| 质粒名称: | pCAG-D1R |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89160 |
| 存储实验室: | Takeshi Kobayashi |
|
载体骨架基本信息 |
|
| 载体名称: | pCAGGS |
| 空载体大小: | 5713 bp |
| 完整质粒大小: | 8248 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 嘌呤霉素 |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
|
插入基因信息 |
|
| 插入基因名称: | Vaccinia virus D1R |
| 物种来源: | Vaccinia virus |
| 插入大小: | 2535 bp |
| 启动子: | CAG promotor |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | CTAGAGCCTCTGCTAACCATGTTC |
| 3’测序引物 | TCAGTGGTATTTGTGAGCCAGG |
|
文献引用方法 |
|
| 材料和方法部分: | pCAG-D1R was a gift from Takeshi Kobayashi (Addgene plasmid # 89160 ; http://n2t.net/addgene:89160 ; RRID:Addgene_89160) |
| 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 pCAG-D1R 8248 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express vaccinia virus D1R in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCAG-D1R
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8248)
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 8248)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8248
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..657
/label=chicken beta-actin promoter
intron 658..1672
/label=chimeric intron
/note="chimera between introns from chicken beta-actin and
rabbit beta-globin"
primer_bind 1596..1618
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1680..1699
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
primer_bind complement(4393..4412)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 4458..4513
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(4512..4531)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(4865..4885)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind 5112..5131
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 5292..5891
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(5292..5311)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 5788..5808
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 6121..6255
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6158..6177)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 6212..6231
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(6323..6340)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6494..7082)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(6574..6593)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7253..8113)
/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 7876..7895
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8114..8218)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
481 tgggggcggg gggggggggg gggggcgcca ggcggggcgg ggcggggcga ggggcggggc
541 ggggcgaggc ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct
601 tttatggcga ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga
661 gtcgctgcgt tgccttcgcc ccgtgccccg ctccgcgccg cctcgcgccg cccgccccgg
721 ctctgactga ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc
781 tgtaattagc gcttggttta atgacggctc gtttcttttc tgtggctgcg tgaaagcctt
841 aaagggctcc gggagggccc tttgtgcggg ggggagcggc tcggggggtg cgtgcgtgtg
901 tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg agcgctgcgg
961 gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg ccgggggcgg
1021 tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg gtgtgtgcgt
1081 gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc tgcacccccc
1141 tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg ggcgtggcgc
1201 ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg gggcggggcc
1261 gcctcgggcc ggggagggct cgggggaggg gcgcggcggc ccccggagcg ccggcggctg
1321 tcgaggcgcg gcgagccgca accattgcct tttatggtaa tcgtgcgaga gggcgcaagg
1381 acttcctttg tcccaaatct gtgcggagcc gaaatctggg aggcgccgcc gcaccccctc
1441 tagcgggcgc ggggcgaagc ggtgcggcgc cggcaggaag gaaatgggcg gggagggcct
1501 tcgtgcgtcg ccgcgccgcc gtccccttct ccctctccag cctcggggct gtccgcgggg
1561 ggacggctgc cttcgggggg gacggggcag ggcggggttc ggcttctggc gtgtgaccgg
1621 cggctctaga gcctctgcta accatgttca tgccttcttc tttttcctac agctcctggg
1681 caacgtgctg gttattgtgc tgtctcatca ttttggcaaa gaattaatta agtttaaacg
1741 gcgcgccgat atcttaagat ctatggatgc caacgtagta tcatcttcta ctattgcaac
1801 gtatatagac gctttagcga agaatgcttc agaattagaa cagaggtcta ccgcatacga
1861 aataaataat gaattggaac tagtatttat taagccgcca ttaattactt tgacaaatgt
1921 agtgaatatc tctacgattc aggaatcgtt tattcgattt accgttacta ataaggaagg
1981 tgttaaaatt agaactaaga ttccattatc taaggtacat ggtctagatg taaaaaatgt
2041 acagttagta gatgctatag ataacatagt ttgggaaaag aaatcattag tgacggaaaa
2101 tcgtcttcac aaagaatgct tgttgagact atcgacagag gaacgttata tatttttgga
2161 ttacaagaaa tatggatcct ctatccgact agaattagtc aatcttattc aagcaaaaac
2221 aaaaaacttt acgatagact ttaagctaaa atattttcta ggatccggtg cccagtctaa
2281 aagttcttta ttacacgcta ttaatcatcc aaagtcaagg cctaatacat ctctggaaat
2341 agaattcaca cctagagaca atgaaaaagt tccatatgat gaactaataa aggaattgac
2401 aactctatca cgtcatatat ttatggcttc tccagagaat gtaattcttt ctccgcctat
2461 taacgcgcct ataaaaacct ttatgttgcc taaacaagat atagtaggtt tggatctgga
2521 aaatctatat gccgtaacta agactgacgg cattcctata actatcagag ttacatcaaa
2581 cgggttgtat tgttatttta cacatcttgg ttatattatt agatatcctg ttaagagaat
2641 aatagattcc gaagtagtag tctttggtga ggcagttaag gataagaact ggaccgtata
2701 tctcattaag ctaatagagc ctgtgaatgc aatcaatgat agactagaag aaagtaagta
2761 tgttgaatct aaactagtgg atatttgtga tcggatagta ttcaagtcaa agaaatatga
2821 aggtccgttt actacaacta gtgaagtcgt cgatatgtta tctacatatt taccaaagca
2881 accagaaggt gttattctgt tctattcaaa gggacctaaa tctaacattg attttaaaat
2941 taaaaaggaa aatactatag accaaactgc aaatgtagta tttaggtaca tgtccagtga
3001 accaattatc tttggagaat cgtctatctt tgtagagtat aagaaattta gcaacgataa
3061 aggctttcct aaagaatatg gttctggtaa gattgtgtta tataacggcg ttaattatct
