LOCUS       Exported               11329 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  An improved agroinfection-compatible Tobacco mosaic virus 
            (TMV)-based transient expression vector that lacks the TMV CP gene 
            coding sequence and expresses GFP..
ACCESSION   .
VERSION     .
KEYWORDS    pJL-TRBO-G
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11329)
  AUTHORS   Lindbo JA
  TITLE     TRBO: a high-efficiency tobacco mosaic virus RNA-based 
            overexpression vector.
  JOURNAL   Plant Physiol. 2007 Dec;145(4):1232-40. Epub 2007 Aug 24.
  PUBMED    17720752
REFERENCE   2  (bases 1 to 11329)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Plant 
            Physiol."; date: "2007-12"; volume: "145(4)"; pages: "1232-40. Epub 
            2007 Aug 24"
FEATURES             Location/Qualifiers
     source          1..11329
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..46
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     misc_feature    58..113
                     /label=TMV Omega
                     /note="translational enhancer from the tobacco mosaic virus
                     5-leader sequence (Gallie et al., 1988)"
     CDS             5811..6530
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(5844..5872)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6467..6487
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     misc_feature    7137..7161
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(7316..8464)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MAQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     CDS             complement(8763..9557)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHLERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      9766..10297
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      9774..10388
                     /label=oriV
                     /note="origin of replication for the bacterial F plasmid"
     misc_feature    10481..10505
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     primer_bind     10585..10601
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     10586..10602
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(11321..11329,1..12)
                     /label=35S promoter
                     /note="CaMV 35S promoter, forward primer"