pAcGFP1-Mem

价格:2300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-Mem
质粒类型: 哺乳动物细胞表达载体;荧光报告载体;亚细胞定位载体
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: 4.8 kb
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: Neomycin_1.html' target='_blank'>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-Mem组成型表达AcGFP,定位于细胞膜。
产品目录号: 632491
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-Mem载体图谱



pAcGFP1-Mem 载体特征1
pAcGFP1-Mem 载体特征2

载体简介

pAcGFP1-Mem载体描述

pAcGFP1-Mem encodes a fusion protein consisting of the N-terminal 20 amino acids of neuromodulin, also called GAP-43 (1), and the Aequorea coerulescens green fluorescent protein AcGFP1. (Excitation maximum = 475 nm; emission maximum = 505 nm.) The coding sequence of the AcGFP1 gene contains silent base changes, which correspond to human codon-usage preferences (2). The neuromodulin fragment contains a signal for posttranslational palmitoylation of cysteines 3 and 4 that targets AcGFP1 to cellular membranes, and the plasma membrane in particular. Expression of AcGFP1-Mem is driven by the immediate early promoter of CMV (PCMV IE). The vector contains an SV40 origin of replication and a neomycin resistance (Neor) gene for selection in mammalian cells. A bacterial promoter upstream of this cassette (P) expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC19 origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

pAcGFP1-Mem can be transfected into mammalian cells using any standard method. If required, stable transformants can be selected using G418 (3). Expression of AcGFP1-Mem in mammalian cells results in strong labeling of the plasma membrane and allows easy tracking of individual cells in a population. This membrane labeling also permits the study of fine cellular processes such as neuronal axons (4), leading edges of migrating cells, filopodia, or microvilli on cell surfaces. pAc- GFP1-Mem cannot be used as an exclusive plasma membrane marker because it also partially labels intracellular membranes.

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: high

References
1. Skene, J. H. P. & Virag, I. (1989) J. Cell. Biol. 108:613–625.
2. Haas, J., et al. (1996) Curr. Biol. 6:315–324.
3. Gorman, C. (1985) In DNA cloning: a practical approach, vol. II. Ed. D. M. Glover. (IRL Press, Oxford, U.K.), pp. 143–190.
4. Moriyoshi, K., et al. (1996) Neuron 16:255–260.

载体序列

LOCUS       pAcGFP1-Mem_Hyg   5846 bp    DNA   circular            10-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 632491
FEATURES             Location/Qualifiers
     CDS             679..738
                     /label=Membrane
                     /note="Neuromodulin GAP-43"
     CDS             739..1449
                     /label=AcGFP1
     promoter        1..589
                     /label=CMV\IE\Promoter
     rep_origin      5121..5764
                     /label=pUC\origin
     rep_origin      complement(1703..2158)
                     /label=f1\origin
     rep_origin      2499..2637
                     /label=SV40\origin
     CDS             4098..4958
                     /label=Amp(R)
     CDS             2656..3681
                     /label=Hyg(R)
     polyA_signal    1606..1611
                     /label=SV40\early\polyA\signal
     polyA_signal    1635..1640
                     /label=SV40\early\polyA\signal
     polyA_signal    3829..3834
                     /label=SV40\early\polyA\signal
     polyA_signal    3858..3863
                     /label=SV40\early\polyA\signal
     misc_feature    2555..2561
                     /label=SV40\early\Promoter
     promoter        4028..4056
                     /label=Amp(R)\Promoter
BASE COUNT     1484 a      1487 c      1484 g      1391 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 gatccaccgg tcgccaccat gctgtgctgt atgagaagaa ccaaacaggt tgaaaagaat 
      721 gatgaggacc aaaagatcag caagggcgcc gagctgttca ccggcatcgt gcccatcctg 
      781 atcgagctga atggcgatgt gaatggccac aagttcagcg tgagcggcga gggcgagggc 
      841 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcctgtg 
      901 ccctggccca ccctggtgac caccctgagc tacggcgtgc agtgcttctc acgctacccc 
      961 gatcacatga agcagcacga cttcttcaag agcgccatgc ctgagggcta catccaggag 
     1021 cgcaccatct tcttcgagga tgacggcaac tacaagtcgc gcgccgaggt gaagttcgag 
     1081 ggcgataccc tggtgaatcg catcgagctg accggcaccg atttcaagga ggatggcaac 
     1141 atcctgggca ataagatgga gtacaactac aacgcccaca atgtgtacat catgaccgac 
     1201 aaggccaaga atggcatcaa ggtgaacttc aagatccgcc acaacatcga ggatggcagc 
     1261 gtgcagctgg ccgaccacta ccagcagaat acccccatcg gcgatggccc tgtgctgctg 
     1321 cccgataacc actacctgtc cacccagagc gccctgtcca aggaccccaa cgagaagcgc 
     1381 gatcacatga tctacttcgg cttcgtgacc gccgccgcca tcacccacgg catggatgag 
     1441 ctgtacaagt gaagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg 
     1501 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg 
     1561 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca 
     1621 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac 
     1681 tcatcaatgt atcttaaggc gtaaattgta agcgttaata ttttgttaaa attcgcgtta 
     1741 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat 
     1801 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca 
     1861 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc 
     1921 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta 
     1981 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg 
     2041 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg 
     2101 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca 
     2161 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat 
     2221 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa 
     2281 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa 
     2341 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 
     2401 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 
     2461 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 
     2521 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 
     2581 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 
     2641 tttgcaaaga tcgatatgaa aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg 
     2701 atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga agaatctcgt 
     2761 gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat 
     2821 ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct cccgattccg 
     2881 gaagtgcttg acattgggga attcagcgag agcctgacct attgcatctc ccgccgtgca 
     2941 cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct gcagccggtc 
     3001 gcggaggcca tggatgcgat cgctgcggcc gatcttagcc agacgagcgg gttcggccca 
     3061 ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg cgcgattgct 
     3121 gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg 
     3181 caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg 
     3241 cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac agcggtcatt 
     3301 gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat cttcttctgg 
     3361 aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag gcatccggag 
     3421 cttgcaggat cgccgcggct ccgggcgtat atgctccgca ttggtcttga ccaactctat 
     3481 cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg atgcgacgca 
     3541 atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc 
     3601 gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg ccccagcact 
     3661 cgtccgaggg caaaggaata gagttctaga ggatcataat cagccatacc acatttgtag 
     3721 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga 
     3781 atgcaattgt tgttgttaac ttgtttattg cagcttgtaa tggttacaaa taaagcaata 
     3841 gcatcacaaa tttcacaaat aaagcatttt tttcactgcc tcgcgatcca caaccatggc 
     3901 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt 
     3961 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 
     4021 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat 
     4081 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg 
     4141 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg 
     4201 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc 
     4261 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat 
     4321 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact 
     4381 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca 
     4441 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact 
     4501 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg 
     4561 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg 
     4621 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg 
     4681 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg 
     4741 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag 
     4801 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc 
     4861 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga 
     4921 tcgctgagat aggtgcctca ctgattaagc attggtaagg cgcgccaggc cctgccatag 
     4981 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga 
     5041 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt 
     5101 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc 
     5161 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc 
     5221 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac 
     5281 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac 
     5341 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt 
     5401 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct 
     5461 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat 
     5521 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt 
     5581 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg 
     5641 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt 
     5701 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt 
     5761 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg 
     5821 tggataaccg tattaccgcc atgcat 
//

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