3121 aaataatatc tattgtttgg aatatattaa tacacataat gaagtgggta ttaagtccgt
3181 ggttgtacct attaagttta tagcagaatt cttagttaat ggagaaatac ttaaacctag
3241 aattgataaa accatgaaat atattaactc agaagattat tatggaaatc aacataatat
3301 catagtcgaa catttaagag atcaaagcat caaaatagga gatatcttta acgaggataa
3361 actatcggat gtgggacatc aatacgccaa taatgataaa tttagattaa atccagaagt
3421 tagttatttt acgaataaac gaactagagg accgttggga attttatcaa actacgtcaa
3481 gactcttctt atttctatgt attgttccaa aacattttta gacgattcca acaaacgaaa
3541 ggtattggcg attgattttg gaaacggtgc tgacctggaa aaatactttt atggagagat
3601 tgcgttattg gtagcgacgg atccggatgc tgatgctata gctagaggaa atgaaagata
3661 caacaaatta aactctggaa ttaaaaccaa gtactacaaa tttgactaca ttcaggaaac
3721 tattcgatcc gatacatttg tctctagtgt cagagaagta ttctattttg gaaagtttaa
3781 tatcatcgac tggcagtttg ctatccatta ttcttttcat ccgagacatt atgctaccgt
3841 catgaataac ttatccgaac taactgcttc tggaggcaag gtattaatca ctaccatgga
3901 cggagacaaa ttatcaaaat taacagataa aaagactttt ataattcata agaatttacc
3961 tagtagcgaa aactatatgt ctgtagaaaa aatagctgat gatagaatag tggtatataa
4021 tccatcaaca atgtctactc caatgactga atacattatc aaaaagaacg atatagtcag
4081 agtgtttaac gaatacggat ttgttcttgt agataacgtt gatttcgcta caattataga
4141 acgaagtaaa aagtttatta atggcgcatc tacaatggaa gatagaccgt ctacaaaaaa
4201 ctttttcgaa ctaaatagag gagccattaa atgtgaaggt ttagatgtcg aagacttact
4261 tagttactat gttgtttatg tcttttctaa gcggtaaaga tcttcgaatt cactcctcag
4321 gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac
4381 cactgagatc gatctttttc cctctgccaa aaattatggg gacatcatga agccccttga
4441 gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt gttggaattt
4501 tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac atcagaatga
4561 gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga acaaaggttg
4621 gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct tattccatag
4681 aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat ttttttcttt
4741 aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc tctcctgact
4801 actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc agcccaagct
4861 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
4921 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
4981 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
5041 actccgccca tcccgcccct aactccgccc agtttccgcc cttctccgcc ccatggctga
5101 ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag
5161 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agcttgcatg cctgcaggtc
5221 ggccgccacg accggtgccg ccaccatccc ctgacccacg cccctgaccc ctcacaagga
5281 gacgaccttc catgaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc
5341 cccgggccgt acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg
5401 tcgacccgga ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg
5461 tcgggctcga catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga
5521 ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg
5581 agttgagcgg ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc
5641 ggcccaagga gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca
5701 agggtctggg cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc
5761 ccgccttcct ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca
5821 ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac ctggtgcatg acccgcaagc
5881 ccggtgcctg acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc
5941 atggctccga ccgaagccac ccggggcggc cccgccgacc ccgcacccgc ccccgaggcc
6001 caccgactct agaggatcat aatcagccat accacatttg tagaggtttt acttgcttta
6061 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
6121 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
6181 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
6241 tatcatgtct ggatccgctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
6301 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
6361 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
6421 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
6481 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
6541 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
6601 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
6661 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
6721 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
6781 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
6841 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
6901 tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
6961 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
7021 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
7081 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
7141 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
7201 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
7261 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
7321 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
7381 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
7441 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
7501 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
7561 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
7621 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
7681 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
7741 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
7801 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
7861 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
7921 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
7981 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
8041 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
8101 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
8161 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
8221 catttccccg aaaagtgcca cctgggtc
//
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