质粒名称: | pCAG-D12L |
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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 |
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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 |
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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 |
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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
pCAGD12L载体基本信息
pCAGD12L载体图谱质粒图谱和多克隆位点信息
pCAGD12L载体简介
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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