pAcGFP1-Golgi

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-Golgi
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: 4.9 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: 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-Golgi组成型表达AcGFP蛋白;含有高尔基体定位信号,是亚细胞定位载体。
产品目录号: 632464
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-Golgi载体图谱



pAcGFP1-Golgi 载体特征

载体简介


pAcGFP1-Golgi Vector encodes a fusion protein consisting of the N-terminal 81 amino acids of the human beta 1,4-galactosyltransferase (GT; 1) directly upstream of the green fluorescent protein (GFP) from Aequorea coerulescens (AcGFP1). The human beta 1,4-galactosyltransferase sequence contains a membrane-anchoring signal peptide that targets the protein to the transmedial region of the Golgi aparatus (2–4). AcGFP1 (excitation maximum = 475 nm; emission maximum = 505 nm) 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. SV40 polyadenylation signals downstream of the protein coding region direct proper processing of the 3' end of the mRNA. The vector contains an SV40 origin for replication and a neomycin resistance (Neor) gene for selection (using G418) in eukaryotic cells (5). A bacterial promoter (P) upstream of Neor expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.


pAcGFP1-Golgi Vector is designed for fluorescent labeling of the Golgi apparatus. pAcGFP1-Golgi can be introduced into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (5). pAcGFP1-Golgi is not intended as a cloning vector; however, the backbone does contain unique restriction sites upstream and downstream of the AcGFP1-Golgi sequence which permit its excision.

Propagation in E. coli
 Suitable host strains: Fusion-Blue competent cells (Cat. No. 636700), DH5α, HB101, and other general purpose strains. Single-stranded DNA production requires a host such as JM109 or XL1-Blue that contains an
F plasmid.
 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-Golgi   4911 bp    DNA   circular             9-DEC-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 632464
FEATURES             Location/Qualifiers
     promoter        1..589
                     /label=CMV\IE\Promoter
     CDS             597..842
                     /label=Human-GT\precursor
     CDS             858..1568
                     /label=AcGFP1
     misc_feature    2801..3595
                     /label=Kan(R)/Neo(R)
     misc_feature    1724..1758
                     /label=SV40\early\polyA\signals
     misc_feature    1821..2276
                     /label=f1\origin
     misc_feature    2338..2366
                     /label=Bacterial\Promoter\for\Kan(R)
     misc_feature    2617..2752
                     /label=SV40\origin
     misc_feature    2450..2679
                     /label=SV40\early\Promoter/Enhancer
     misc_feature    3831..3849
                     /label=HSV\TK\polyA\signals
     misc_feature    4180..4823
                     /label=pUC\origin
BASE COUNT     1157 a      1341 c      1305 g      1108 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 gctagcatga 
      601 ggcttcggga gccgctcctg agcggcagcg ccgcgatgcc aggcgcgtcc ctacagcggg 
      661 cctgccgcct gctcgtggcc gtctgcgctc tgcaccttgg cgtcaccctc gtttactacc 
      721 tggctggccg cgacctgagc cgcctgcccc aactggtcgg agtctccaca ccgctgcagg 
      781 gcggctcgaa cagtgccgcc gccatcgggc agtcctccgg ggagctccgg accggagggg 
      841 ccaaggatcc accggtcatg gtgagcaagg gcgccgagct gttcaccggc atcgtgccca 
      901 tcctgatcga gctgaatggc gatgtgaatg gccacaagtt cagcgtgagc ggcgagggcg 
      961 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc 
     1021 ctgtgccctg gcccaccctg gtgaccaccc tgagctacgg cgtgcagtgc ttctcacgct 
     1081 accccgatca catgaagcag cacgacttct tcaagagcgc catgcctgag ggctacatcc 
     1141 aggagcgcac catcttcttc gaggatgacg gcaactacaa gtcgcgcgcc gaggtgaagt 
     1201 tcgagggcga taccctggtg aatcgcatcg agctgaccgg caccgatttc aaggaggatg 
     1261 gcaacatcct gggcaataag atggagtaca actacaacgc ccacaatgtg tacatcatga 
     1321 ccgacaaggc caagaatggc atcaaggtga acttcaagat ccgccacaac atcgaggatg 
     1381 gcagcgtgca gctggccgac cactaccagc agaatacccc catcggcgat ggccctgtgc 
     1441 tgctgcccga taaccactac ctgtccaccc agagcgccct gtccaaggac cccaacgaga 
     1501 agcgcgatca catgatctac ttcggcttcg tgaccgccgc cgccatcacc cacggcatgg 
     1561 atgagctgta caagtgagcg gccgcgactc tagatcataa tcagccatac cacatttgta 
     1621 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 
     1681 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 
     1741 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 
     1801 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc 
     1861 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 
     1921 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 
     1981 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 
     2041 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 
     2101 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 
     2161 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 
     2221 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 
     2281 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 
     2341 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 
     2401 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 
     2461 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 
     2521 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 
     2581 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 
     2641 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 
     2701 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 
     2761 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga 
     2821 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc 
     2881 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc 
     2941 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc 
     3001 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac 
     3061 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc 
     3121 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac 
     3181 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg 
     3241 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct 
     3301 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt 
     3361 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg 
     3421 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac 
     3481 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg 
     3541 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg 
     3601 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat 
     3661 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc 
     3721 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg 
     3781 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata 
     3841 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc 
     3901 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt 
     3961 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac 
     4021 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta 
     4081 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc 
     4141 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa 
     4201 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca 
     4261 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta 
     4321 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc 
     4381 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca 
     4441 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta 
     4501 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag 
     4561 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt 
     4621 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc 
     4681 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac 
     4741 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac 
     4801 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc 
     4861 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t 
//

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