pSEAP2-Basic

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-Basic
质粒类型: 哺乳动物细胞表达载体;无启动子载体;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:
载体大小: 4677 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-Basic没有启动子元件;
利用分泌型碱性磷酸酶定量定性研究外源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-Basic载体图谱



pSEAP2-Basic 多克隆位点

pSEAP2-Basic 载体特征1
pSEAP2-Basic 载体特征2

载体简介

pSEAP2-Basic载体描述

pSEAP2-Basic allows for expression of the reporter gene secreted alkaline phosphatase (SEAP). This vector lacks eukaryotic promoter and enhancer sequences and has an MCS that allows putative promoter DNA fragments to be inserted upstream of the SEAP gene. Enhancers can be cloned into either the MCS or unique downstream sites. 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-stranded 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 increaing 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 EscAPe? SEAP2 Vectors has been designed to provide maximal flexibility in studying regulatory sequences from the gene of interest. pSEAP2-Basic allows for expression of SEAP under control of putative promoters and/or enhancers of interest. 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

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.

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

载体序列

LOCUS       pSEAP2-Basic   4677 bp    DNA   circular             5-JAN-2010
COMMENT     Created by Clontech Laboratories Inc. http://www.clontech.com
COMMENT     Cat. No. 631735,631715
FEATURES             Location/Qualifiers
     misc_feature    1..72
                     /label=MCS
     misc_feature    80..1639
                     /label=SEAP
     misc_feature    1750..1755
                     /label=SV40\late\polyA\signal
     misc_feature    2148..2791
                     /label=pUC\origin
     misc_feature    complement(2939..3799)
                     /label=Amp(R)
     misc_feature    3931..4386
                     /label=f1\origin
     misc_feature    4517..4670
                     /label=Transcription\blocker
BASE COUNT     1117 a      1270 c      1216 g      1074 t 
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctaa gtaagcttcg 
       61 aatcgcgaat tcgcccacca tgctgctgct gctgctgctg ctgggcctga ggctacagct 
      121 ctccctgggc atcatcccag ttgaggagga gaacccggac ttctggaacc gcgaggcagc 
      181 cgaggccctg ggtgccgcca agaagctgca gcctgcacag acagccgcca agaacctcat 
      241 catcttcctg ggcgatggga tgggggtgtc tacggtgaca gctgccagga tcctaaaagg 
      301 gcagaagaag gacaaactgg ggcctgagat acccctggcc atggaccgct tcccatatgt 
      361 ggctctgtcc aagacataca atgtagacaa acatgtgcca gacagtggag ccacagccac 
      421 ggcctacctg tgcggggtca agggcaactt ccagaccatt ggcttgagtg cagccgcccg 
      481 ctttaaccag tgcaacacga cacgcggcaa cgaggtcatc tccgtgatga atcgggccaa 
      541 gaaagcaggg aagtcagtgg gagtggtaac caccacacga gtgcagcacg cctcgccagc 
      601 cggcacctac gcccacacgg tgaaccgcaa ctggtactcg gacgccgacg tgcctgcctc 
      661 ggcccgccag gaggggtgcc aggacatcgc tacgcagctc atctccaaca tggacattga 
      721 cgtgatccta ggtggaggcc gaaagtacat gtttcgcatg ggaaccccag accctgagta 
      781 cccagatgac tacagccaag gtgggaccag gctggacggg aagaatctgg tgcaggaatg 
      841 gctggcgaag cgccagggtg cccggtatgt gtggaaccgc actgagctca tgcaggcttc 
      901 cctggacccg tctgtgaccc atctcatggg tctctttgag cctggagaca tgaaatacga 
      961 gatccaccga gactccacac tggacccctc cctgatggag atgacagagg ctgccctgcg 
     1021 cctgctgagc aggaaccccc gcggcttctt cctcttcgtg gagggtggtc gcatcgacca 
     1081 tggtcatcat gaaagcaggg cttaccgggc actgactgag acgatcatgt tcgacgacgc 
     1141 cattgagagg gcgggccagc tcaccagcga ggaggacacg ctgagcctcg tcactgccga 
     1201 ccactcccac gtcttctcct tcggaggcta ccccctgcga gggagctcca tcttcgggct 
     1261 ggcccctggc aaggcccggg acaggaaggc ctacacggtc ctcctatacg gaaacggtcc 
     1321 aggctatgtg ctcaaggacg gcgcccggcc ggatgttacc gagagcgaga gcgggagccc 
     1381 cgagtatcgg cagcagtcag cagtgcccct ggacgaagag acccacgcag gcgaggacgt 
     1441 ggcggtgttc gcgcgcggcc cgcaggcgca cctggttcac ggcgtgcagg agcagacctt 
     1501 catagcgcac gtcatggcct tcgccgcctg cctggagccc tacaccgcct gcgacctggc 
     1561 gccccccgcc ggcaccaccg acgccgcgca cccgggttac tctagagtcg gggcggccgg 
     1621 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg 
     1681 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt 
     1741 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag 
     1801 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat 
     1861 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc 
     1921 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg 
     1981 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc 
     2041 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag 
     2101 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa 
     2161 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc 
     2221 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc 
     2281 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg 
     2341 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt 
     2401 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc 
     2461 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc 
     2521 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag 
     2581 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg 
     2641 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa 
     2701 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag 
     2761 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact 
     2821 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa 
     2881 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt 
     2941 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag 
     3001 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca 
     3061 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc 
     3121 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt 
     3181 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg 
     3241 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca 
     3301 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg 
     3361 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca 
     3421 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg 
     3481 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct 
     3541 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca 
     3601 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca 
     3661 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg 
     3721 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac 
     3781 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt 
     3841 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc 
     3901 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg 
     3961 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc 
     4021 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa 
     4081 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac 
     4141 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt 
     4201 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca 
     4261 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt 
     4321 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta 
     4381 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct 
     4441 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg 
     4501 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt 
     4561 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact 
     4621 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgata 
//

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