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基本信息
載體類(lèi)型: | 植物RNAi載體,VIGS載體 |
載體抗性: | Kan |
載體標(biāo)簽: | His |
啟動(dòng)子: | 35S |
篩選標(biāo)記: | / |
表達(dá)水平: | 高拷貝 |
載體大小: | 9663bp |
克隆方法: | 多克隆位點(diǎn),限制性?xún)?nèi)切酶 |
5' 測(cè)序引物: | primer F:5'-GGACATTGTTACTCAAGGAA-3' |
3' 測(cè)序引物: | primer R:5'-AACCTAAAACTTCAGACACG-3' |
備注: | 如有侵權(quán)請(qǐng)產(chǎn)權(quán)持有人盡快聯(lián)系法務(wù)15927229196 |
質(zhì)粒圖譜和多克隆位點(diǎn)信息
注意:
客戶(hù)在我公司選擇構(gòu)建載體服務(wù)項(xiàng)目時(shí),請(qǐng)自行提供母源載體。
如要使用本公司保存的母源載體時(shí),本公司不能保證所有母源載體完全符合客戶(hù)預(yù)期,也不承擔(dān)由此產(chǎn)生的任何責(zé)任。
載體描述
Tobacco rattle virus (TRV) is a bipartite, positive-sense RNA virus. TRV1 (i.e., RNA1) encodes 134 and 194 kDa replicase proteins from the genomic RNA, 29 kDa movement protein, and 16 kDa cysteine-rich protein from subgenomic RNAs. TRV1 can replicate and move systemically without TRV2 (i.e., RNA2). TRV2 encodes coat protein from the genomic and two nonstructural proteins from subgenomic RNAs (A). To develop the TRV-based VIGS vector, a cDNA clone of TRV1 or TRV2 (Ppk20 strain) was placed between a duplicated CaMV 35S promoter (2X35S) and nopaline synthase terminator (NOSt) in an Agrobacterium T-DNA vector. In the TRV2 cDNA construct, the nonessential structural genes were replaced with MCS for cloning the target gene sequences. Further, a self-cleaving ribozyme site was included at the 3' end of TRV2. The TRV1 (AF406990)-based viral vector, pTRV1 (linear plasmid 6.791 kb), is used along with pTRV2 for silencing.
The pTRV2 has three variants. The first one is suitable for conventional cloning (AF406991, linear plasmid 9663 nucleotide). The second variant is a gateway-cloning-compatible vector. The third variant is suitable for ligation-independent cloning. Rz, self-cleaving ribozyme; MCS, multiple cloning site; CP, coat protein; MP, movement protein. These vectors were created by Dr. S.P. Dinesh-Kumar, UC Davis, USA. Conventional cloning-compatible (YL156) and ligation-independent-compatible (YY13TRV2) vector plasmid DNA can be obtained from ABRC, Stock No. CD3-1040 vector name YL156. Ligation-independent-compatible TRV2 vector plasmid DNA is ABRC Stock No. CD3-1042, vector name YY13.
