质粒名称: | pT7-NSP1SA11 |
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出品公司: | Addgene |
目录编号: | 89168 |
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: | 4684 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 NSP1 |
物种来源: | Rotavirus SA11 |
插入大小: | 1610 bp |
基因突变: | -- |
GenBank编号 | LC178570 |
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-NSP1SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89168 ; http://n2t.net/addgene:89168 ; RRID:Addgene_89168) |
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 |
买家导航
pT7NSP1SA11载体基本信息
pT7NSP1SA11载体图谱质粒图谱和多克隆位点信息
pT7NSP1SA11载体简介
pT7NSP1SA11载体序列
LOCUS pT7-NSP1SA11 4684 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express Rotavirus SA11 NSP1 in mammalian cells. ACCESSION . VERSION . KEYWORDS pT7-NSP1SA11 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4684) 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 4684) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..4684 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 18..36 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(74..96) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind 196..215 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 425..444 /label=pBRrevBam /note="pBR322 vectors, tet region, downstream of BamHI, reverse primer" promoter 734..752 /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind 734..751 /label=SP6 /note="SP6 promoter, forward primer" terminator 820..867 /label=T7 terminator /note="transcription terminator for bacteriophage T7 RNA polymerase" primer_bind 863..881 /label=T7 Term /note="T7 terminator, reverse primer" promoter complement(2636..2654) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(2739..2756) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(2910..3498) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(2990..3009) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(3669..4529) /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 4292..4311 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(4530..4634) /gene="bla" /label=AmpR promoter ORIGIN 1 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg 61 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag 121 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg 181 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct 241 cccttatgcg actcctgcat taagaagcag cccagtagta ggttgaggcc gttgagcacc 301 gccgccgcaa ggaatggtgc aaggagatgg cgcccaacag tcccccggcc acggggcctg 361 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc 421 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg 481 ccacgatgcg tccggcgtag aggatctggc tagcgatgac cctgctgatt ggttcgctga 541 ccatttccgg gtgcgggacg gcgttaccag aaactcagaa ggttcgtcca accaaaccga 601 ctctgacggc agtttacgag agagatgata gggtctgctt cagtaagcca gatgctacac 661 aattaggctt gtacatattg tcgttagaac gcggctacaa ttaatacata accttatgta 721 tcatacacat acgatttagg tgacactata gaatacaagc ttgcatgcct gcaggtcgac 781 tctagaggat ctcgatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg 841 tttagaggcc ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc 901 ttcctttcgg gctttgttag cagccggatc cttttttttg agctctccct tagccatccg 961 agtggacgac gtcctccttc ggatgcccag gtcggaccgc gaggaggtgg agatgccatg 1021 ccgacccggt tcacagtatt ttgccagcta ggcgctactc tagtctggac ggtcttgtga 1081 tggctaaata ctgtataatt agatagtgac ataatttcac tcattgtcat cttctgagtt 1141 ggagatcagt agctcaaatt cttcagttaa agttccagaa ttcttcattt caatcagttc 1201 ctccgttgat tttattgaca ttggcaattt atccaattca tcaacattat accatttact 1261 gctaattgat ttaactatac tactgaattc atttaaatta agtaccattg gtatgctatc 1321 tacattcaaa gtgcgcttaa tttcattata caaactaata ttcaatccaa gaatcaacat 1381 gatgacattc gtaccttcaa ctgacactgg cgacatgtga ttaaacactt cttcaattgt 1441 ttgatccatg tgattagtaa acaaactcca aattaatctt ttaattaaag gatatatgtt 1501 ttcaacagaa ctacaatgtc caacgttaac attactttta taacaatcct taataaattc 1561 catcaataaa ttatacatat cacgaatccg ccaatcaatt ttactaagaa aatattttat 1621 attacaccat ttacagtact ggatatcgga agcattgcct gggtgattag atgccacgta 1681 gttaggctga caatgatgtc ccaactcaaa tattgtcatt ttcaaccttc caaaattatt 1741 atcatgagca tagaaccgag tatacttact cgtcatcatt gcagcaatta atttattctt 1801 atcatttttt aaatcagaat aaagctccaa ttcatctctc caattaaatt cagttacatt 1861 tctgtatatt gatcttcctg gcgtttttgt ataagtatta aaaatcacag tttctttagt 1921 aaaatctgaa aagtaaattt gctttagttt tgatgatggc agtcttgata attggaagta 1981 tttctttgcg taatggtttc tcaactctat tggtagattg gacattcttg taaagttaat 2041 atgatcaagc aataatttat caaattcatt tataaagtta acaattctat atggagcata 2101 tatatcactg actgatttct catataaccc aaaaacataa taaagattat tttcatcaaa 2161 cttaaatgca gcagcattta atgtaattgg gagcagcaaa tgattatacc attcaatttt 2221 atattgattc ctacattctc tttgctttat cattctttca aattttctaa ttactttatc 2281 attaattgga aatattatat tgtacaagtc tatgatacac tgtaatttgt ctttggtaat 2341 tggatgtttc acagttctaa gcttaagcaa atgaggttca ttgtcaagga agcatcttct 2401 gttctgaaca caccatttgc aaacatgcgg caatgagcaa ccataacaat gggttaaatc 2461 agctatttga caacattcta aacaccaccc cttaattttc gcatcaggaa ctggcatcca 2521 aatagaattt gcaccaatgt tgcataatct ccgatttaaa gcagttaatc tacgataatg 2581 aaagcatgca tctttaaaag tagccatggc taacacaaga cttttcaaaa aaaagcctat 2641 agtgagtcgt attaacccgg gatcgatatc cccctgcatt aatgaatcgg ccaacgcgcg 2701 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc 2761 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc 2821 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg 2881 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat 2941 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag 3001 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga 3061 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg 3121 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt 3181 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac 3241 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc 3301 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt 3361 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc 3421 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc 3481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg 3541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag 3601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg 3661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt 3721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca 3781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca 3841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc 3901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt 3961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg 4021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc 4081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg 4141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga 4201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga 4261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta 4321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg 4381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact 4441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata 4501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt 4561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa 4621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt 4681 atca //
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