| 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
载体基本信息
载体图谱质粒图谱和多克隆位点信息
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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