載體序列
ORIGIN
1 ataaaacatt gcacctatgg tgttgccctg gctggggtat gtcagtgatc gcagtagaat
61 gtactaattg acaagttgga gaatacggta gaacgtcctt atccaacaca gcctttatcc
121 ctctccctga cgaggttttt gtcagtgtaa tatttctttt tgaactatcc agcttagtac
181 cgtacgggaa agtgactggt gtgcttatct ttgaaatgtt actttgggtt tcggttcttt
241 aggttagtaa gaaagcactt gtcttctcat acaaaggaaa acctgagacg tatcgcttac
301 gaaagtagca atgaaagaaa ggtggtggtt ttaatcgcta ccgcaaaaac gatggggtcg
361 ttttaattaa cttctcctac gcaagcgtct aaacggacgt tggggttttg ctagtttctt
421 tagagaaaac tagctaagtc tttaatgtta tcattagaga tggcataaat ataatacttg
481 tgtctgctga taagatcatt ttaatttgga cgattagact tgttgaacta caggttactg
541 aatcacttgc gctaatcaac atgggagata tgtacgatga atcatttgac aagtcgggcg
601 gtcctgctga cttgatggac gattcttggg tggaatcagt ttcgtggaaa gatctgttga
661 agaagttaca cagcataaaa tttgcactac agtctggtag agatgagatc actgggttac
721 tagcggcact gaatagacag tgtccttatt caccatatga gcagtttcca gataagaagg
781 tgtatttcct tttagactca cgggctaaca gtgctcttgg tgtgattcag aacgcttcag
841 cgttcaagag acgagctgat gagaagaatg cagtggcggg tgttacaaat attcctgcga
901 atccaaacac aacggttacg acgaaccaag ggagtactac tactaccaag gcgaacactg
961 gctcgacttt ggaagaagac ttgtacactt attacaaatt cgatgatgcc tctacagctt
1021 tccacaaatc tctaacttcg ttagagaaca tggagttgaa gagttattac cgaaggaact
1081 ttgagaaagt attcgggatt aagtttggtg gagcagctgc tagttcatct gcaccgcctc
1141 cagcgagtgg aggtccgata cgtcctaatc cctagggatt taaggacgtg aactctgttg
1201 agatctctgt gaaattcaga gggtgggtga taccatattc actgatgcca ttagcgacat
1261 ctaaataggg ctaattgtga ctaatttgag ggaatttcct ttaccattga cgtcagtgtc
1321 gttggtagca tttgagtttc gcaatgcacg aattacttag gaagtggctt gacgacacta
1381 atgtgttatt gttagataat ggtttggtgg tcaaggtacg tagtagagtc ccacatattc
1441 gcacgtatga agtaattgga aagttgtcag tttttgataa ttcactggga gatgatacgc
1501 tgtttgaggg aaaagtagag aacgtatttg tttttatgtt caggcggttc ttgtgtgtca
1561 acaaagatgg acattgttac tcaaggaagc acgatgagct ttattattac ggacgagtgg
1621 acttagattc tgtgagtaag gttaccgaat tctctagaag gcctccatgg ggatccggta
1681 ccgagctcac gcgtctcgag gcccgggcat gtcccgaaga cattaaacta cggttcttta
1741 agtagatccg tgtctgaagt tttaggttca atttaaacct acgagattga cattctcgac
1801 tgatcttgat tgatcggtaa gtcttttgta atttaatttt ctttttgatt ttattttaaa
1861 ttgttatctg tttctgtgta tagactgttt gagatcggcg tttggccgac tcattgtctt
1921 accatagggg aacggacttt gtttgtgttg ttattttatt tgtattttat taaaattctc
1981 aacgatctga aaaagcctcg cggctaagag attgttgggg ggtgagtaag tacttttaaa
2041 gtgatgatgg ttacaaaggc aaaaggggta aaacccctcg cctacgtaag cgttattacg
2101 cccgtctgta cttatatcag tacactgacg agtccctaaa ggacgaaacg ggagaacgct
2161 agccaccacc accaccacca cgtgtgaatt acaggtgacc agctcgaatt tccccgatcg
2221 ttcaaacatt tggcaataaa gtttcttaag attgaatcct gttgccggtc ttgcgatgat
2281 tatcatataa tttctgttga attacgttaa gcatgtaata attaacatgt aatgcatgac
2341 gttatttatg agatgggttt ttatgattag agtcccgcaa ttatacattt aatacgcgat
2401 agaaaacaaa atatagcgcg caaactagga taaattatcg cgcgcggtgt catctatgtt
2461 actagatcgg gaattaaact atcagtgttt gacaggatat attggcgggt aaacctaaga
2521 gaaaagagcg tttattagaa taacggatat ttaaaagggc gtgaaaaggt ttatccgttc
2581 gtccatttgt atgtgcatgc caaccacagg gttcccctcg ggatcaaagt actttgatcc
2641 aacccctccg ctgctatagt gcagtcggct tctgacgttc agtgcagccg tcttctgaaa
2701 acgacatgtc gcacaagtcc taagttacgc gacaggctgc cgccctgccc ttttcctggc
2761 gttttcttgt cgcgtgtttt agtcgcataa agtagaatac ttgcgactag aaccggagac
2821 attacgccat gaacaagagc gccgccgctg gcctgctggg ctatgcccgc gtcagcaccg
2881 acgaccagga cttgaccaac caacgggccg aactgcacgc ggccggctgc accaagctgt
2941 tttccgagaa gatcaccggc accaggcgcg accgcccgga gctggccagg atgcttgacc
3001 acctacgccc tggcgacgtt gtgacagtga ccaggctaga ccgcctggcc cgcagcaccc
3061 gcgacctact ggacattgcc gagcgcatcc aggaggccgg cgcgggcctg cgtagcctgg
3121 cagagccgtg ggccgacacc accacgccgg ccggccgcat ggtgttgacc gtgttcgccg
