| 出品公司: | Clontech |
|---|---|
| 载体名称: | pAcGFP1-Golgi |
| 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: | 4.9 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: | 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-Golgi组成型表达AcGFP蛋白；含有高尔基体定位信号，是亚细胞定位载体。 |
| 产品目录号: | 632464 |
| 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-Golgi载体质粒基本信息
pAcGFP1-Golgi质粒图谱载体图谱和pAcGFP1-Golgi载体序列质粒序列多克隆位点信息
pAcGFP1-Golgi质粒载体简介
pAcGFP1-Golgi Vector encodes a fusion protein consisting of the N-terminal 81 amino acids of the human beta 1,4-galactosyltransferase (GT; 1) directly upstream of the green fluorescent protein (GFP) from Aequorea coerulescens (AcGFP1). The human beta 1,4-galactosyltransferase sequence contains a membrane-anchoring signal peptide that targets the protein to the transmedial region of the Golgi aparatus (2–4). AcGFP1 (excitation maximum = 475 nm; emission maximum = 505 nm) contains silent mutations that create an open reading frame comprised almost entirely of optimized human codons. These changes increase the translational efficiency of the AcGFP1 mRNA and, consequently, the expression of AcGFP1 in mammalian and plant cells. SV40 polyadenylation signals downstream of the protein coding region direct proper processing of the 3' end of the mRNA. The vector contains an SV40 origin for replication and a neomycin resistance (Neor) gene for selection (using G418) in eukaryotic cells (5). A bacterial promoter (P) upstream of Neor expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
pAcGFP1-Golgi Vector is designed for fluorescent labeling of the Golgi apparatus. pAcGFP1-Golgi can be introduced into mammalian cells using any standard transfection method. If required, stable transformants can be selected using G418 (5). pAcGFP1-Golgi is not intended as a cloning vector; however, the backbone does contain unique restriction sites upstream and downstream of the AcGFP1-Golgi sequence which permit its excision.
Propagation in E. coli
Suitable host strains: Fusion-Blue competent cells (Cat. No. 636700), DH5α, HB101, and other general purpose strains. Single-stranded DNA production requires a host such as JM109 or XL1-Blue that contains an
F plasmid.
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
pAcGFP1-Golgi质粒序列载体序列
LOCUS pAcGFP1-Golgi 4911 bp DNA circular 9-DEC-2009
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat. No. 632464
FEATURES Location/Qualifiers
promoter 1..589
/label=CMV\IE\Promoter
CDS 597..842
/label=Human-GT\precursor
CDS 858..1568
/label=AcGFP1
misc_feature 2801..3595
/label=Kan(R)/Neo(R)
misc_feature 1724..1758
/label=SV40\early\polyA\signals
misc_feature 1821..2276
/label=f1\origin
misc_feature 2338..2366
/label=Bacterial\Promoter\for\Kan(R)
misc_feature 2617..2752
/label=SV40\origin
misc_feature 2450..2679
/label=SV40\early\Promoter/Enhancer
misc_feature 3831..3849
/label=HSV\TK\polyA\signals
misc_feature 4180..4823
/label=pUC\origin
BASE COUNT 1157 a 1341 c 1305 g 1108 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 gctagcatga
601 ggcttcggga gccgctcctg agcggcagcg ccgcgatgcc aggcgcgtcc ctacagcggg
661 cctgccgcct gctcgtggcc gtctgcgctc tgcaccttgg cgtcaccctc gtttactacc
721 tggctggccg cgacctgagc cgcctgcccc aactggtcgg agtctccaca ccgctgcagg
781 gcggctcgaa cagtgccgcc gccatcgggc agtcctccgg ggagctccgg accggagggg
841 ccaaggatcc accggtcatg gtgagcaagg gcgccgagct gttcaccggc atcgtgccca
901 tcctgatcga gctgaatggc gatgtgaatg gccacaagtt cagcgtgagc ggcgagggcg
961 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
1021 ctgtgccctg gcccaccctg gtgaccaccc tgagctacgg cgtgcagtgc ttctcacgct
1081 accccgatca catgaagcag cacgacttct tcaagagcgc catgcctgag ggctacatcc
1141 aggagcgcac catcttcttc gaggatgacg gcaactacaa gtcgcgcgcc gaggtgaagt
1201 tcgagggcga taccctggtg aatcgcatcg agctgaccgg caccgatttc aaggaggatg
1261 gcaacatcct gggcaataag atggagtaca actacaacgc ccacaatgtg tacatcatga
1321 ccgacaaggc caagaatggc atcaaggtga acttcaagat ccgccacaac atcgaggatg
1381 gcagcgtgca gctggccgac cactaccagc agaatacccc catcggcgat ggccctgtgc
1441 tgctgcccga taaccactac ctgtccaccc agagcgccct gtccaaggac cccaacgaga
1501 agcgcgatca catgatctac ttcggcttcg tgaccgccgc cgccatcacc cacggcatgg
1561 atgagctgta caagtgagcg gccgcgactc tagatcataa tcagccatac cacatttgta
1621 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
1681 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
1741 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
1801 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
1861 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
1921 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
1981 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
2041 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
2101 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
2161 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
2221 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
2281 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
2341 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
2401 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
2461 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
2521 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
2581 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
2641 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
2701 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
2761 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
2821 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
2881 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
2941 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
3001 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
3061 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
3121 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
3181 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
3241 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
3301 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
3361 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
3421 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
3481 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
3541 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
3601 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
3661 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
3721 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
3781 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
3841 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
3901 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
3961 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
4021 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
4081 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
4141 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
4201 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
4261 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
4321 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
4381 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
4441 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
4501 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
4561 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
4621 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
4681 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
4741 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
4801 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
4861 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
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