| 质粒名称: | pT7-VP4SA11 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89165 |
| 存储实验室: | Takeshi Kobayashi |
|
载体骨架基本信息 |
|
| 载体名称: | p3E5 |
| 空载体大小: | 3074 bp |
| 完整质粒大小: | 5436 bp |
| 载体类型: | 哺乳细胞表达 |
| 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 VP4 |
| 物种来源: | Rotavirus SA11 |
| 插入大小: | 2362 bp |
| GenBank编号 | LC178567 |
| 启动子: | T7 promotor |
| 克隆方法: | Gibson克隆 |
| 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-VP4SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89165 ; http://n2t.net/addgene:89165 ; RRID:Addgene_89165) |
| 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
pT7VP4SA11载体基本信息
pT7VP4SA11载体图谱质粒图谱和多克隆位点信息
pT7VP4SA11载体简介
pT7VP4SA11载体序列
LOCUS pT7-VP4SA11 5436 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Rotavirus SA11 VP4 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pT7-VP4SA11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5436)
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 5436)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5436
/organism="synthetic DNA construct"
/mol_type="other DNA"
rep_origin complement(48..636)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(128..147)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(807..1667)
/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 1430..1449
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1668..1772)
/gene="bla"
/label=AmpR promoter
primer_bind 1840..1858
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(1896..1918)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2018..2037
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2247..2266
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2556..2574
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2556..2573
/label=SP6
/note="SP6 promoter, forward primer"
terminator 2642..2689
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2685..2703
/label=T7 Term
/note="T7 terminator, reverse primer"
promoter complement(5210..5228)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(5313..5330)
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
61 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
121 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
181 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
241 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
301 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
361 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
421 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
481 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
541 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
601 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
661 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
721 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
781 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
841 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
901 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
961 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
1021 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
1081 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
1141 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
1201 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
1261 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
1321 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
1381 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
1441 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
1501 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
1561 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
1621 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
1681 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
1741 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
1801 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
1861 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
1921 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
1981 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
