质粒名称: | pT7-NSP5SA11 |
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出品公司: | Addgene |
目录编号: | 89172 |
存储实验室: | Takeshi Kobayashi |
载体骨架基本信息 |
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载体名称: | p3E5 |
载体出品公司: | -- |
空载体大小: | 3074 bp |
完整质粒大小: | 3741 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | -- |
载体抗性: | 氨苄霉素 |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | Rotavirus SA11 NSP5 |
物种来源: | Rotavirus SA11 |
插入大小: | 667 bp |
基因突变: | -- |
GenBank编号 | LC178574 |
启动子: | T7 promotor |
标签/融合蛋白: | -- |
克隆方法: | Gibson克隆 |
5’克隆位点: | -- |
3’克隆位点: | -- |
5’测序引物 | T7 forward CTGTGGATAACCGTATTACCG |
3’测序引物 | T7 terminal GCTAGTTATTGCTCAGCGG |
文献引用方法 |
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材料和方法部分: | pT7-NSP5SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89172 ; http://n2t.net/addgene:89172 ; RRID:Addgene_89172) |
参考文献部分: | Entirely plasmid-based reverse genetics system for rotaviruses. Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N, Onishi M, Matsuura Y, Taniguchi K, Kobayashi T. Proc Natl Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 10.1073/pnas.1618424114. 10.1073/pnas.1618424114 PubMed 28137864 |
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pT7NSP5SA11载体基本信息
pT7NSP5SA11载体图谱质粒图谱和多克隆位点信息
pT7NSP5SA11载体简介
pT7NSP5SA11载体序列
LOCUS pT7-NSP5SA11 3740 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express Rotavirus SA11 NSP5 in mammalian cells. ACCESSION . VERSION . KEYWORDS pT7-NSP5SA11 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 3740) AUTHORS Kanai Y, Komoto S, Kawagishi T, Nouda R, Nagasawa N, Onishi M, Matsuura Y, Taniguchi K, Kobayashi T TITLE Entirely plasmid-based reverse genetics system for rotaviruses. JOURNAL Proc Natl Acad Sci U S A. 2017 Jan 30. pii: 201618424. doi: 10.1073/pnas.1618424114. PUBMED 28137864 REFERENCE 2 (bases 1 to 3740) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..3740 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 33..44 /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" promoter complement(90..108) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(192..209) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(363..951) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(443..462) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(1122..1982) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 1745..1764 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(1983..2087) /gene="bla" /label=AmpR promoter primer_bind 2155..2173 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(2211..2233) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 2333..2352 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 2562..2581 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 2871..2889 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 2871..2888 /label=SP6 /note="SP6 promoter, forward primer" terminator 2957..3004 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 3000..3018 /label=T7 Term /note="T7 terminator, reverse primer" ORIGIN 1 aagacgattc attcttatat atagttgaag gaatagaagg aagactcgtc acgtcaatac 61 tgagagacat cactgtagcg ctttaaaagc ctatagtgag tcgtattaac cgggatcgat 121 atccccctgc attaatgaat cggccaatgc gcggggagag gcggtttgcg tattgggcgc 181 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 241 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 301 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 541 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 781 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 1021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 1081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 1141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 1201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 1261 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 1321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 1381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 1441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 1501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 1561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 1621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 1681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 1741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 1801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 1861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 1921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 1981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 2041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 2101 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 2161 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 2221 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc 2281 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca 2341 gagcagattg tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattaa 2401 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcaag 2461 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa 2521 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag 2581 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg 2641 atctggctag cgatgaccct gctgattggt tcgctgacca tttccgggtg cgggacggcg 2701 ttaccagaaa ctcagaaggt tcgtccaacc aaaccgactc tgacggcagt ttacgagaga 2761 gatgataggg tctgcttcag taagccagat gctacacaat taggcttgta catattgtcg 2821 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga 2881 cactatagaa tacaagcttg catgcctgca ggtcgactct agaggatctc gatccggata 2941 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg 3001 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag 3061 ccggatcctt ttttttgagc tctcccttag ccatccgagt ggacgacgtc ctccttcgga 3121 tgcccaggtc ggaccgcgag gaggtggaga tgccatgccg acccggtcac aaaacgggag 3181 tggggagctc cctagtgtgt tcccaggtca gacttacaaa tcttcaatca attgcattgc 3241 gacttgtttc attctcattc ttaatgcgaa gtatttcttc ttatatttac agttcttaca 3301 tttaccatca tcactatctg aatcatccag tacataatca tctgaatcag actctgcttc 3361 aattcttggg tagtgcttcc tacttttatg gttttgtttt gatttctcct ttttagtatc 3421 cgtagatatt gatattgatg agtccaaatt agcttttact tttaagcctt tatttaagga 3481 gaaatccact tgatcgcatc cgacattact tgaaggtcgt gattgtgctg atgaatccat 3541 agacacgcca gcgtctgcgt ttgtcttaac tgcattcgat ctaatcgaaa aactggtgag 3601 tggatcgttt gaagcagaat cagatggtcc aatatcctct ggagacttcg acagcatgta 3661 tttattgaat gcttctgcat ctggtgaaat gtactgttca ctcctaccaa tagattttcc 3721 agaaagagtt gacgttgttg //
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