LOCUS       Exported               14758 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pBI121, complete sequence.
ACCESSION   AF485783
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 14758)
  AUTHORS   Chen PY, Wang CK, Soong SC, To KY.
  TITLE     Complete sequence of the binary vector pBI121 and its application in
            cloning T-DNA insertion from transgenic plants
  JOURNAL   Mol. Breed. 11, 287-293 (2003)
REFERENCE   2  (bases 1 to 14758)
  AUTHORS   To KY.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-FEB-2002) Institute of BioAgricultural Sciences, 
            Academia Sinica, Taipei 11529, Taiwan
REFERENCE   3  (bases 1 to 14758)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14758
                     /organism="Binary vector pBI121"
                     /mol_type="genomic DNA"
                     /note="constructed using pB221 from Clontech Laboratories 
                     and Bin19 described in GenBank Accession Number U09365"
                     /db_xref="taxon:189807"
     misc_feature    complement(13..796)
                     /label=similar to traF in GenBank Accession Number X54459
                     /note="similar to traF in GenBank Accession Number X54459"
     CDS             complement(145..516)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            549..658
                     /note="incP origin of transfer"
     rep_origin      complement(790..1168)
                     /direction=LEFT
                     /note="ColE1 ori; similar to sequence in GenBank Accession 
                     Number V00268"
     misc_feature    complement(1161..2344)
                     /label=similar to tetA in GenBank Accession Number X75761
                     /note="similar to tetA in GenBank Accession Number X75761"
     CDS             complement(1339..1989)
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /label=TetR
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     misc_feature    complement(2454..2478)
                     /label=T-DNA right border
                     /note="T-DNA right border"
     regulatory      2519..2825
                     /regulatory_class="promoter"
                     /note="NOS"
     promoter        2602..2781
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     promoter        2634..2817
                     /function="NOS promoter"
                     /label=NOS promoter
                     /note="nopaline synthase promoter"
     CDS             2838..3632
                     /codon_start=1
                     /gene="nptII"
                     /product="neomycin phosphotransferase II"
                     /label=nptII
                     /protein_id="AAL92039.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            2838..3632
                     /gene="nptII"
                     /label=nptII
     regulatory      4022..4277
                     /regulatory_class="terminator"
                     /note="NOS"
     terminator      4022..4274
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4859..4889
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4913
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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     4921..4937
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      4974..5808
                     /regulatory_class="promoter"
                     /note="CaMV 35S"
     promoter        5461..5806
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     CDS             5845..7656
                     /codon_start=1
                     /gene="gusA"
                     /product="beta-glucuronidase"
                     /label=gusA
                     /note="GUS"
                     /protein_id="AAL92040.1"
                     /translation="MLRPVETPTREIKKLDGLWAFSLDRENCGIDQRWWESALQESRAI
                     AVPGSFNDQFADADIRNYAGNVWYQREVFIPKGWAGQRIVLRFDAVTHYGKVWVNNQEV
                     MEHQGGYTPFEADVTPYVIAGKSVRITVCVNNELNWQTIPPGMVITDENGKKKQSYFHD
                     FFNYAGIHRSVMLYTTPNTWVDDITVVTHVAQDCNHASVDWQVVANGDVSVELRDADQQ
                     VVATGQGTSGTLQVVNPHLWQPGEGYLYELCVTAKSQTECDIYPLRVGIRSVAVKGEQF
                     LINHKPFYFTGFGRHEDADLRGKGFDNVLMVHDHALMDWIGANSYRTSHYPYAEEMLDW
                     ADEHGIVVIDETAAVGFNLSLGIGFEAGNKPKELYSEEAVNGETQQAHLQAIKELIARD
                     KNHPSVVMWSIANEPDTRPQGAREYFAPLAEATRKLDPTRPITCVNVMFCDAHTDTISD
                     LFDVLCLNRYYGWYVQSGDLETAEKVLEKELLAWQEKLHQPIIITEYGVDTLAGLHSMY
                     TDMWSEEYQCAWLDMYHRVFDRVSAVVGEQVWNFADFATSQGILRVGGNKKGIFTRDRK
                     PKSAAFLLQKRWTGMNFGEKPQQGGKQ"
     gene            5845..7656
                     /gene="gusA"
                     /label=gusA
     regulatory      7727..7979
                     /regulatory_class="terminator"
                     /note="NOS"
     terminator      7727..7979
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     primer_bind     complement(7988..8004)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(8621..8646)
                     /label=T-DNA left border
                     /note="T-DNA left border"
     misc_feature    8622..8646
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(9156..10198)
                     /label=similar to tetA in GenBank Accession Number L13842
                     /note="similar to tetA in GenBank Accession Number L13842"
     misc_feature    complement(10199..11680)
                     /label=similar to trfA in GenBank Accession Number X00713
                     /note="similar to trfA in GenBank Accession Number X00713"
     CDS             complement(10242..11390)
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /label=trfA
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    complement(11681..12673)
                     /note="similar to NPTIII gene in GenBank Accession Number 
                     V01547"
     CDS             complement(11689..12483)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     misc_feature    complement(12674..13443)
                     /note="similar to transposable element IS1 in GenBank 
                     Accession Number X58999"
     mobile_element  12675..13442
                     /mobile_element_type="insertion sequence:IS1"
                     /note="prokaryotic transposable element"
     misc_feature    complement(13444..13794)
                     /note="similarity to NPT III gene in GenBank Accession 
                     Number V01547"
     misc_feature    complement(13795..14066)
                     /label=similar to kilA in GenBank Accession Number M62846
                     /note="similar to kilA in GenBank Accession Number M62846"
     rep_origin      14065..18
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      complement(14141..14758)
                     /direction=LEFT
                     /note="ori V; similar to sequence in GenBank Accession 
                     Number M20134"