pYL156 (TRV RNA2) vector (V017513)

LOCUS       Exported               11003 bp DNA     circular SYN 06-DEC-2024
DEFINITION  Modified Tobacco Rattle Virus RNA 2; used for virus induced gene 
            silencing in plants along with pYL192 (TRV RNA1).
ACCESSION   .
VERSION     .
KEYWORDS    pYL156 (TRV RNA2)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11003)
  AUTHORS   Liu Y, Schiff M, Marathe R, Dinesh-Kumar SP
  TITLE     Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for 
            N-mediated resistance to tobacco mosaic virus.
  JOURNAL   Plant J. 2002 May;30(4):415-29. doi: 
            10.1046/j.1365-313x.2002.01297.x.
  PUBMED    12028572
REFERENCE   2  (bases 1 to 11003)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1046/j.1365-313x.2002.01297"; journalName: "Plant J"; date: 
            "2002-05"; volume: "30"; issue: "4"; pages: "415-29"
FEATURES             Location/Qualifiers
     source          1..11003
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     source          770..2408
                     /mol_type="other DNA"
                     /note="Tobacco rattle virus RNA2-based T-DNA vector;
                     synonym: pYL156"
                     /db_xref="taxon:188057"
                     /organism="Tobacco rattle virus RNA2-based VIGS vector
                     pTRV2"
     source          2478..2872
                     /mol_type="other DNA"
                     /note="Tobacco rattle virus RNA2-based T-DNA vector;
                     synonym: pYL156"
                     /db_xref="taxon:188057"
                     /organism="Tobacco rattle virus RNA2-based VIGS vector
                     pTRV2"
     promoter        427..769
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     primer_bind     715..735
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"
     misc_feature    770..2408
                     /label=TRV strain ppk20 RNA2 5'-sequence
                     /note="TRV strain ppk20 RNA2 5'-sequence"
     misc_feature    2478..2872
                     /label=TRV strain ppk20 RNA2 3'-sequence
                     /note="TRV strain ppk20 RNA2 3'-sequence"
     CDS             2933..2950
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      2985..3237
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    3259..3283
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             4584..5213
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5642..6715
                     /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      6781..6975
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid
                     pVS1 (Heeb et al., 2000)"
     primer_bind     complement(7211..7233)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    7319..7459
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     7374..7393
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     complement(7474..7491)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7645..8233)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7725..7744)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             8404..9612
                     /codon_start=1
                     /label=IS4-like element ISVsa5 family transposase
                     /translation="MCELDILHDSLYQFCPELHLKRLNSLTLACHALLDCKTLTLTELG
                     RNLPTKARTKHNIKRIDRLLGNRHLHKERLAVYRWHASFICSGNTMPIVLVDWSDIREQ
                     KRLMVLRASVALHGRSVTLYEKAFPLSEQCSKKAHDQFLADLASILPSNTTPLIVSDAG
                     FKVPWYKSVEKLGWYWLSRVRGKVQYADLGAENWKPISNLHDMSSSHSKTLGYKRLTKS
                     NPISCQILLYKSRSKGRKNQRSTRTHCHHPSPKIYSASAKEPWILATNLPVEIRTPKQL
                     VNIYSKRMQIEETFRDLKSPAYGLGLRHSRTSSSERFDIMLLIALMLQLTCWLAGVHAQ
                     KQGWDKHFQANTVRNRNVLSTVRLGMEVLRHSGYTITREDSLVAATLLTQNLFTHGYVL
                     GKL"
     CDS             complement(9658..10452)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    10877..10901
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"