质粒名称: | pYSEAP |
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出品公司: | 爱迪基因 |
目录编号: | 37326 |
存储实验室: | John Schiller |
载体骨架基本信息 |
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载体名称: | -- |
载体出品公司: | custom |
载体抗性: | 杀稻瘟菌素 75µg/ml |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | 杀稻瘟菌素 |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | SEAP |
物种来源: | H. sapiens (human) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | EF1a |
标签/融合蛋白: | -- |
克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pYSEAP was a gift from John Schiller (Addgene plasmid # 37326 ; http://n2t.net/addgene:37326 ; RRID:Addgene_37326) |
参考文献部分: | Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. Pastrana DV, Buck CB, Pang YY, Thompson CD, Castle PE, FitzGerald PC, Kruger Kjaer S, Lowy DR, Schiller JT. Virology. 2004 Apr 10;321(2):205-16. 10.1016/j.virol.2003.12.027 PubMed 15051381 |
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载体基本信息
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载体序列
LOCUS pYSEAP 5297 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pYSEAP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 5297) AUTHORS Pastrana DV, Buck CB, Pang YY, Thompson CD, Castle PE, FitzGerald PC, Kruger Kjaer S, Lowy DR, Schiller JT TITLE Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18. JOURNAL Virology. 2004 Apr 10;321(2):205-16. PUBMED 15051381 REFERENCE 2 (bases 1 to 5297) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..5297 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 462..1640 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 693..1631 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1588..1608 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1727..3229 /codon_start=1 /product="secreted alkaline phosphatase from human placenta" /label=SEAP /note="The natural enzyme has a C-terminal transmembrane sequence that is replaced by a GPI anchor." /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD" polyA_signal 3287..3408 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3298..3317) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3352..3371 /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind complement(3656..3673) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3827..4415) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3907..3926) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4445..4840) /codon_start=1 /gene="Aspergillus terreus BSD" /product="blasticidin S deaminase" /label=BSD /note="confers resistance to blasticidin" /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI KAIVKDSDGQPTAVGIRELLPSGYVWEG" promoter complement(4859..4906) /label=EM7 promoter /note="synthetic bacterial promoter " promoter complement(4954..5284) /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4968..5103 /label=SV40 ori /note="SV40 origin of replication" primer_bind complement(5022..5041) /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 5269..5289 /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" ORIGIN 1 cgcgttgaca ttgattattg actagttagt tattactagc cgcgtgcaaa gcaccggcgg 61 cggtagatgc ggggtaagta ctgaatttta attcgaccta tcccggtaaa gcgaaagcga 121 cacgcttttt tttcacacat agcgggaccg aacacgttat aagtatcgaa ctcctaaaga 181 agcagtgtag ttttctgcag atgcaaaaaa gatctcatga aggaggaact tgtgcagttt 241 acttaatctg ctttaacaca gctaaaagca gagaaacagt ccggaatctg atggcaaaca 301 cgctaaatgt aagagaagag tgtttgatgc tgcagccagc taaaattcga ggactcagcg 361 cagctctatt ctggtttagc ttgcaaagat ggataaagtt ttaaacagag aggaatcttt 421 gcagctaatg gaccttgtag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt 481 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 541 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 601 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 661 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 721 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccacc tggctgcagt 781 acgtgattct tgatcccgag cttcgggttg gaagtgggtg ggagagttcg aggccttgcg 841 cttaaggagc cccttcgcct cgtgcttgag ttgaggcctg gcctgggcgc tggggccgcc 901 gcgtgcgaat ctggtggcac cttcgcgcct gtctcgctgc tttcgataag tctctagcca 961 tttaaaattt ttgatgacct gctgcgacgc tttttttctg gcaagatagt cttgtaaatg 1021 cgggccaaga tctgcacact ggtatttcgg tttttggggc cgcgggcggc gacggggccc 1081 gtgcgtccca gcgcacatgt tcggcgaggc ggggcctgcg agcgcggcca ccgagaatcg 1141 gacgggggta gtctcaagct ggccggcctg ctctggtgcc tggcctcgcg ccgccgtgta 1201 tcgccccgcc ctgggcggca aggctggccc ggtcggcacc agttgcgtga gcggaaagat 1261 ggccgcttcc cggccctgct gcagggagct caaaatggag gacgcggcgc tcgggagagc 1321 gggcgggtga gtcacccaca caaaggaaaa gggcctttcc