| 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 |
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-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
//
载体图谱质粒图谱pdf版和相关资料下载
载体质粒应用举例
pAcGFP1-C3
2015-08-16pAcGFP1-C2
2015-08-09pAcGFP1-C1
2015-08-08pAcGFP1-C In-Fusion Ready
2015-08-08
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