出品公司: | Clontech |
---|---|
载体名称: | pSEAP2-Control |
质粒类型: | 哺乳动物细胞表达载体;分泌型碱性磷酸酶报告载体 |
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: | 无启动子 |
载体大小: | 5115 bp |
5' 测序引物及序列: | 5'-CTAGCAAAATAGGCTGTCCC-3' |
3' 测序引物及序列: | 5'-CCTCGGCTGCCTCGCGGTTCC-3' |
载体标签: | -- |
载体抗性: | 氨苄青霉素 |
筛选标记: | -- |
克隆菌株: | DH5α 等通用菌株 |
宿主细胞(系): | 常规细胞系(293、CV-1、CHO等) |
备注: |
哺乳动物细胞表达载体pSEAP2-Control没有启动子元件; 利用分泌型碱性磷酸酶定性定量检测启动子及其它顺式转录元件的活性。 |
产品目录号: | 631717 |
稳定性: | 瞬表达 |
组成型/诱导型: | -- |
病毒/非病毒: | 非病毒 |
买家导航
pSEAP2-Control载体质粒基本信息
pSEAP2-Control质粒图谱载体图谱和pSEAP2-Control载体序列质粒序列多克隆位点信息
pSEAP2-Control质粒载体简介
pSEAP2-Control载体描述
pSEAP2-Control is a positive control vector expressing secreted alkaline phosphatase (SEAP) under the control of the SV40 early promoter and the SV40 enhancer. The SEAP coding sequence is followed by the SV40 late polyadenylation signal to ensure proper, efficient processing of the SEAP transcript in eukaryotic cells. A synthetic transcription blocker (TB), composed of adjacent polyadenylation and transcription pause sites, located upstream of the MCS reduces background transcription (1). The vector backbone also contains an f1 origin for single-stran ed DNA production, a pUC origin of replication, and an ampicillin resistance gene for propagation and selection in E. coli. The SEAP2 Vectors incorporate a number of features that improve the sensitivity of SEAP by increasing the efficiency of SEAP expression or that enhance the utility of the vectors. These include: an improved Kozak consensus translation initiation site (2); the removal of the SV40 small-t intron, which can cause cryptic splicing and reduced expression in some genes and/or cell types (3, 4); switching from the early to late polyadenylation signal of SV40, which typically causes a five-fold increase in mRNA levels (5); an expanded multiple cloning site (MCS); compact plasmid size; and removal of extraneous sequences from the 3' untranslated region of the SEAP mRNA.
The integrated set of Great EscAPeTM SEAP2 Vectors has been designed to provide maximal flexibility in studying regulatory sequences from the gene of interest. pSEAP2-Control expresses SEAP in most cell types and provides an important control. The secreted SEAP enzyme is assayed directly from the culture medium and permits time-course studies not possible with assays dependent on cell lysates. Furthermore, the cells can be used for further investigations such as RNA or protein studies. The SEAP2 Vectors can be transfected into mammalian cells by any standard method.
Propagation in E. coli
Suitable host strains: DH5α and other general purpose strains. Single-stranded DNA production requires a host containing an F' episome such as JM109.
Selectable marker: plasmid confers resistance to ampicillin (100 μg/ml) to E. coli hosts.
E. coli replication origin: pUC
Copy number: ~500
Plasmid incompatibility group: pMB1/Col E1
Note: The attached sequence file has been compiled from information in the sequence databases, published literature, and other sources, together with partial sequences obtai ned by Clontech. This vector has not been completely sequenced.
