| 质粒名称: | pT7-VP2SA11 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89163 |
| 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: | 5767 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 VP2 |
| 物种来源: | -- |
| 插入大小: | 2693 bp |
| 基因突变: | -- |
| GenBank编号 | LC178565 |
| 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-VP2SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89163 ; http://n2t.net/addgene:89163 ; RRID:Addgene_89163) |
| 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-VP2SA11 5766 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Rotavirus SA11 VP2 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pT7-VP2SA11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5766)
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 5766)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..5766
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(19..36)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(190..778)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(270..289)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(949..1809)
/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 1572..1591
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1810..1914)
/gene="bla"
/label=AmpR promoter
primer_bind 1982..2000
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2038..2060)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2160..2179
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2389..2408
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2698..2716
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2698..2715
/label=SP6
/note="SP6 promoter, forward primer"
terminator 2784..2831
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2827..2845
/label=T7 Term
/note="T7 terminator, reverse primer"
promoter complement(5683..5701)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
61 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
121 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
181 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
241 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
301 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
361 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
421 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
481 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
541 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
601 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
661 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
721 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
781 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
841 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
901 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
961 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
1021 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
1081 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
1141 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
1201 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
1261 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
1321 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
1381 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
1441 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
1501 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
1561 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
1621 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
1681 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
1741 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
1801 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
1861 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
1921 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
1981 aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct
2041 ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag
2101 acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc ttaactatgc
2161 ggcatcagag cagattgtac tgagagtgca ccattcgacg ctctccctta tgcgactcct
2221 gcattaagaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
2281 gtgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
2341 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
2401 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
2461 gtagaggatc tggctagcga tgaccctgct gattggttcg ctgaccattt ccgggtgcgg
2521 gacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga cggcagttta
2581 cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag gcttgtacat
2641 attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac acatacgatt
2701 taggtgacac tatagaatac aagcttgcat gcctgcaggt cgactctaga ggatctcgat
2761 ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga ggccccaagg
2821 ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt tcgggctttg
2881 ttagcagccg gatccttttt tttgagctct cccttagcca tccgagtgga cgacgtcctc
2941 cttcggatgc ccaggtcgga ccgcgaggag gtggagatgc catgccgacc cggtcatatc
3001 tccacaatgg ggttggcgtt tacagttcgt tcatgatgcg catattatca aaagcaacag
3061 cattaattgg ttcaacagta tcagctgaaa cgaacgcaag caaatctgaa tatacggtaa
3121 atgttaggtt agacgttaac atatgcattg acgctctaaa atcaaattgt tgtggaactt
3181 gtttatacac ttttgtagta gatgtaggaa tccaatcata attagccacc aaataaaagt
3241 cattagaatc tgaatttatc ttatatagga tgggttttaa cgtgtcgacc tttctaagtt
3301 tgacaatctg tgcaaaaacg gttgcatcta gtttagctat taacgaaatc actgatgaat
3361 ccgttataaa tggtagcgct ccaactaaag ctactggttg attattaagt agcatgttag
3421 taatttgagc ataatctcct gatctcatca attcttcaag atttatttgt tgaaatccgt
3481 ccaagtttct ggcaatattg acgtaaccat acatctcatc tcgttctaac tgcatatctc
3541 ggtatgctat tataactcct tgtgcaattt tatctgatgc acgttcaatc tgttccatat
3601 tcattgctat caaattaaaa atatctaatc ttcttatttc aactggtaat agtcttaatc
3661 tatctctcaa tctatacatt tggtcatctg gtactctcgc aacatcaaat atctgtaatc
3721 ttttcagaaa gtcctcaact attgatttca ttttcttttg atataaattt aatctatttg
3781 ccgcagttat aattgcaact gcgtcattaa ttctttcatt ataatttgaa tgaaaattga
3841 cattcacatt atagtaatgg aacaatgttt gcggactcgg tataaccgta gcatttccaa
3901 ttagcataca taatgatgtt actgatgtta attgcaattt ttcagttgtt aatgtttgaa
3961 catgctgcat attcattgtt atgcatgcca ttaatgtctc ataattgtat gataacaatc
4021 ttgttaaatc gacaagctga ccaagtctgt tagaaagcag caaaattcct ctctgtatag
4081 atcttttata atcaactggc attgtgggaa actgttgtct agataattgc atcagagctt
4141 ccattaattg gttgactaca tgaccatttc ttatattatc attcaagact tgatttaaca
4201 ctccatcaat cactacttgt ctaaactgat tataattaac aaaatgtaac caattagcta
4261 cctgaaaatt ttgaatctgt tgctctgcaa tttgaaaggg agtctgtgga tcaccattcc
4321 tataatgcat tctttgcatt ccgaatgccg gataaacaat ggtatttatt atggctagtt
4381 gacatgctac taatgattct cttataaaca tatcatttgg aatcacagtg agtaaccaca
4441 tgcctgaaat aagtgacatg taatttgtcg ttacgaaatc taatgacatg gttctctgac
4501 tcaacatagc agcaatcaaa gttttaaaac aatcattagc ggcttgtgag tttatacctg
4561 ttaaaaactg agtctctgat tgtattgtca aagcttccaa ttgcaaatct tgtgacattt
4621 tctggatttg tgcctcagta gatactaatt cttttaggtc tggcaccaca gatcttgcta
4681 aaatataatt agatgtagtt atagtatccc agagtgactc aaaattatca tgtaaattta
4741 acctatcttg tagcaagttt ggtctgatat atctagcagt agacggtaaa ttcctgtcca
4801 tatttaatat atagttgaca tcatttctta ttctctctgg tatgtaattg aaaatgatat
4861 cattatttaa tggttctacc aactgatgtt gtaagaagta ttcgttaaac gcatggtcaa
4921 ttggatgcaa tatagatgga taattgacta catttcgatc agcttgaact cgttgtctca
4981 tctcagctat gaatcttctt actgcacctt cggtttcttc atcttgaaaa atagcatcgc
5041 atattgcggc tgtttcagaa tcaactactt tgccagcatc ccttgaattt ttattctcta
5101 cagccatatc tttaagtaat agataatccg gcatttccat taatacttga tcatataaat
5161 ttaaaaaata ctctctaaca tcataatctc catcaggaag agtatctcgt ttcaatttcc
5221 aataccatct attacgaagc tctctttctc cattagctct ataaataggc aattgtttcg
5281 gttcaaatat tcgaaacagt ttagtttgtt tctttgcttg ttctggtttt atgtcttcta
5341 attttttgag tattgactct tttggttcaa atgtagggat agttttttgt aatatttcat
5401 actgaacttc tttttgatgt tcctcttttg ttttaagtac ttctaacaac tgtttcgatt
5461 cttctttatt agattttttt acctcatcag atatcttaac ttcttcttga ttatctgtaa
5521 ttatctcttc tttcttagat aatacttttt ctgataattg tgatttagga ctgtcattat
5581 taatgttctt gctatcttct ttttcttgca ttcgttcatc ttgttttaga ttcgtctcac
5641 gacgcgctcc acgttttcga tacgccattg agcctttaat agcctatagt gagtcgtatt
5701 aaccgggatc gatatccccc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
5761 gcgtat
//
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