2041 agagtgcacc attcgacgct ctcccttatg cgactcctgc attaagaagc agcccagtag
2101 taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcaaggagat ggcgcccaac
2161 agtcccccgg ccacggggcc tgccaccata cccacgccga aacaagcgct catgagcccg
2221 aagtggcgag cccgatcttc cccatcggtg atgtcggcga tataggcgcc agcaaccgca
2281 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatctg gctagcgatg
2341 accctgctga ttggttcgct gaccatttcc gggtgcggga cggcgttacc agaaactcag
2401 aaggttcgtc caaccaaacc gactctgacg gcagtttacg agagagatga tagggtctgc
2461 ttcagtaagc cagatgctac acaattaggc ttgtacatat tgtcgttaga acgcggctac
2521 aattaataca taaccttatg tatcatacac atacgattta ggtgacacta tagaatacaa
2581 gcttgcatgc ctgcaggtcg actctagagg atctcgatcc ggatatagtt cctcctttca
2641 gcaaaaaacc cctcaagacc cgtttagagg ccccaagggg ttatgctagt tattgctcag
2701 cggtggcagc agccaactca gcttcctttc gggctttgtt agcagccgga tccttttttt
2761 tgagctctcc cttagccatc cgagtggacg acgtcctcct tcggatgccc aggtcggacc
2821 gcgaggaggt ggagatgcca tgccgacccg gtcacatcct ctagaaatta ctcacaacct
2881 gcattgcata atcagttgtt caattctatt tctaattata ggattgtcct gattaatgaa
2941 ttcacgtaaa actcttgggt cagatcttaa aagattaaat gcttgttgct tagtaatgcc
3001 gtaattatcg ttcaaatttt taagtgtttt aaaatcaatt atagcggata ttactggaga
3061 atctgtaact aagtcagcga atttttgtac atcaaatggt atttcctcaa atgtatccac
3121 tttataagca aaaaatttcc catcaattcc agcttcaaac acatcatcgt tgtttataac
3181 gcggtaagcc ctatttggta tgaatttttc cgatgcttca gtaacaatgt caggtattgt
3241 gtttggtgat atttgagtcg atttatctat cttagtcttc aaaacagccg ctgatatatc
3301 atcaaaattc ataccctcag tttgtgttgc catttccttt agtctcaatc ttctactgat
3361 agttgacgtt tgtgtcgaaa ctgaacttac tgatgacgat atatcagtta tttgtgttga
3421 tacatccgtc catgctgatg ctgaagatcc aatcgaacgt atagatgaac ctcttgatat
3481 tgatgatgct gcgtctgata aagaatctgt cagtgttgaa actgaattcg ctaatcctga
3541 ctttttgaat tttttcataa cattagtagc cattgatttt gcagcatcaa tagtactttt
3601 aatgccagaa aacattgaga acatatctaa tggtagaagc gctaaatcga ttaactgcga
3661 cattgcaatt tcttgagaca aagcgttaaa ctcttctctt agttctccta actgcctttc
3721 taaatcttgt cgtacagtaa ctgagtttgc tataggagtc tgatagtcat catttgatgg
3781 cactaatgat ataagtgaaa atcgtccagc aatttcataa tattctttac cattatttgg
3841 atctgcagca ggcagaccat acaatctatc taatctagtt ctagtcagtt taaagtgtgg
3901 ttcttcgaca gctagtctaa atctaaatct taatgaattt aatgatacga aatctgtaaa
3961 ttgagtagat agtgttacac cagctgaatg taaagaaact gctcccccag ttaaaacagg
4021 ccattgtcca actggtagac taaaattata gctgcctcca gtacacataa ctgaattcaa
4081 gtttgctgct aacgatcgga catacaccac gttacgaaat gcttgtgaat catcccagta
4141 atcgatatag acgtatgaat tctctttaat tacttcaaat ttagatacaa caaaatcagt
4201 tggtaaatat cctccattaa aactgaagtc attaacgcca ttcactgaac aagtagtatg
4261 tgcggtaact tcttcaccat cacgagtata tgtgtattga taattcgctg gcttaaatga
4321 tatttctgac cacttatatc ccagccctcc tgattttata attgtatttg caaatttaaa
4381 tctaatagtt atatctctat tatattgcat ttctttccat aatgatgtct tggatatcac
4441 aatatcttca ttagcttgag ctctatagtg tattacatct ctagcagtca acgataatgg
4501 tacaacattt ctagtatttt gtattggtgg taatccatta ttaatgtatt ccgtacattt
4561 agactcttca gatctaggaa ttatataaaa gtcacaaaaa gcagtcatgt taacagaatc
4621 ataattcgtt gttgaataat gtgctgtcct agcgttaggt gtctgtcctt catatgtata
4681 taatttttca ttatgcttca taactgcata taatttcgga ctggatagta atggtccata
4741 ttgtcctata cttccatttg cagttgtttt tacgacatca ataaatttcc actggtcttg
4801 tgaagtattg gataccgtta attgttcttg aataccaaat atagtgtaag ttctattttc
4861 agactgaaca tttggctcga ttaaaattgt ggctaaccat ctatctgtat tatttgtacc
4921 ttcaacaatt acgccaggtg tcgttggagc cagtaacatc caataatcga ctggtggatt
4981 gaatgtcgtt ggttgataag gcccatccag caatggtcca actgtcgtag aatcattaat
5041 ttctccaggt ccccagttaa ctggagcata acctgtttgc gcaaatggtc caggattaat
5101 tgtgacattt tgtgatttag ttgatccaat ctcttgtatc tcatcggata aatctactgt
5161 ataagaattc gtgagcaatt gtctataaat gagcgaagcc attttatagc ctatagtgag
5221 tcgtattaac ccgggatcga tatccccctg cattaatgaa tcggccaacg cgcggggaga
5281 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
5341 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
5401 tcaggggata acgcaggaaa gaacatgtga gcaaaa
//
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