| 出品公司: | Clontech |
|---|---|
| 载体名称: | pProLabel-N |
| 质粒类型: | 哺乳动物表达载体 |
| 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 |
| 载体大小: | 4167 bp |
| 5' 测序引物及序列: | CMV Forward： CGCAAATGGGCGGTAGGCGTG |
| 3' 测序引物及序列: | -- |
| 载体标签: | ProLabel tag（C ter） |
| 载体抗性: | 卡那霉素 |
| 筛选标记: | 新霉素（Neomycin） |
| 克隆菌株: | DH5α 等 |
| 宿主细胞（系）: | 常规细胞系（293、CV-1、CHO等） |
| 备注: | ProLabel tag 是一个分子量约6KD的标签，检测的灵敏度高。 |
| 产品目录号: | 631628 |
| 稳定性: | 稳表达 |
| 组成型/诱导型: | 组成型 |
| 病毒/非病毒: | 非病毒 |
买家导航
pProLabel-N载体质粒基本信息
pProLabel-N质粒图谱载体图谱和pProLabel-N载体序列质粒序列多克隆位点信息
pProLabel-N质粒载体简介
pProLabel-N载体描述
pProLabel-N is a mammalian expression vector designed to express a protein of interest fused to the the n-terminus of a 6 kDa ProLabel tag. The resulting fusion protein can be used in a variety of functional assays and quantitated with our ProLabel Detection Kit II (Cat. No. 631629). The Detection Kit provides all of the components needed to perform enzyme fragment complementation assays (1, 2). In these assays, two inactive enzyme fragments (the ProLabel tag, and a larger Enzyme Acceptor) are combined to form a complete, active enzyme that cleaves the Galacton Star chemiluminescent substrate. The resulting signal can be detected and quantitated with any standard luminometer.
The pProLabel-C vector contains a CMV promoter that drives strong, constitutive expression of the fusion protein, and an SV40 polyadenylation signal that directs processing of the 3’end of the mRNA transcript. The vector also contains a kanamycin/neomycin resistance cassette (Kanr/Neor) that allows G418 selection of stably transfected eukaryotic cells; a bacterial promoter upstream of this cassette allows kanamycin or neomycin selection of transformed bacterial cells. In addition, pProLabel-C contains an SV40 origin of replication for propagation in mammalian cells that express SV40 T-antigen, a pUC origin for propagation in E. coli and an f1 origin for the production of single-stranded DNA.
In order to create a fusion of your protein of interest and the ProLabel tag, your gene of interest must be in the same reading frame as the ProLabel tag sequence, and there can be no intervening stop codons. Prolabel vector constructs can be transfected into mammalian cells using standard transfection methods. ProLabel fusion protein expression levels can be measured quantitatively from mammalian cell lysates using the method described in the ProLabel Detection Kit II Protocol-at-a-Glance (PT3987-2). This highly sensitive and rapid procedure obviates the need for Western analysis. The assay is designed to be used with any ProLabel fusion in a variety of functional assays—from measuring target gene expression in RNAi knockdown studies to measuring protein interactions in coimmunoprecipitation studies.
Propagation in E. coli
Suitable host strains: DH5α, Fusion Blue, 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
参考文献
1. Gorman, C. (1985) In DNA cloning: A practical approach, Vol. II. Ed. D.M. Glover. (IRL Press, Oxford, UK) pp. 143–190.
2. Eglen, R.M. and Singh, R. (2002) Comb. Chem. High Throughput Screen. 6: 381–387.
Note: The attached sequence file has been compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtained by Clontech. This vector has not been completely sequenced.
pProLabel-N质粒序列载体序列
LOCUS pProLabel-N 4167 bp DNA circular 9-NOV-2009
COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT Cat. No. 631628
FEATURES Location/Qualifiers
promoter 1..589
/label=CMV\IE\Promoter
rep_origin 1083..1538
/label=f1\origin
rep_origin 1879..2014
/label=SV40\origin
CDS 2063..2857
/gene="Kanamycin/neomycin resistance gene Neomycin phosphotransferase coding sequences"
/label=Kan(R)/Neo(R)
/note="CDS(Kanamycin/neomycin resistance gene Neomycin phosphotransferase coding sequences)_2"
polyA_signal 3093..3098
/label=HSV\TK\polyA\signal
/note="Herpes simplex virus (HSV) thymidine kinase (TK) polyA signal"
rep_origin 3442..4085
/label=pUC\origin
CDS 666..833
/label=ProLabel\tag
misc_feature 591..665
/label=MCS
polyA_signal 986..1036
/label=SV40\polyA\signal
BASE COUNT 1001 a 1093 c 1079 g 994 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 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
661 gatccagctc caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
721 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
781 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa taggcggccg
841 cgactctaga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
901 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
961 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
1021 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc
1081 gtaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
1141 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
1201 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
1261 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
1321 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
1381 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
1441 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
1501 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa
1561 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
1621 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagtc ctgaggcgga
1681 aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
1741 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
1801 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
1861 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
1921 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
1981 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
2041 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
2101 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
2161 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
2221 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
2281 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
2341 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
2401 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
2461 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
2521 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
2581 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
2641 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
2701 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
2761 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
2821 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
2881 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
2941 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
3001 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
3061 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
3121 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
3181 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
3241 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
3301 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
3361 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
3421 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
3481 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
3541 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
3601 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
3661 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
3721 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
3781 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
3841 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
3901 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
3961 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
4021 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
4081 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
4141 gtggataacc gtattaccgc catgcat
//
pProLabel-N载体图谱质粒图谱pdf版和pProLabel-N载体序列质粒序列等相关资料下载
pProLabel-N质粒载体应用举例
- 上一篇：pProLabel-T
- 下一篇：pMCS-Green-Renilla-Luc
