质粒名称: | T7opt in pCAGGS |
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出品公司: | 爱迪基因 |
目录编号: | 65974 |
存储实验室: | Benhur Lee |
载体骨架基本信息 |
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载体名称: | pCAGGS |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 4729 bp |
完整质粒大小: | 7381 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
筛选标记: | -- |
高拷贝/低拷贝: | 高拷贝 |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | T7-opt,T7 RNA polymerase |
物种来源: | bacteriophage |
插入大小: | 2652 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | T7opt in pCAGGS was a gift from Benhur Lee (Addgene plasmid # 65974 ; http://n2t.net/addgene:65974 ; RRID:Addgene_65974) |
参考文献部分: | Efficient reverse genetics reveals genetic determinants of budding and fusogenic differences between Nipah and Hendra viruses and enables real-time monitoring of viral spread in small animal models of henipavirus infection. Yun T, Park A, Hill TE, Pernet O, Beaty SM, Juelich TL, Smith JK, Zhang L, Wang YE, Vigant F, Gao J, Wu P, Lee B, Freiberg AN. J Virol. 2015 Jan 15;89(2):1242-53. doi: 10.1128/JVI.02583-14. Epub 2014 Nov 12. 10.1128/JVI.02583-14 PubMed 25392218 |
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载体序列
LOCUS T7opt in pCAGGS 7381 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Human codon-optimized T7 polymerase. ACCESSION . VERSION . KEYWORDS T7opt in pCAGGS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7381) AUTHORS Yun T, Park A, Hill TE, Pernet O, Beaty SM, Juelich TL, Smith JK, Zhang L, Wang YE, Vigant F, Gao J, Wu P, Lee B, Freiberg AN TITLE Efficient reverse genetics reveals genetic determinants of budding and fusogenic differences between Nipah and Hendra viruses and enables real-time monitoring of viral spread in small animal models of henipavirus infection. JOURNAL J Virol. 2015 Jan 15;89(2):1242-53. doi: 10.1128/JVI.02583-14. Epub 2014 Nov 12. PUBMED 25392218 REFERENCE 2 (bases 1 to 7381) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7381 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 368..645 /label=chicken beta-actin promoter intron 647..1664 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1588..1610 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1672..1691 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" primer_bind complement(4385..4404) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_site 4450..4505 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal" primer_bind complement(4504..4523) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind complement(4866..4882) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4866..4882) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4879..4901) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4890..4906 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(4914..4944) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4959..4980 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 5039..5234 /label=SV40 promoter /note="SV40 early promoter" rep_origin 5085..5220 /label=SV40 ori /note="SV40 origin of replication" primer_bind 5147..5166 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" polyA_signal 5240..5374 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5277..5296) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5331..5350 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5442..5459) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5613..6201) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5693..5712) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6372..7232) /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 6995..7014 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7233..7337) /gene="bla" /label=AmpR promoter enhancer 7368..366 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" ORIGIN 1 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc 61 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca 121 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt 181 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg 241 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag 301 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt 361 accatggtcg aggtgagccc cacgttctgc ttcactctcc ccatctcccc cccctcccca 421 cccccaattt tgtatttatt tattttttaa ttattttgtg cagcgatggg ggcggggggg 481 gggggggggc gcgcgccagg cggggcgggg cggggcgagg ggcggggcgg ggcgaggcgg 541 agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa agtttccttt tatggcgagg 601 cggcggcggc ggcggcccta taaaaagcga agcgcgcggc gggcggggag tcgctgcgac 661 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac 721 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt 781 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc 841 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg 901 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg 961 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc 1021 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt 1081 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag 1141 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg 1201 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg 1261 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg 1321 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt 1381 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc 1441 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt 1501 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct 1561 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta 1621 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc 1681 tggttattgt gctgtctcat cattttggca aagaattcgc caccatgaac accatcaata 1741 ttgccaagaa cgacttttct gatatcgagc tggccgctat tccattcaat acactggctg 1801 accactacgg agagcggctg gcccgcgaac agctggctct ggagcatgaa agctatgaga 1861 tgggagaagc ccgattcagg aagatgtttg agaggcagct gaaagctggg gaagtggcag 1921 acaacgcagc cgctaagcca ctgattacca cactgctgcc caaaatgatc gccagaatta 1981 atgattggtt cgaggaagtg