• 商品货号：BR709

• 规格：10μl质粒
• 启动子：CaMV 35S promoter
• 复制子：pVS1 oriV，ori
• 质粒分类：植物系列，RNAi载体
• 质粒大小：9663bp
• 原核抗性：Kan
• 克隆菌株：DH5α
• 表达宿主：植物细胞
• 5测序引物：35S:GACGCACAATCCCACTATCC
• 3测序引物：根据序列设计引物
• 备注：Tobacco rattle virus RNA2-based VIGS vector
• 质粒简介：     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.
• 质粒图谱1： png/1-1F92Q43646457.png

质粒序列： 点击查看序列

LOCUS       Exported                9663 bp ds-DNA     circular SYN 28-SEP-2017

DEFINITION  synthetic circular DNA

ACCESSION   .

VERSION     .

KEYWORDS    .

SOURCE      synthetic DNA construct

  ORGANISM  synthetic DNA construct

REFERENCE   1  (bases 1 to 9663)

  AUTHORS   .

  TITLE     Direct Submission

  JOURNAL   Exported Sep 28, 2017 from MiaoLingPlasmid

            http://www.fenghbio.cn

FEATURES             Location/Qualifiers

     source          1..9663

                     /organism="synthetic DNA construct"

                     /mol_type="other DNA"

     misc_feature    1..1639

                     /label=TRV strain ppk20 RNA2 5'-sequence

     misc_feature    1647..1707

                     /label=MCS

     misc_feature    1709..2103

                     /label=TRV strain ppk20 RNA2 3'-sequence

     CDS             2164..2181

                     /codon_start=1

                     /product="6xHis affinity tag"

                     /label=6xHis

                     /translation="HHHHHH"

     terminator      2216..2468

                     /label=NOS terminator

                     /note="nopaline synthase terminator and poly(A) signal"

     misc_feature    2490..2514

                     /label=RB T-DNA repeat

                     /note="right border repeat from nopaline C58 T-DNA"

     CDS             3814..4443

                     /codon_start=1

                     /product="stability protein from the Pseudomonas plasmid 

                     pVS1 (Heeb et al., 2000)"

                     /label=pVS1 StaA

                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR

                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP

                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI

                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"

     CDS             4877..5944

                     /codon_start=1

                     /product="replication protein from the Pseudomonas plasmid 

                     pVS1 (Heeb et al., 2000)"

                     /label=pVS1 RepA

                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA

                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR

                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR

                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL

                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA

                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV

                     MRYRNLIEGEASAGS"

     rep_origin      6010..6204

                     /label=pVS1 oriV

                     /note="origin of replication for the Pseudomonas plasmid 

                     pVS1 (Heeb et al., 2000)"

     misc_feature    6548..6688

                     /label=bom

                     /note="basis of mobility region from pBR322"

     rep_origin      complement(6874..7462)

                     /direction=LEFT

                     /label=ori

                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 

                     replication"

     CDS             complement(7549..8343)

                     /codon_start=1

                     /gene="aphA-3"

                     /product="aminoglycoside phosphotransferase"

                     /label=KanR

                     /note="confers resistance to kanamycin"

                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM

                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED

                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT

                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA

                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"

     misc_feature    8768..8792

                     /label=LB T-DNA repeat

                     /note="left border repeat from nopaline C58 T-DNA"

     misc_feature    8895..9663

                     /label=CaMV 35S promoter from pCASS2

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