtggatgctg agaacaggct gcagaccatg aaggaggaac tggacttcca gaagaacatc 
     1861 tacagtgagg agctgcgtga gaccaagcgc cgtcatgaga cccgactggt ggagattgac 
     1921 aggatccgtc gacctgcagc ggccgcataa ctagcataac cccttggggc ctctaaacgg 
     1981 gtcttgaggg gttttttgcg cgcttgcagc caagctaatt ccgggcgaat ttcttatgat 
     2041 ttatgatttt tattattaaa taagttataa aaaaaataag tgtatacaaa ttttaaagtg 
     2101 acaagcttgc gaattctggc ggatccaccg gatctagata actgatcata atcagccata 
     2161 ccacatttgt agaggtttta cttgctttaa aaaacctccc acacctcccc ctgaacctga 
     2221 aacataaaat gaatgcaatt gttgttgtta acttgtttat tgcagcttat aatggttaca 
     2281 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 
     2341 gtggtttgtc caaactcatc aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg 
     2401 ttaaaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc 
     2461 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt 
     2521 tggaacaaga gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc 
     2581 tatcagggcg atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg 
     2641 tgccgtaaag cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga 
     2701 aagccggcga acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg 
     2761 ctggcaagtg tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg 
     2821 ctacagggcg cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta 
     2881 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt 
     2941 caataatatt gaaaaaggaa gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc 
     3001 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc 
     3061 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 
     3121 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 
     3181 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 
     3241 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt 
     3301 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg 
     3361 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc 
     3421 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg 
     3481 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa 
     3541 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc 
     3601 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat 
     3661 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg 
     3721 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc 
     3781 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag 
     3841 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc 
     3901 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc 
     3961 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata 
     4021 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc 
     4081 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac 
     4141 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc 
     4201 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt 
     4261 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc 
     4321 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 
     4381 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg 
     4441 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat 
     4501 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc 
     4561 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt 
     4621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata 
     4681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag 
     4741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa 
     4801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt 
     4861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc 
     4921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa 
     4981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa 
     5041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc 
     5101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa 
     5161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa 
     5221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg 
     5281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc 
     5341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg 
     5401 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc 
     5461 at 
//

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