质粒名称: | pGD-p19<质粒图谱.png/td> |
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出品公司: | Addgene |
目录编号: | 192326 |
存储实验室: | Zhenghe Li |
载体骨架基本信息 |
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载体名称: | pGD |
载体出品公司: | Clontech |
空载体大小: | 4038 bp |
完整质粒大小: | 77.2bp |
载体类型: | 植物表达 |
载体抗性: | Kanamycin |
生长温度: | 37 ℃ |
宿主菌: | DH5a |
拷贝数: | 高拷贝 |
插入基因信息 |
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插入基因名称: | tobacco etch virus P1/HC-Pro / TEV P1/HC-Pro |
物种来源: | tobacco etch virus |
启动子: |
duplicated cauliflower mosaic virus 35S promoter
Cloning Information
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克隆方法: | Ligation Independent Cloning |
3’克隆位点: | NotI (未破坏) |
5’测序引物 | 35S promoter/forward primer: CTATCCTTCGCAAGACCCTTC |
3’测序引物 | Nos terminator/reverse primer: TCATCGCAAGACCGGCAA |
文献引用方法 |
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材料和方法部分: |
pGD-P1/HC-Pro was a gift from Zhenghe Li (Addgene plasmid # 196328 ; http://n2t.net/addgene:196328 ; RRID:Addgene_196328)
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参考文献部分: | Construction of a Sonchus Yellow Net Virus minireplicon: a step toward reverse genetic analysis of plant negative-strand RNA viruses. Ganesan U, Bragg JN, Deng M, Marr S, Lee MY, Qian S, Shi M, Kappel J, Peters C, Lee Y, Goodin MM, Dietzgen RG, Li Z, Jackson AO. J Virol. 2013 Oct;87(19):10598-611. doi: 10.1128/JVI.01397-13. Epub 2013 Jul 24. 10.1128/JVI.01397-13 PubMed 23885070 |
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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 pGD-p19 7727 bp ds-DNA circular SYN 12-APR-2023 DEFINITION 35S promoter-driven expression of the tomato bushy stunt virus p19 RNA silencing suppressor. ACCESSION . VERSION . KEYWORDS pGD-p19 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7727) AUTHORS Ganesan U, Bragg JN, Deng M, Marr S, Lee MY, Qian S, Shi M, Kappel J, Peters C, Lee Y, Goodin MM, Dietzgen RG, Li Z, Jackson AO TITLE Construction of a Sonchus Yellow Net Virus minireplicon: a step toward reverse genetic analysis of plant negative-strand RNA viruses. JOURNAL J Virol. 2013 Oct;87(19):10598-611. doi: 10.1128/JVI.01397-13. Epub 2013 Jul 24. PUBMED 23885070 REFERENCE 2 (bases 1 to 7727) AUTHORS . TITLE Direct Submission JOURNAL Exported Apr 12, 2023 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..7727 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 5..29 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" promoter 179..524 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" primer_bind 467..487 /label=35S promoter /note="CaMV 35S promoter, forward primer" terminator 1178..1430 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature 1452..1476 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 2777..3406 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 3835..4908 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 4974..5168 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(5404..5426) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 5512..5652 /label=bom /note="basis of mobility region from pBR322" primer_bind 5567..5586 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(5667..5684) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5838..6426) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5918..5937) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6513..7307) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" ORIGIN 1 ctggtggcag gatatattgt ggtgtaaaca aattgacgct tagacaactt aataacacat 61 tgcggacgtt tttaatgtac tgaattaacg ccgaattaat acgcgtcatg gtggagcacg 121 acactcttgt ctactccaaa aatatcaaag atacagtctc agaagaccaa agggcaattg 181 agacttttca acaaagggta atatccggaa acctcctcgg attccattgc ccagctatct 241 gtcactttat tgtgaagata gtggaaaagg aaggtggctc ctacaaatgc catcattgcg 301 ataaaggaaa ggccatcgtt gaagatgcct ctgccgacag tggtcccaaa gatggacccc 361 cacccacgag gagcatcgtg gaaaaagaag acgttccaac cacgtcttca aagcaagtgg 421 attgatgtga tatctccact gacgtaaggg atgacgcaca atcccactat ccttcgcaag 481 acccttcctc tatataagga agttcatttc atttggagag gacacgctga aatcaccagt 541 ctctctctac aaatctatct ctggatcccc agtctctctc tacaagatct cgagccatgg 601 aacgagctat acaaggaaac gacgctaggg aacaagctaa cagtgaacgt tgggatggag 661 gatcaggagg taccacttct cccttcaaac ttcctgacga aagtccgagt tggactgagt 721 ggcggctaca taacgatgag acgaattcga atcaagataa tccccttggt ttcaaggaaa 781 gctggggttt cgggaaagtt gtatttaaga gatatctcag atacgacagg acggaagctt 841 cactgcacag agtccttgga tcttggacgg gagattcggt taactatgca gcatctcgat 901 ttttcggttt cgaccagatc ggatgtacct atagtattcg gtttcgagga gttagtatca 961 ccgtttctgg agggtcgcga actcttcagc atctctgtga gatggcaatt cggtctaagc 1021 aagaactgct acagcttgcc ccaatcgaag tggaaagtaa tgtatcaaga ggatgccctg 1081 aaggtactca aaccttcgaa aaagaaagcg agtaactgca gtcgacggta ccgcgggccc 1141 gggatccacc ggatctagat aactgatcgt cgacagagct cgttcaaaca tttggcaata 1201 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt 1261 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt 1321 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg 1381 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattaaa 1441 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag 1501 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca 1561 tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat 1621 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag 1681 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt 1741 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag 1801 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc 1861 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc 1921 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac 1981 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt 2041 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac 2101 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag 2161 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg 2221 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc 2281 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc 2341 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt 2401 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc 2461 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg 2521 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct 2581 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc 2641 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg 2701 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa 2761 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa 2821 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt 2881 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc 2941 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg 3001 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat 3061 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac 3121 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc 3181 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc 3241 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc 3301 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg 3361 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa 3421 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc 3481 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag 3541 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt 3601 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat 3661 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac 3721 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg 3781 ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc 3841 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag 3901 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt 3961 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc 4021 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg 4081 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt 4141 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac 4201 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg 4261 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag 4321 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat 4381 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt 4441 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa 4501 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag 4561 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg 4621 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg 4681 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc 4741 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca 4801 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc 4861 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag 4921 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat 4981 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac 5041 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa 5101 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc 5161 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg 5221 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca 5281 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca 5341 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga 5401 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 5461 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat 5521 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag 5581 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 5641 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 5701 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 5761 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 5821 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 5881 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 5941 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 6001 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 6061 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 6121 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 6181 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 6241 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 6301 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 6361 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 6421 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 6481 cgttaaggga ttttggtcat gcattctagg tactaaaaca attcatccag taaaatataa 6541 tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac 6601 tgttcttccc cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg 6661 tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag 6721 atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc 6781 tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg 6841 caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct 6901 aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc 6961 gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg 7021 acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg 7081 acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca 7141 tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct 7201 cccaccagct tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat 7261 ttttcgatca gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt 7321 cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca 7381 attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt 7441 tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac 7501 aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg 7561 tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt 7621 ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc 7681 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctgg //
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