aaggcaaaac gaggaaagag gcctaccgcc ttccagtttc 2041 tgcaggagat caagccagaa gctgtggcat acatcaccat caagactacc ctggcatgcc 2101 tgacaagcgc cgacaacaca actgtgcagg cagtcgcttc cgcaatcgga cgagctattg 2161 aggacgaagc acgctttggg agaatccggg atctggaggc caagcacttc aagaaaaacg 2221 tggaggaaca gctgaacaag agggtggggc atgtctataa gaaggccttc atgcaggtgg 2281 tcgaggccga catgctgtca aagggactgc tgggaggaga ggcctggagc tcctggcaca 2341 aagaagatag catccatgtg ggagtccgct gcatcgagat gctgattgaa tctactggga 2401 tggtgagtct gcaccgacag aacgccggcg tggtcggaca ggactctgag acaatcgaac 2461 tggctcccga gtatgccgaa gctattgcaa ctagagccgg agctctggca gggatcagtc 2521 caatgttcca gccctgcgtg gtccccccta agccttggac tggcatcacc gggggcggat 2581 actgggctaa tggaaggaga ccactggcac tggtgcgaac acactctaag aaagccctga 2641 tgagatacga ggatgtctat atgcccgaag tgtataaggc catcaacatt gctcagaata 2701 cagcatggaa aattaacaag aaagtgctgg ccgtcgctaa tgtgatcact aagtggaaac 2761 attgtcccgt ggaggacatc cctgccattg aacgggagga actgcctatg aagccagagg 2821 acatcgatat gaaccctgaa gctctgaccg catggaagcg cgcagccgct gcagtgtaca 2881 gaaaggataa agcccggaag tcccggcgca tttctctgga gttcatgctg gaacaggcca 2941 acaagtttgc taatcacaaa gcaatctggt tcccctacaa catggactgg cgcggacgag 3001 tctatgccgt gagcatgttc aaccctcagg ggaatgatat gacaaagggc ctgctgactc 3061 tggctaaggg gaaaccaatt gggaaggagg gctactattg gctgaaaatc cacggggcca 3121 attgcgctgg cgtcgacaag gtgccattcc ccgagaggat caagttcatc gaggaaaacc 3181 atgaaaatat tatggcatgt gccaagtctc ccctggagaa cacatggtgg gccgaacagg 3241 atagtccttt ctgctttctg gccttctgtt ttgagtacgc tggagtgcag caccatgggc 3301 tgagttataa ttgctccctg ccactggcct ttgacggctc ttgtagtgga atccagcact 3361 tctccgcaat gctgagggat gaggtcggag gaagagcagt gaacctgctg ccatctgaga 3421 cagtgcagga catctacggc attgtcgcca agaaagtgaa tgagatcctg caggctgacg 3481 caattaacgg gactgataat gaggtggtca ccgtcacaga tgaaaacact ggcgagatca 3541 gcgaaaaggt gaaactggga accaaggccc tggctggaca gtggctggca tacggagtca 3601 cccgctcagt gacaaagcga agcgtgatga ccctggctta tggcagcaaa gagttcggct 3661 tcaggcagca ggtgctggaa gacaccatcc agccagccat tgattccgga aaggggctga 3721 tgtttacaca gcccaaccag gccgctggct acatggccaa gctgatctgg gagtcagtga 3781 gcgtcacagt ggtcgcagcc gtggaagcta tgaattggct gaagtccgct gcaaaactgc 3841 tggccgctga ggtgaaggac aagaaaactg gcgaaattct gaggaaaaga tgcgccgtcc 3901 actgggtgac ccctgatgga ttcccagtgt ggcaggagta taagaaaccc atccagacca 3961 gactgaacct gatgttcctg ggccagtttc ggctgcagcc tacaatcaac actaataagg 4021 acagtgagat tgatgctcat aaacaggaat cagggattgc acctaatttt gtgcacagcc 4081 aggacggctc ccatctgcgg aagactgtgg tctgggctca cgagaaatac ggcatcgaat 4141 ccttcgcact gattcatgac tcttttggaa ccatcccagc cgatgcagcc aacctgttca 4201 aggctgtccg cgagactatg gtggacacct acgaaagttg tgatgtgctg gccgacttct 4261 atgatcagtt tgctgaccag ctgcacgagt cacagctgga taagatgccc gcactgcctg 4321 ccaaaggcaa cctgaatctg agagacatcc tggagtccga tttcgcattt gcctgactcg 4381 aggatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt gagcatctga 4441 cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat tttttgtgtc 4501 tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat gagtatttgg 4561 tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt tggctataaa 4621 gaggtcatca gtatatgaaa cagccccctg ctgtccattc cttattccat agaaaagcct 4681 tgacttgagg ttagattttt tttatatttt gttttgtgtt atttttttct ttaacatccc 4741 taaaattttc cttacatgtt ttactagcca gatttttcct cctctcctga ctactcccag 4801 tcatagctgt ccctcttctc ttatggagat ccctcgacct gcagcccaag cttggcgtaa 4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 4921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc 5041 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 5101 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 5161 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 5221 taggcttttg caaaaagcta acttgtttat tgcagcttat aatggttaca aataaagcaa 5281 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc 5341 caaactcatc aatgtatctt atcatgtctg gatccgctgc attaatgaat cggccaacgc 5401 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 5461 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 5521 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 5581 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 5641 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 5701 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 5761 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 5821 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 5881 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 5941 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 6001 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 6061 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 6121 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 6181 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 6241 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 6301 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 6361 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 6421 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 6481 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 6541 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 6601 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 6661 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 6721 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 6781 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 6841 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 6901 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 6961 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 7021 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 7081 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 7141 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 7201 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 7261 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 7321 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgggtcgac attgattatt 7381 g //
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