| 质粒名称: | pCAG-D12L |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 89161 |
| 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: | 6577 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 D12L |
| 插入大小: | 864 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-D12L was a gift from Takeshi Kobayashi (Addgene plasmid # 89161 ; http://n2t.net/addgene:89161 ; RRID:Addgene_89161) |
| 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
pCAG-D12L载体 pCAGD12L质粒基本信息
pCAG-D12L质粒图谱 pCAGD12L载体图谱和pCAG-D12L载体序列 pCAGD12L质粒序列多克隆位点信息
pCAG-D12L质粒 pCAGD12L载体简介
pCAG-D12L质粒序列 pCAGD12L载体序列
LOCUS pCAG-D12L 6577 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Express vaccinia virus D12L in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS pCAG-D12L
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6577)
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 6577)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..6577
/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(2722..2741)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 2787..2842
/label=-beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(2841..2860)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(3194..3214)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
primer_bind 3441..3460
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3621..4220
/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(3621..3640)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 4117..4137
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
polyA_signal 4450..4584
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4487..4506)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4541..4560
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4652..4669)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4823..5411)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4903..4922)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(5582..6442)
/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 6205..6224
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6443..6547)
/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 ctatggatga aattgtaaaa aatatccggg agggaacgca
1801 tgtccttctt ccattttatg aaacattgcc agaacttaat ctgtctctag gtaaaagccc
1861 attacctagt ctggaatacg gagctaatta ctttcttcag atttctagag ttaatgatct
1921 aaatagaatg ccgaccgaca tgttaaaact ttttacacat gatatcatgt taccagaaag
1981 cgatctagat aaagtctatg aaattttaaa gattaatagc gtaaagtatt atgggaggag
2041 tactaaagcg gacgccgtag ttgccgacct cagcgcacgc aataaactgt tcaaacgtga
2101 acgagatgct attaaatcta ataatcatct cactgaaaac aatctataca ttagcgatta
2161 taagatgtta accttcgacg tgtttcgacc attatttgat tttgtaaacg aaaaatattg
2221 tattattaaa cttccaactt tattcggtag aggtgtaatc gatactatga gaatatattg
2281 tagtctcttt aaaaatgtta gactgctaaa atgcgtaagc gatagctggt tgaaagatag
2341 cgccattatg gtggctagtg atgtttgtaa aaaaaatttg gatttattta tgtctcatgt
2401 taagtccgtc actaagtctt cttcttggaa ggatgtgaac agtgttcaat ttagtatttt
2461 aaacaatcca gtggatacgg aattcattaa taagttctta gagttttcga atagagtata
2521 cgaagctctc tattacgttc actcgttgct ttattctagt atgacttctg attcaaaaag
2581 tatcgaaaac aaacatcaga gaagactagt taaactactg ctgtgaagat cttcgaattc
2641 actcctcagg tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct
2701 cacaaatacc actgagatcg atctttttcc ctctgccaaa aattatgggg acatcatgaa
2761 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
2821 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
2881 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
2941 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
3001 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
3061 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
3121 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
3181 gcccaagctc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
3241 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
3301 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
3361 ccgcccctaa ctccgcccat cccgccccta actccgccca gtttccgccc ttctccgccc
3421 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
3481 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gcttgcatgc
3541 ctgcaggtcg gccgccacga ccggtgccgc caccatcccc tgacccacgc ccctgacccc
3601 tcacaaggag acgaccttcc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
3661 acgacgtccc ccgggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
3721 gccacaccgt cgacccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
3781 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
3841 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
3901 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
3961 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
4021 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
4081 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
4141 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
4201 cccgcaagcc cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg
4261 cacgacccca tggctccgac cgaagccacc cggggcggcc ccgccgaccc cgcacccgcc
4321 cccgaggccc accgactcta gaggatcata atcagccata ccacatttgt agaggtttta
4381 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
4441 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
4501 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
4561 aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag
4621 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
4681 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
4741 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
4801 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
4861 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
4921 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
4981 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
5041 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
5101 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
5161 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
5221 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
5281 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
5341 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
5401 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
5461 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
5521 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
5581 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
5641 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
5701 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
5761 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
5821 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
5881 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
5941 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
6001 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
6061 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
6121 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
6181 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
6241 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
6301 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
6361 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
6421 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
6481 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
6541 ttccgcgcac atttccccga aaagtgccac ctgggtc
//
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