3181 gcattgccga gttcgagcgt tccctaatca tcgaccgcac ccggagcggg cgcgaggccg
3241 ccaaggcccg aggcgtgaag tttggccccc gccctaccct caccccggca cagatcgcgc
3301 acgcccgcga gctgatcgac caggaaggcc gcaccgtgaa agaggcggct gcactgcttg
3361 gcgtgcatcg ctcgaccctg taccgcgcac ttgagcgcag cgaggaagtg acgcccaccg
3421 aggccaggcg gcgcggtgcc ttccgtgagg acgcattgac cgaggccgac gccctggcgg
3481 ccgccgagaa tgaacgccaa gaggaacaag catgaaaccg caccaggacg gccaggacga
3541 accgtttttc attaccgaag agatcgaggc ggagatgatc gcggccgggt acgtgttcga
3601 gccgcccgcg cacgtctcaa ccgtgcggct gcatgaaatc ctggccggtt tgtctgatgc
3661 caagctggcg gcctggccgg ccagcttggc cgctgaagaa accgagcgcc gccgtctaaa
3721 aaggtgatgt gtatttgagt aaaacagctt gcgtcatgcg gtcgctgcgt atatgatgcg
3781 atgagtaaat aaacaaatac gcaaggggaa cgcatgaagg ttatcgctgt acttaaccag
3841 aaaggcgggt caggcaagac gaccatcgca acccatctag cccgcgccct gcaactcgcc
3901 ggggccgatg ttctgttagt cgattccgat ccccagggca gtgcccgcga ttgggcggcc
3961 gtgcgggaag atcaaccgct aaccgttgtc ggcatcgacc gcccgacgat tgaccgcgac
4021 gtgaaggcca tcggccggcg cgacttcgta gtgatcgacg gagcgcccca ggcggcggac
4081 ttggctgtgt ccgcgatcaa ggcagccgac ttcgtgctga ttccggtgca gccaagccct
4141 tacgacatat gggccaccgc cgacctggtg gagctggtta agcagcgcat tgaggtcacg
4201 gatggaaggc tacaagcggc ctttgtcgtg tcgcgggcga tcaaaggcac gcgcatcggc
4261 ggtgaggttg ccgaggcgct ggccgggtac gagctgccca ttcttgagtc ccgtatcacg
4321 cagcgcgtga gctacccagg cactgccgcc gccggcacaa ccgttcttga atcagaaccc
4381 gagggcgacg ctgcccgcga ggtccaggcg ctggccgctg aaattaaatc aaaactcatt
4441 tgagttaatg aggtaaagag aaaatgagca aaagcacaaa cacgctaagt gccggccgtc
4501 cgagcgcacg cagcagcaag gctgcaacgt tggccagcct ggcagacacg ccagccatga
4561 agcgggtcaa ctttcagttg ccggcggagg atcacaccaa gctgaagatg tacgcggtac
4621 gccaaggcaa gaccattacc gagctgctat ctgaatacat cgcgcagcta ccagagtaaa
4681 tgagcaaatg aataaatgag tagatgaatt ttagcggcta aaggaggcgg catggaaaat
4741 caagaacaac caggcaccga cgccgtggaa tgccccatgt gtggaggaac gggcggttgg
4801 ccaggcgtaa gcggctgggt tgtctgccgg ccctgcaatg gcactggaac ccccaagccc
4861 gaggaatcgg cgtgacggtc gcaaaccatc cggcccggta caaatcggcg cggcgctggg
4921 tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac gcatcgaggc
4981 agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca aagaatcccg
5041 gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg acgagcaacc
5101 agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca gcatcatgga
5161 cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga tccgctacga
5221 gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca gtgtgtggga
5281 ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc gataccggga
5341 agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg tactcaagtt
5401 ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct gcattcggtt
5461 aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc gcctggtgac
5521 ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg aaaccgggcg
5581 gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca cagaaggcaa
5641 gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg gcatcggccg
5701 ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat ggttgttcaa
5761 gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt tcaccgtgcg
5821 caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg cggggcaggc
5881 tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat ccgccggttc
5941 ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag gtcgaaaagg
6001 tctctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg ggaaccggaa
6061 cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt aagtgactga
6121 tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta ttaaaactct
6181 taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag agctgcaaaa
6241 agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc ggcctatcgc
6301 ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag ggcgcggaca
