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 |
买家导航
pAcGFP1-Hyg-C1载体质粒基本信息
pAcGFP1-Hyg-C1质粒图谱载体图谱和pAcGFP1-Hyg-C1载体序列质粒序列多克隆位点信息
pAcGFP1-Hyg-C1质粒载体简介
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.
pAcGFP1-Hyg-C1质粒序列载体序列
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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