pAcGFP1-Hyg-C1

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-Hyg-C1
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: 5.8 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: 氨苄青霉素
筛选标记: 潮霉素(Hygromycin)
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-Hyg-C1组成型表达N端AcGFP融合蛋白;
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: 632492
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-Hyg-C1载体图谱



pAcGFP1-Hyg-C1 多克隆位点

pAcGFP1-Hyg-C1 载体特征1
pAcGFP1-Hyg-C1 载体特征2

载体简介

pAcGFP1-Hyg-C1载体描述

pAcGFP1-Hyg-C1 encodes a green fluorescent protein (GFP) from Aequorea coerulescens. (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 (1). The MCS in pAcGFP1-Hyg-C1 is located downstream of the AcGFP1 coding region, allowing the construction of a C-terminal fusion protein with AcGFP1 when genes are cloned in the same reading frame as AcGFP1 and there are no intervening stop codons. SV40 polyadenylation signals downstream of the AcGFP1 gene direct proper processing of the 3' end of the AcGFP1 mRNA. The vector backbonealso contains an SV40 origin for replication in mammalian cells expressing the SV40 T-antigen. A hygromycin resistance cassette (Hygr), consisting of the SV40 early promoter, the hygromycin resistance gene, and SV40 polyadenylation signals, allows stably transfected eukaryotic cells to be selected using hygromycin. A bacterial promoter upstream of the ampicillin gene expresses ampicillin resistance in E. coli. The pAcGFP1-Hyg-C1 backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

Fusions to the C terminus of AcGFP1 retain the fluorescent properties of the native protein, allowing the localization of the fusion protein in vivo. The target gene should be cloned into pAcGFP1-Hyg-C1 so that it is in frame with the AcGFP1 coding sequences, with no intervening in-frame stop codons. The recombinant AcGFP1 vector can be transfected into mammalian cells using any standard transfection method. If required, stable transformants can be selected using hygromycin. pAcGFP1-Hyg-C1 can also be used simply to express AcGFP1 in a cell line of interest (e.g., as a transfection marker).

Propagation in E. coli
 Suitable host strains: DH5α and other general purpose strains.
 Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) in E. coli hosts.
 E. coli replication origin: pUC

Reference
1. Haas, J., et al. (1996) Curr. Biol. 6:315–324.

载体序列

LOCUS       pAcGFP1-Hyg-C1   5790 bp    DNA   circular            11-NOV-2009
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 632492
FEATURES             Location/Qualifiers
     promoter        1..589
                     /label=CMV\IE\Promoter
     CDS             613..1329
                     /label=AcGFP1
     rep_origin      1647..2102
                     /label=f1\origin
     rep_origin      2443..2581
                     /label=SV40\origin
     rep_origin      5065..5708
                     /label=pUC\origin
     CDS             2600..3625
                     /label=Hyg(R)
     CDS             4042..4902
                     /label=Amp(R)
     polyA_signal    1550..1555
                     /label=SV40\early\polyA\signal
     polyA_signal    1579..1584
                     /label=SV40\early\polyA\signal
     polyA_signal    3773..3778
                     /label=SV40\early\polyA\signal
     polyA_signal    3802..3807
                     /label=SV40\early\polyA\signal
     misc_feature    606..616
                     /label=Kozak\sequence
     misc_feature    2276..2505
                     /label=SV40\early\Promoter
BASE COUNT     1462 a      1478 c      1466 g      1384 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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc 
     1381 gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac cacatttgta 
     1441 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg 
     1501 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat 
     1561 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc 
     1621 aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt taaaattcgc 
     1681 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc 
     1741 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag 
     1801 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga 
     1861 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc 
     1921 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa 
     1981 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt 
     2041 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc 
     2101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat 
     2161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg 
     2221 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt 
     2281 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca 
     2341 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat 
     2401 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg 
     2461 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc 
     2521 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta 
     2581 ggcttttgca aagatcgata tgaaaaagcc tgaactcacc gcgacgtctg tcgagaagtt 
     2641 tctgatcgaa aagttcgaca gcgtctccga cctgatgcag ctctcggagg gcgaagaatc 
     2701 tcgtgctttc agcttcgatg taggagggcg tggatatgtc ctgcgggtaa atagctgcgc 
     2761 cgatggtttc tacaaagatc gttatgttta tcggcacttt gcatcggccg cgctcccgat 
     2821 tccggaagtg cttgacattg gggaattcag cgagagcctg acctattgca tctcccgccg 
     2881 tgcacagggt gtcacgttgc aagacctgcc tgaaaccgaa ctgcccgctg ttctgcagcc 
     2941 ggtcgcggag gccatggatg cgatcgctgc ggccgatctt agccagacga gcgggttcgg 
     3001 cccattcgga ccgcaaggaa tcggtcaata cactacatgg cgtgatttca tatgcgcgat 
     3061 tgctgatccc catgtgtatc actggcaaac tgtgatggac gacaccgtca gtgcgtccgt 
     3121 cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac tgccccgaag tccggcacct 
     3181 cgtgcacgcg gatttcggct ccaacaatgt cctgacggac aatggccgca taacagcggt 
     3241 cattgactgg agcgaggcga tgttcgggga ttcccaatac gaggtcgcca acatcttctt 
     3301 ctggaggccg tggttggctt gtatggagca gcagacgcgc tacttcgagc ggaggcatcc 
     3361 ggagcttgca ggatcgccgc ggctccgggc gtatatgctc cgcattggtc ttgaccaact 
     3421 ctatcagagc ttggttgacg gcaatttcga tgatgcagct tgggcgcagg gtcgatgcga 
     3481 cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca caaatcgccc gcagaagcgc 
     3541 ggccgtctgg accgatggct gtgtagaagt actcgccgat agtggaaacc gacgccccag 
     3601 cactcgtccg agggcaaagg aatagagttc tagaggatca taatcagcca taccacattt 
     3661 gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa 
     3721 atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc 
     3781 aatagcatca caaatttcac aaataaagca tttttttcac tgcctcgcga tccacaacca 
     3841 tggcgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt 
     3901 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt 
     3961 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa 
     4021 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt 
     4081 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat 
     4141 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag 
     4201 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg 
     4261 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata 
     4321 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat 
     4381 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc 
     4441 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg 
     4501 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac 
     4561 gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact 
     4621 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa 
     4681 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct 
     4741 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc 
     4801 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga 
     4861 cagatcgctg agataggtgc ctcactgatt aagcattggt aaggcgcgcc aggccctgcc 
     4921 atagcctcag gttactcata tatactttag attgatttaa aacttcattt ttaatttaaa 
     4981 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt 
     5041 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt 
     5101 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt 
     5161 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag 
     5221 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta 
     5281 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat 
     5341 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg 
     5401 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg 
     5461 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac 
     5521 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga 
     5581 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt 
     5641 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta 
     5701 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat 
     5761 tctgtggata accgtattac cgccatgcat 
//

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