References
1. Eggermont, J. & Proudfoot, N. (1993) EMBO J. 12:2539–2548.
2. Kozak, M. (1987) Nucleic Acids Res. 15:8125–8148.
3. Huang, M. T. F. & Gorman, C. M. (1990) Mol. Cell. Biol. 10:1805–1810.
4. Evans, M. J. & Scarpulla, R. C. (1989) Gene 84:135–142.
5. Carswell, S. & Alwine, J. C. (1989) Mol. Cell. Biol. 9:4248–4258.
6. Levitt, N. et al. (1989) Genes Dev. 3:1019–1025.
7. Enriquez-Harris, P. et al. (1991) EMBO J. 10:1833–1842
pSEAP2-Control质粒序列载体序列
LOCUS pSEAP2-Control 5115 bp DNA circular 5-JAN-2010 COMMENT Created by Clontech Laboratories Inc. http://www.clontech.com COMMENT Cat. No. 631910,631717,631735 FEATURES Location/Qualifiers misc_feature 1..41 /label=MCS\I misc_feature 245..264 /label=MCS\II misc_feature 37..245 /label=SV40\early\Promoter misc_feature 95..230 /label=SV40\origin misc_feature 272..1831 /label=SEAP misc_feature 1942..1947 /label=SV40\late\polyA\signal misc_feature 2056..2301 /label=SV40\enhancer misc_feature 2586..3229 /label=pUC\origin misc_feature complement(3377..4237) /label=Amp(R) misc_feature 4369..4824 /label=f1\origin misc_feature 4955..5108 /label=Transcription\blocker BASE COUNT 1202 a 1392 c 1338 g 1183 t ORIGIN 1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta 61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc 121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc 181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg 241 caaaaagctt cgaatcgcga attcgcccac catgctgctg ctgctgctgc tgctgggcct 301 gaggctacag ctctccctgg gcatcatccc agttgaggag gagaacccgg acttctggaa 361 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc 421 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag 481 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg 541 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg 601 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag 661 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat 721 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca 781 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga 841 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa 901 catggacatt gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc 961 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct 1021 ggtgcaggaa tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct 1081 catgcaggct tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga 1141 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga 1201 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg 1261 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat 1321 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct 1381 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc 1441 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata 1501 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga 1561 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc 1621 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca 1681 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc 1741 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtt actctagagt 1801 cggggcggcc ggccgcttcg agcagacatg ataagataca ttgatgagtt tggacaaacc 1861 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta 1921 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg 1981 tttcaggttc agggggaggt gtgggaggtt ttttaaagca agtaaaacct ctacaaatgt 2041 ggtaaaatcg ataaggatct gaacgatgga gcggagaatg ggcggaactg ggcggagtta 2101 ggggcgggat gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcatgc 2161 tttgcatact tctgcctgct ggggagcctg gggactttcc acacctggtt gctgactaat 2221 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacccta 2281 actgacacac attccacagc ggatccgtcg accgatgccc ttgagagcct tcaacccagt 2341 cagctccttc cggtgggcgc ggggcatgac tatcgtcgcc gcacttatga ctgtcttctt 2401 tatcatgcaa ctcgtaggac aggtgccggc agcgctcttc cgcttcctcg ctcactgact 2461 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac 2521 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa 2581 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg 2641 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa 2701 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc 2761 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac 2821 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac 2881 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg 2941 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt 3001 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa 3061 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct 3121 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga 3181 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg 3241 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct 3301 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt 3361 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc 3421 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg 3481 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag 3541 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt 3601 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag 3661 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt 3721 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca 3781 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg 3841 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat 3901 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta 3961 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca 4021 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct 4081 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat 4141 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa 4201 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt 4261 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa 4321 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta 4381 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca 4441 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct 4501 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc 4561 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat 4621 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc 4681 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc 4741 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta 4801 acaaaatatt aacgcttaca atttgccatt cgccattcag gctgcgcaac tgttgggaag 4861 ggcgatcggt gcgggcctct tcgctattac gccagcccaa gctaccatga taagtaagta 4921 atattaaggt acgggaggta cttggagcgg ccgcaataaa atatctttat tttcattaca 4981 tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca tacgctctcc atcaaaacaa 5041 aacgaaacaa aacaaactag caaaataggc tgtccccagt gcaagtgcag gtgccagaac 5101 atttctctat cgata //
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pSEAP2-Control质粒载体应用举例
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