pAcGFP1-C

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-C
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: 4722 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-C是Match-Maker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-C载体图谱



pAcGFP1-C 多克隆位点

pAcGFP1-C 载体特征

载体简介

pAcGFP1-C载体描述

protein (GFP) from Aequorea coerulescens. The fluorescent protein coding sequence in this construct has been human-codon-optimized for efficient expression and enhanced brightness. AcGFP1 protein has an excitation maximum at 475 nm and an emission maximum at 505 nm. Sequences flanking AcGFP1 have been converted to a Kozak consensus translation initiation site (1) to further increase translation efficiency in eukaryotic cells. A gene of interest can be added in-frame downstream of the AcGFP1 coding sequence and expressed as a fusion protein to the C-terminus of AcGFP1. SV40 polyadenylation signals downstream of the AcGFP1 gene direct proper processing of the 3’end of the mRNA transcript. In addition, the vector also contains a SV40 origin of replication in mammalian cells expressing the SV40 T-antigen. A neomycin resistance cassette (Neor) containing an SV40 early promoter, the neomycin/ kanamycin resistance gene of Tn5, and polyadenylation signals from the Herpes simplex virus thymidine kinase (HSV TK) gene, allows selection of stable transformants in eukaryotic cells using G418. A bacterial promoter upstream of the gene expresses kanamycin resistance in E. coli. pAcGFP-C Vector also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.

Provided with the Matchmaker Chemiluminescent CoIP Vector Set, the pAcGFP1-C Vector allows the generation of a N-terminal AcGFP1-bait fusion. Any bait sequence in our current Matchmaker Yeast Two Hybrid System 3 pGBKT7 bait vector can be PCR-amplified using the AcGFP BD FWD/REV universal In-Fusion primer set, and directionally and efficiently cloned into the SalI/HindIII sites of the pAcGFP1-C using our In-Fusion Dry-Down PCR Cloning Kit. The recombinant AcGFP1 vector can be transfected into mammalian cells using any standard transfection method. The resulting expressed AcGFP1- bait fusion protein will automatically be in-frame and can be used to monitor transfection efficiency as well as AcGFP1-bait fusion expression and localization in vivo since the AcGFP1-bait fusion retains the fluorescent properties of the native AcGFP1 protein.

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. Kozak, M. (1987) Nucleic Acids Res. 15(20):8125–8148.
2. 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-C   4722 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    1541..1591
                     /label=SV40\early\polyA\signals
     misc_feature    1638..2093
                     /label=f1\origin
     misc_feature    2434..2569
                     /label=SV40\origin
     misc_feature    2618..3412
                     /label=Kan(R)/Neo(R)
     misc_feature    3648..3666
                     /label=HSV\TK\polyA\signals
     misc_feature    3997..4640
                     /label=pUC\origin
     misc_feature    1348..1408
                     /label=MCS
BASE COUNT     1140 a      1265 c      1232 g      1085 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 gaccagatct ctaagcttgc gaattctggc 
     1381 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta 
     1441 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt 
     1501 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca 
     1561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc 
     1621 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt 
     1681 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc 
     1741 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt 
     1801 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact 
     1861 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg 
     1921 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag 
     1981 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac 
     2041 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg 
     2101 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa 
     2161 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa 
     2221 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc 
     2281 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg 
     2341 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag 
     2401 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc 
     2461 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc 
     2521 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc 
     2581 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca 
     2641 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac 
     2701 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt 
     2761 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc 
     2821 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg 
     2881 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc 
     2941 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc 
     3001 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat 
     3061 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc 
     3121 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca 
     3181 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga 
     3241 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat 
     3301 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc 
     3361 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact 
     3421 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc 
     3481 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg 
     3541 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact 
     3601 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag 
     3661 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg 
     3721 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt 
     3781 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg 
     3841 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat 
     3901 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct 
     3961 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct 
     4021 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca 
     4081 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc 
     4141 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc 
     4201 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct 
     4261 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag 
     4321 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc 
     4381 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg 
     4441 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag 
     4501 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt 
     4561 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac 
     4621 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg 
     4681 ttatcccctg attctgtgga taaccgtatt accgccatgc at 
//

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