6361 agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc ctcgcgcgtt
6421 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
6481 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
6541 gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact ggcttaacta
6601 tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag
6661 atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca ctgactcgct
6721 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
6781 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
6841 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
6901 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
6961 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
7021 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
7081 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
7141 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
7201 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
7261 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
7321 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
7381 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
7441 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
7501 gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtact aaaacaattc
7561 atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca gtaagtcaaa
7621 aaatagctcg acatactgtt cttccccgat atcctccctg atcgaccgga cgcagaaggc
7681 aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc cacttacttt
7741 gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga caagttcctc
7801 ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa atggagtgtc
7861 ttcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt aatccaattc
7921 ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga tggagtgaaa
7981 gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt catcttcata
8041 ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc agccatcatg
8101 ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata gcatcatgtc
8161 cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca tttttaaata
8221 taggttttca ttttctccca ccagcttata taccttagca ggagacattc cttccgtatc
8281 ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc tcattttagc
8341 catttattat ttccttcctc ttttctacag tatttaaaga taccccaaga agctaattat
8401 aacaagacga actccaattc actgttcctt gcattctaaa accttaaata ccagaaaaca
8461 gctttttcaa agttgttttc aaagttggcg tataacatag tatcgacgga gccgattttg
8521 aaaccgcggt gatcacaggc agcaacgctc tgtcatcgtt acaatcaaca tgctaccctc
8581 cgcgagatca tccgtgtttc aaacccggca gcttagttgc cgttcttccg aatagcatcg
8641 gtaacatgag caaagtctgc cgccttacaa cggctctccc gctgacgccg tcccggactg
8701 atgggctgcc tgtatcgagt ggtgattttg tgccgagctg ccggtcgggg agctgttggc
8761 tggctggtgg caggatatat tgtggtgtaa acaaattgac gcttagacaa cttaataaca
8821 cattgcggac gtttttaatg tactgaatta acgccgaatt aattcctagg ccaccatgtt
8881 gggcccggcg cgccaagctt gcatgcctgc aggtcaacat ggtggagcac gacactctcg
8941 tctactccaa gaatatcaaa gatacagtct cagaagacca gagggctatt gagacttttc
9001 aacaaagggt aatatcggga aacctcctcg gattccattg cccagctatc tgtcacttca
9061 tcgaaaggac agtagaaaag gaagatggct tctacaaatg ccatcattgc gataaaggaa
9121 aggctatcgt tcaagatgcc tctaccgaca gtggtcccaa agatggaccc ccacccacga
9181 ggaacatcgt ggaaaaagaa gacgttccaa ccacgtcttc aaagcaagtg gattgatgtg
9241 atggtcaaca tggtggagca cgacactctc gtctactcca agaatatcaa agatacagtc
9301 tcagaagacc agagggctat tgagactttt caacaaaggg taatatcggg aaacctcctc
9361 ggattccatt gcccagctat ctgtcacttc atcgaaagga cagtagaaaa ggaagatggc
9421 ttctacaaat gccatcattg cgataaagga aaggctatcg ttcaagatgc ctctaccgac
9481 agtggtccca aagatggacc cccacccacg aggaacatcg tggaaaaaga agacgttcca
9541 accacgtctt caaagcaagt ggattgatgt gatatctcca ctgacgtaag ggatgacgca
9601 caatcccact atccttcgca agacccttcc tctatataag gaagttcatt tcatttggag
9661 agg
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