| 質粒名稱: | pT7-NSP5SA11 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 89172 |
| 存儲實驗室: | Takeshi Kobayashi |
|
載體骨架基本信息 |
|
| 載體名稱: | p3E5 |
| 載體出品公司: | -- |
| 空載體大小: | 3074 bp |
| 完整質粒大小: | 3741 bp |
| 載體修飾: | -- |
| 載體類型: | 哺乳細胞表達 |
| 篩選標記: | -- |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱: | Rotavirus SA11 NSP5 |
| 物種來源: | Rotavirus SA11 |
| 插入大小: | 667 bp |
| 基因突變: | -- |
| GenBank編號 | LC178574 |
| 啟動子: | T7 promotor |
| 標簽/融合蛋白: | -- |
| 克隆方法: | Gibson克隆 |
| 5’克隆位點: | -- |
| 3’克隆位點: | -- |
| 5’測序引物 | T7 forward CTGTGGATAACCGTATTACCG |
| 3’測序引物 | T7 terminal GCTAGTTATTGCTCAGCGG |
|
文獻引用方法 |
|
| 材料和方法部分: | pT7-NSP5SA11 was a gift from Takeshi Kobayashi (Addgene plasmid # 89172 ; //n2t.net/addgene:89172 ; RRID:Addgene_89172) |
| 參考文獻部分: | 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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pT7-NSP5SA11載體 pT7NSP5SA11質粒基本信息
pT7-NSP5SA11質粒圖譜 pT7NSP5SA11載體圖譜和pT7-NSP5SA11載體序列 pT7NSP5SA11質粒序列多克隆位點信息
pT7-NSP5SA11質粒 pT7NSP5SA11載體簡介
pT7-NSP5SA11質粒序列 pT7NSP5SA11載體序列
LOCUS pT7-NSP5SA11 3740 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express Rotavirus SA11 NSP5 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pT7-NSP5SA11
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3740)
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 3740)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
//www.snapgene.com
FEATURES Location/Qualifiers
source 1..3740
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 33..44
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
promoter complement(90..108)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(192..209)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(363..951)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(443..462)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(1122..1982)
/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 1745..1764
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1983..2087)
/gene="bla"
/label=AmpR promoter
primer_bind 2155..2173
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(2211..2233)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 2333..2352
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 2562..2581
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
promoter 2871..2889
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 2871..2888
/label=SP6
/note="SP6 promoter, forward primer"
terminator 2957..3004
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
primer_bind 3000..3018
/label=T7 Term
/note="T7 terminator, reverse primer"
ORIGIN
1 aagacgattc attcttatat atagttgaag gaatagaagg aagactcgtc acgtcaatac
61 tgagagacat cactgtagcg ctttaaaagc ctatagtgag tcgtattaac cgggatcgat
121 atccccctgc attaatgaat cggccaatgc gcggggagag gcggtttgcg tattgggcgc
181 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
241 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
301 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
361 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
421 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
481 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
541 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
601 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
661 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
721 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
781 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
841 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
901 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
961 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
1021 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
1081 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
1141 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
1201 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
1261 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
1321 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
1381 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
1441 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
1501 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
1561 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
1621 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
1681 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
1741 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
1801 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
1861 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
1921 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
1981 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
2041 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
2101 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
2161 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
2221 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
2281 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
2341 gagcagattg tactgagagt gcaccattcg acgctctccc ttatgcgact cctgcattaa
2401 gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga atggtgcaag
2461 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
2521 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
2581 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
2641 atctggctag cgatgaccct gctgattggt tcgctgacca tttccgggtg cgggacggcg
2701 ttaccagaaa ctcagaaggt tcgtccaacc aaaccgactc tgacggcagt ttacgagaga
2761 gatgataggg tctgcttcag taagccagat gctacacaat taggcttgta catattgtcg
2821 ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg atttaggtga
2881 cactatagaa tacaagcttg catgcctgca ggtcgactct agaggatctc gatccggata
2941 tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca aggggttatg
3001 ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct ttgttagcag
3061 ccggatcctt ttttttgagc tctcccttag ccatccgagt ggacgacgtc ctccttcgga
3121 tgcccaggtc ggaccgcgag gaggtggaga tgccatgccg acccggtcac aaaacgggag
3181 tggggagctc cctagtgtgt tcccaggtca gacttacaaa tcttcaatca attgcattgc
3241 gacttgtttc attctcattc ttaatgcgaa gtatttcttc ttatatttac agttcttaca
3301 tttaccatca tcactatctg aatcatccag tacataatca tctgaatcag actctgcttc
3361 aattcttggg tagtgcttcc tacttttatg gttttgtttt gatttctcct ttttagtatc
3421 cgtagatatt gatattgatg agtccaaatt agcttttact tttaagcctt tatttaagga
3481 gaaatccact tgatcgcatc cgacattact tgaaggtcgt gattgtgctg atgaatccat
3541 agacacgcca gcgtctgcgt ttgtcttaac tgcattcgat ctaatcgaaa aactggtgag
3601 tggatcgttt gaagcagaat cagatggtcc aatatcctct ggagacttcg acagcatgta
3661 tttattgaat gcttctgcat ctggtgaaat gtactgttca ctcctaccaa tagattttcc
3721 agaaagagtt gacgttgttg
//
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