| 质粒名称: | pT7-VP1SA11 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89162 |
| 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: | 6376 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 VP1 |
| 插入大小: | 3302 bp |
| GenBank编号 | LC178564 |
| 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-VP1SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89162 ; http://n2t.net/addgene:89162 ; RRID:Addgene_89162) |
| 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
pT7VP1SA11载体基本信息
pT7VP1SA11载体图谱质粒图谱和多克隆位点信息
pT7VP1SA11载体简介
pT7VP1SA11载体序列
LOCUS pT7-VP1SA11 6376 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Rotavirus SA11 VP1 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pT7-VP1SA11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6376)
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 6376)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6376
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(22..44)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 144..163
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 373..392
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 682..700
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 682..699
/label=SP6
/note="SP6 promoter, forward primer"
terminator 768..815
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 811..829
/label=T7 Term
/note="T7 terminator, reverse primer"
promoter complement(4276..4294)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4379..4396)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4550..5138)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4630..4649)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5309..6169)
/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 5932..5951
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6170..6274)
/gene="bla"
/label=AmpR promoter
primer_bind 6342..6360
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
ORIGIN
1 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
61 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
121 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattc
181 gacgctctcc cttatgcgac tcctgcatta agaagcagcc cagtagtagg ttgaggccgt
241 tgagcaccgc cgccgcaagg aatggtgcaa ggagatggcg cccaacagtc ccccggccac
301 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
361 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
421 gatgccggcc acgatgcgtc cggcgtagag gatctggcta gcgatgaccc tgctgattgg
481 ttcgctgacc atttccgggt gcgggacggc gttaccagaa actcagaagg ttcgtccaac
541 caaaccgact ctgacggcag tttacgagag agatgatagg gtctgcttca gtaagccaga
601 tgctacacaa ttaggcttgt acatattgtc gttagaacgc ggctacaatt aatacataac
661 cttatgtatc atacacatac gatttaggtg acactataga atacaagctt gcatgcctgc
721 aggtcgactc tagaggatct cgatccggat atagttcctc ctttcagcaa aaaacccctc
781 aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt ggcagcagcc
841 aactcagctt cctttcgggc tttgttagca gccggatcct tttttttgag ctctccctta
901 gccatccgag tggacgacgt cctccttcgg atgcccaggt cggaccgcga ggaggtggag
961 atgccatgcc gacccggtca catctaagcg ttctaatctt gaaagaagtt cgcgttagtg
1021 tacggcgaac gtaatgacgt gatgttccac atcttcttcc ataattttat catttctgat
1081 ttagggtaat tgcaaaatag gcttatccat gaaccatttt ttatagcgta gtttataact
1141 tctagctttg catataagtg taatatgaat gtaacagctg gtatttttcc cttaaatggt
1201 attcttatca gcttctccaa gtccggtgaa ttgaaatcaa ataattgata gcatccataa
1261 ttaatggaca ataaattgta cacgtatgat tctagtattc tatacttatc ttgagaataa
1321 atctttgatc caacatacgt gtcagcgtct attttcggta ttcctaatga aatcagatat
1381 aattgtattt cgttttcatg caatgaaatc actttataca attcttcaat tgatggctta
1441 taaactgaat actttgatat tagtctagat atggctgact tcgaaccgtc atcttcaatt
1501 tgataagttc tggatcctat acattgaatt atatactgta cgttatctgg caaagttggc
1561 ataaattttt ggtattgtat aggtaagtgt gcatcactta ctgtaaaaaa tggttttata
1621 tctctgagta tgtcatttat tgttatttta cttgatttga aggtgaccgg tttctgcaac
1681 atagtcaata aagctgatat ctgtgcacgt cttttctcaa gcgatattgg agcgtatgaa
1741 ttcaatttcg ctttttcagt caaagctatt cctctggaca ctatattatt ctttgaaaat
1801 aataactgtt cagatagtct cgtgacgtaa tttttaaatg ttgatgatgc cgctatctca
1861 tcggctattc ctgatttctg acttgataaa gacatgaacg ctctttgtat atatacttcg
1921 tcatcagtcg ttccgtactc tataataaaa tcctccgagt caaatacttt aaatgttgaa
1981 ttcgtagtta atacgcgttc agaaggaaat agtcttaatg atccagtaat tgcgactgac
2041 gtcatctgca ttatcttagt taaaataaac tccctatcag tttcaaatcc tctaagtcta
2101 tttacaatat aatttgaata taaaattgct gcttgatccc actgcgtact ctgccctctc
2161 ttttcattat taagtagatt tattccagct cgaaaaaata tttttccacc tgcaatgtac
2221 cttttagcaa tttctattcc tactgtggat accagagctt taactttagc attcatgcgt