gtcctcagcc gtcgcttcat 1381 gtgactccac ggagtaccgg gcgccgtcca ggcacctcga ttagttctcg agcttttgga 1441 gtacgtcgtc tttaggttgg ggggaggggt tttatgcgat ggagtttccc cacactgagt 1501 gggtggagac tgaagttagg ccagcttggc acttgatgta attctccttg gaatttgccc 1561 tttttgagtt tggatcttgg ttcattctca agcctcagac agtggttcaa agtttttttc 1621 ttccatttca ggtgtcgtga ggaattctct agagatccct cgacctcgag atccattgtg 1681 ctggatctgc gatctaagta agcttcgaat cgcgaattcg cccaccatgc tgctgctgct 1741 gctgctgctg ggcctgaggc tacagctctc cctgggcatc atcccagttg aggaggagaa 1801 cccggacttc tggaaccgcg aggcagccga ggccctgggt gccgccaaga agctgcagcc 1861 tgcacagaca gccgccaaga acctcatcat cttcctgggc gatgggatgg gggtgtctac 1921 ggtgacagct gccaggatcc taaaagggca gaagaaggac aaactggggc ctgagatacc 1981 cctggccatg gaccgcttcc catatgtggc tctgtccaag acatacaatg tagacaaaca 2041 tgtgccagac agtggagcca cagccacggc ctacctgtgc ggggtcaagg gcaacttcca 2101 gaccattggc ttgagtgcag ccgcccgctt taaccagtgc aacacgacac gcggcaacga 2161 ggtcatctcc gtgatgaatc gggccaagaa agcagggaag tcagtgggag tggtaaccac 2221 cacacgagtg cagcacgcct cgccagccgg cacctacgcc cacacggtga accgcaactg 2281 gtactcggac gccgacgtgc ctgcctcggc ccgccaggag gggtgccagg acatcgctac 2341 gcagctcatc tccaacatgg acattgacgt gatcctaggt ggaggccgaa agtacatgtt 2401 tcgcatggga accccagacc ctgagtaccc agatgactac agccaaggtg ggaccaggct 2461 ggacgggaag aatctggtgc aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg 2521 gaaccgcact gagctcatgc aggcttccct ggacccgtct gtgacccatc tcatgggtct 2581 ctttgagcct ggagacatga aatacgagat ccaccgagac tccacactgg acccctccct 2641 gatggagatg acagaggctg ccctgcgcct gctgagcagg aacccccgcg gcttcttcct 2701 cttcgtggag ggtggtcgca tcgaccatgg tcatcatgaa agcagggctt accgggcact 2761 gactgagacg atcatgttcg acgacgccat tgagagggcg ggccagctca ccagcgagga 2821 ggacacgctg agcctcgtca ctgccgacca ctcccacgtc ttctccttcg gaggctaccc 2881 cctgcgaggg agctccatct tcgggctggc ccctggcaag gcccgggaca ggaaggccta 2941 cacggtcctc ctatacggaa acggtccagg ctatgtgctc aaggacggcg cccggccgga 3001 tgttaccgag agcgagagcg ggagccccga gtatcggcag cagtcagcag tgcccctgga 3061 cgaagagacc cacgcaggcg aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct 3121 ggttcacggc gtgcaggagc agaccttcat agcgcacgtc atggccttcg ccgcctgcct 3181 ggagccctac accgcctgcg acctggcgcc ccccgccggc accaccgacg ccgcgcaccc 3241 gggttactct agagtcgggg cggccggccg cttcgagcag acatgataag atacattgat 3301 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt 3361 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat 3421 tgcattcatt ttatgtttca ggttcagggg gaggtgtggg aggtttttta aagcaagtaa 3481 aacctctaca aatgtggtaa aatcgataag gatccgtcga ccgatgccct tgagagcctt 3541 caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg cacttatgac 3601 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctcttcc gcttcctcgc 3661 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg 3721 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag 3781 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc 3841 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag 3901 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga 3961 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc 4021 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg 4081 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt 4141 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca 4201 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca 4261 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag 4321 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca 4381 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg 4441 ggtcgccctc ccacacataa ccagagggca gcaattcacg aatcccaact gccgtcggct 4501 gtccatcact gtccttcact atggctttga tcccaggatg cagatcgaga agcacctgtc 4561 ggcaccgtcc gcaggggctc aagatgcccc tgttctcatt tccgatcgcg acgatacaag 4621 tcaggttgcc agctgccgca gcagcagcag tgcccagcac cacgagttct gcacaaggtc 4681 ccccagtaaa atgatataca ttgacaccag tgaagatgcg gccgtcgcta gagagagctg 4741 cgctggcgac gctgtagtct tcagagatgg ggatgctgtt gattgtagcc gttgctcttt 4801 caatgagggt ggattcttct tgagacaaag gcttggccat ggtttagttc ctcaccttgt 4861 cgtattatac tatgccgata tactatgccg atgattaatt gtcaacacgt gctgatcaga 4921 tccgaaaatg gatatacaag ctcccgggag ctttttgcaa aagcctaggc ctccaaaaaa 4981 gcctcctcac tacttctgga atagctcaga ggcagaggcg gcctcggcct ctgcataaat 5041 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg 5101 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgca tgctttgcat 5161 acttctgcct gctggggagc ctggggactt tccacacctg gttgctgact aattgagatg 5221 catgctttgc atacttctgc ctgcctgggg agcctgggga ctttccacac cctaactgac 5281 acacattcca cagaatt //
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