LOCUS       Exported               10417 bp ds-DNA    circular SYN 25-4-2013
DEFINITION  Agrobacterium binary vector for foreign gene expression in 
            dicotyledonous plants.
ACCESSION   .
VERSION     .
KEYWORDS    pRI 101-AN
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10417)
  AUTHORS   TaKaRa
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10417
                     /organism="synthetic DNA construct"
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
     rep_origin      complement(57..645)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    889..5524
                     /note="pRi replicator region"
                     /note="replicator region from Agrobacterium plasmid pRiA 
                     4b"
     misc_feature    5531..5555
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     terminator      5893..6145
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             complement(6419..7213)
                     /codon_start=1
                     /gene="aph(3)-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /note="NeoR/KanR"
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     promoter        complement(7270..7449)
                     /note="NOS promoter"
                     /note="nopaline synthase promoter"
     protein_bind    7584..7605
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7620..7650
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    7658..7674
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     7682..7698
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      7720..7972
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     misc_feature    7983..8084
                     /note="MCS"
                     /note="multiple cloning site"
     5UTR           complement(8085..8142)
                     /note="AtADH 5-UTR"
                     /note="translational enhancer from the 5-UTR of the 
                     Arabidopsis thaliana alcohol dehydrogenase gene (Sugio et 
                     al., 2008"
     promoter        complement(8157..8502)
                     /note="CaMV 35S promoter"
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    9148..9172
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             complement(9256..10050)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /protein_id="
                     "
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"