| 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
载体基本信息
载体图谱质粒图谱和多克隆位点信息
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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