pSEAP2-Control

价格:2500元

联系方式:I47-825O-882O

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载体基本信息

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

载体质粒图谱和多克隆位点信息

pSEAP2-Control载体图谱



pSEAP2-Control 多克隆位点

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

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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