2281 gcataagttt ctcttacatc gttcgatacg tcttggatca tctgcttagt cacctctgta
2341 ttaaattgta gcaccgcata gttatcatct ccatcaactc ttattatttt tgttgcgaat
2401 gaatgtttgt tagaaatacg tgacaaaacc gttttaatca gtgccaaatt tgctattgaa
2461 tttgctgctt tcgtttgttt ctctcctgat gctactgccc cgtattgtat tttcttaatg
2521 acgttgccat ctggtatttg aacgtatgaa tccatgagat tgatttgtgt ttgtttgtat
2581 aagtttaatg tctgaagaac tttagcatca ttagtcatgt tagctaatat gtccagtccc
2641 attattattc ctttcctaaa tggctgtgta ttatgctgag acgaatccca ctgagataca
2701 tccgtataca agaccattgt gttattagac aaaaaacgcg ttacatcgcc gtatgacaat
2761 aattggtttg attgtgaata aaattcagcg tattctctcg tatgttttgc gtaaatcaac
2821 attttttcaa ctacagcgtg ctgtgctatg aaatattcgt atggcagaat gaatattatt
2881 ctagtccttc ttcctggaac atctcttctt cctaatggta ttggtttatc aacattcact
2941 ggtggtatta taccaggcgt gtatctttcg ttagccatat catccatgac atgcatattc
3001 tttttagttg aaaaaattgt tttcctacca aactttagct gccttgactc accattcgat
3061 gctgacgaca ttgatagtaa accagctaac tcagaatctc gtagcaagtg atcatcatgt
3121 ttaactggtt cttcaagcat tttaactact tcatctctta ttaacattgt atattcccga
3181 tacatttcat cgtccacatt ttctgtatac tcagttttca attgatccag cgcagcatct
3241 agcatttttt gctttctaaa tccaacatga aacgaccatg aatatatctt agccattaat
3301 ggaaattttt cgatagaacg atcaactaac cagtcctgta tcatcttagg tatgtcaact
3361 aatccagctt tcctcatatt gtcaagcatt tgatttaatt cgtcaaatgt ttgatccgga
3421 actattgctc gtacatattt atttgaaaat tccaattcta attcttcatt tgtaataaat
3481 gttccattaa tatccaccaa ggctaccaaa atcgacattg gtgatgacac taactttgct
3541 agagtagatc tattagaata tgataacact attaactctt ttgctatcaa atagtcgtca
3601 tgagagaaaa ctgacattaa catttcaatt gaagaatttg cccatgtcac taagtagtac
3661 ggtttatctt tcattacgtc gtattcatat ctgtactttg cgttatgcct tggtacgcca
3721 tatttattta tcgttgaggc tactatggtg aataggtcta atctcctttt ataaacttta
3781 agtttttcag cgacatcatt ttcatgtttt tccaaccaga acatgactgc atttaaattt
3841 gatgatagcg aagttaaaat tgctggatcc attagtgagt ccgtatattc ctccgctgtt
3901 ggaaataatt cagatgttat tttattgttc tcataatcca attcattcac tgttaagtct
3961 gcctccaatg gtttgccttt cgcatacttc accaattttc tttcaacagc gttatacttg
4021 tcgtaagaat atgatagtat tgacagtaat gtggcatttt ctatgacatc attatattca
4081 acaaacaact ttcttaacga taacccattc tttgagttct ctaaacactt ggaatgaaat
4141 tcaatacatc tattttctaa ttcactgttg gaagagtaat atattggaat ttgaactgca
4201 gattgtgaat tatatataaa tgatagatat tctgacaaga ttagattgta cttccccatt
4261 gtacagcttt aatagcctat agtgagtcgt attaacccgg gatcgatatc cccctgcatt
4321 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
4381 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
4441 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
4501 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
4561 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
4621 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
4681 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
4741 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
4801 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
4861 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
4921 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
4981 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
5041 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
5101 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
5161 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
5221 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
5281 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
5341 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
5401 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
5461 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
5521 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
5581 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
5641 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
5701 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
5761 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
5821 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
5881 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
5941 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
6001 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
6061 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
6121 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
6181 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
6241 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
6301 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
6361 cctttcgtct cgcgcg
//
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