| 质粒名称: | pT7-NSP3SA11 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89170 |
| 存储实验室: | Takeshi Kobayashi |
|
载体骨架基本信息 |
|
| 载体名称: | p3E5 |
| 载体出品公司: | -- |
| 空载体大小: | 3074 bp |
| 完整质粒大小: | 4179 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 |
|
| 插入基因名称: | Rotavirus SA11 NSP3 |
| 物种来源: | Rotavirus SA11 |
| 插入大小: | 1059 bp |
| 基因突变: | |
| GenBank编号 | LC178572 |
| 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 |
|
| 材料和方法部分: | pT7-NSP3SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89170 ; http://n2t.net/addgene:89170 ; RRID:Addgene_89170) |
| 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
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
载体序列
LOCUS pT7-NSP3SA11 4179 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Rotavirus SA11 NSP3 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pT7-NSP3SA11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4179)
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 4179)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4179
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(30..47)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(201..789)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(281..300)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(960..1820)
/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 1583..1602
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1821..1925)
/gene="bla"
/label=AmpR promoter
primer_bind 1993..2011
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2049..2071)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2171..2190
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2400..2419
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2709..2727
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2709..2726
/label=SP6
/note="SP6 promoter, forward primer"
terminator 2795..2842
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2838..2856
/label=T7 Term
/note="T7 terminator, reverse primer"
promoter complement(4106..4124)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
61 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
361 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
601 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
1021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
1081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
1141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
1201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
1261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
1321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
1381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
1441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
1501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
1561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
1621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
1681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
1741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
1801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
1861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
1921 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
1981 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
2041 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
2101 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
2161 cttaactatg cggcatcaga gcagattgta ctgagagtgc accattcgac gctctccctt
2221 atgcgactcc tgcattaaga agcagcccag tagtaggttg aggccgttga gcaccgccgc
2281 cgcaaggaat ggtgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
2341 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
2401 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
2461 atgcgtccgg cgtagaggat ctggctagcg atgaccctgc tgattggttc gctgaccatt
2521 tccgggtgcg ggacggcgtt accagaaact cagaaggttc gtccaaccaa accgactctg
2581 acggcagttt acgagagaga tgatagggtc tgcttcagta agccagatgc tacacaatta
2641 ggcttgtaca tattgtcgtt agaacgcggc tacaattaat acataacctt atgtatcata
2701 cacatacgat ttaggtgaca ctatagaata caagcttgca tgcctgcagg tcgactctag
2761 aggatctcga tccggatata gttcctcctt tcagcaaaaa acccctcaag acccgtttag
2821 aggccccaag gggttatgct agttattgct cagcggtggc agcagccaac tcagcttcct
2881 ttcgggcttt gttagcagcc ggatcctttt ttttgagctc tcccttagcc atccgagtgg
2941 acgacgtcct ccttcggatg cccaggtcgg accgcgagga ggtggagatg ccatgccgac
3001 ccggccacat aacgccccta tagccattta ggtttttgac agtgttagct tttaactatt
3061 gtgctcatag agggttatgc atagatgaca acatactaac actgtatttg attgttcaac
3121 atgtatcaaa atggttattc gcaaccaaat gaatattgat aattacatct ctgtattaat
3181 cctttgaaca taaggaataa tttgtcataa tcagatatca aattacgtat tactgattct
3241 aagtcatcaa aagcgtgcaa tggatttatc gcatcaattg agttaatttg ctgttcaatg
3301 tctgccttta tttcagggtc taattccatt gatcttaaat accattcaat cgaagaagta
3361 agggatccaa ttttattttg caaatcacgt tctagtttat tattgtacac ttgaagctca
3421 gctatatgtg cttgctgttg agagatgaca ttttgaagag aatgcatatt ttcattcatt
3481 tttcttgctt tcaacaccca attattatat ttttcattta ccctggattt caatgattca
3541 aacctttgct ccaattgctc atagcgcgat ttccagtcaa tggtatccac ttccattttt
3601 tcatccacaa atgaatcatc cacttcaact tcaccacgtt ccaatttatc acgcatcaat
3661 tttgtgcatt taataataga tgatgatttt ccaggtattc tttttacact gaagcatgcg
3721 tttaaaactc tcattttttg atcaattcct tttgatgata acatcattct tagtttgttt
3781 acatcctcat ctaatttagc tgctctagaa gtatcagcaa gccagtttct atttcttata
3841 gcagatccaa atttattatt caaagcttga tcaatagtta ttgctttacc aatcagatta
3901 tttttaacac cagaatcgtc catcacaaaa tcaaatttat tctttactct agaatatata
3961 tcctgataat catattctat tcccatattc tcaagagctg aggttgcagc tacaactgca
4021 gcttcaaaag aagagttaat aattgagacg gccatctgtt gcgtagactc catcttgagc
4081 atcaaccact gaaaagcatt aaatgcctat agtgagtcgt attaacccgg gatcgatatc
4141 cccctgcatt aatgaatcgg ccaacgcgcg gggagaggc
//
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