| 质粒名称: | pT7-VP6SA11 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89166 |
| 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: | 4430 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 VP6 |
| 物种来源: | Rotavirus SA11 |
| 插入大小: | 1356 bp |
| GenBank编号 | LC178568 |
| 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-VP6SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89166 ; http://n2t.net/addgene:89166 ; RRID:Addgene_89166) |
| 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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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-VP6SA11 4430 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Rotavirus SA11 VP6 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pT7-VP6SA11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4430)
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 4430)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..4430
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(29..46)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(200..788)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(280..299)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(959..1819)
/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 1582..1601
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1820..1924)
/gene="bla"
/label=AmpR promoter
primer_bind 1992..2010
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2048..2070)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2170..2189
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2399..2418
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2708..2726
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2708..2725
/label=SP6
/note="SP6 promoter, forward primer"
terminator 2794..2841
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 2837..2855
/label=T7 Term
/note="T7 terminator, reverse primer"
promoter complement(4356..4374)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
1 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
61 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
121 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
181 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
241 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
301 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
361 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
421 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
481 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
541 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
601 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
661 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
721 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
781 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
841 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
901 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
961 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
1021 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
1081 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
1141 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
1201 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
1261 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
1321 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
1381 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
1441 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
1501 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
1561 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
1621 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
1681 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
1741 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
1801 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
1861 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
1921 cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat
1981 taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg
2041 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
2101 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
2161 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccattcgacg ctctccctta
2221 tgcgactcct gcattaagaa gcagcccagt agtaggttga ggccgttgag caccgccgcc
2281 gcaaggaatg gtgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
2341 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
2401 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
2461 tgcgtccggc gtagaggatc tggctagcga tgaccctgct gattggttcg ctgaccattt
2521 ccgggtgcgg gacggcgtta ccagaaactc agaaggttcg tccaaccaaa ccgactctga
2581 cggcagttta cgagagagat gatagggtct gcttcagtaa gccagatgct acacaattag
2641 gcttgtacat attgtcgtta gaacgcggct acaattaata cataacctta tgtatcatac
2701 acatacgatt taggtgacac tatagaatac aagcttgcat gcctgcaggt cgactctaga
2761 ggatctcgat ccggatatag ttcctccttt cagcaaaaaa cccctcaaga cccgtttaga
2821 ggccccaagg ggttatgcta gttattgctc agcggtggca gcagccaact cagcttcctt
2881 tcgggctttg ttagcagccg gatccttttt tttgagctct cccttagcca tccgagtgga
2941 cgacgtcctc cttcggatgc ccaggtcgga ccgcgaggag gtggagatgc catgccgacc
3001 cggtcacatc ctctcactat accatctgag tgattactct gtgtagcgaa tgcgtatacg
3061 catccttact tacagagttc aaacagcttg atatagctac atactcacca aagttggata
3121 ccaagttgtt agcttggtcc tcatttaatg agcatgcttc taatggaagc cactgtaaat
3181 acgcgttgca aattgtcctc cctagacggt gaataattgg tgattaaatc agtccagttc
3241 atacctggtg gaaagactgg tccaactggt attgcgtact cttgcctaac ggatgttaca
3301 tttgctaata gagtttcact tgcatcggcg agtacagact cacaaactgc agactcaatt
3361 ctaagtgtca atcccactgt tgcatgatgt tcgaatggct gtgcattcgg gaatagtact
3421 gctactgctg gtgtcatgtt tggtggtctc attaactgga atgatagtct aatagtatca
3481 aaatttctag ctacgatagt tccaaatctt gcttgataag tgtttattat ctgtccattc
3541 aatagaaact ccacttcaac gttattcggc ctgagaatca ctgggttgaa aaaccatgta
3601 gttgccccgt cagctgaatt gatcactctt ggaaaactaa acctttccgc gtctggtaga
3661 agagttatcg tagctgtagt taacactctt cggagtggca caatatgctc aaattgttgt
3721 atattggctg gtgcgttaat agcacatgag tagtcaaatc cagcgacttg aatttccgat
3781 cctgcgttta accacattgt gcccatcaaa ttatcatgag cgggttgtga tctatttagt
3841 gtaaatgatg ctgaataagg aaaaatgttt ggtttatgaa aagtgaaacc tgtcctctgt
3901 cttctatttt gcaaattcca gttttctatg tattccgacg aattatcaaa atttattctt
3961 ttgaatttaa tggctgacag ctttcttagc gagtctgatt gaggtgcaat tccgttcctt
4021 tgtgattctc taaccatctc atccatgcat acattgtcta caaaatccac gaaataatca
4081 attgtatttc ttgccgtttc aacataatta gcgtctaagt tcagcaaagt tgttccaagt
4141 aacccgaaat taaaattcca gtttctaatt ggcaaattac cgattcctcc agtttgaaat
4201 tcatttccat tcatagtaat tatcatttga ttaaattgtt gaattagatc actcacgtta
4261 gaatacaatg tgccttcgac aattttgtct ctagcgtctt taagagtctt tgacaaagag
4321 tataggacat ccatgttgaa gacttcgttt aaaagcctat agtgagtcgt attaacccgg
4381 gatcgatatc cccctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
//
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