LOCUS       Exported               12396 bp ds-DNA    circular SYN 06-5-2013
DEFINITION  Improved binary Agrobacterium vector for plant transformation.
ACCESSION   .
VERSION     .
KEYWORDS    pBINPLUS
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12396)
  AUTHORS   van Engelen FA, Molthoff JW, Conner AJ, Nap JP, Pereira A, Stiekema 
            WJ.
  TITLE     pBINPLUS: an improved plant transformation vector based on pBIN19.
  JOURNAL   Transgenic Res. 1995;4:288-90.
  PUBMED    7655517
REFERENCE   2  (bases 1 to 12396)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     This sequence was reconstructed using the original paper. The 
            orientation of the truncated pBR322 origin is uncertain.
FEATURES             Location/Qualifiers
     source          1..12396
                     /organism="synthetic DNA construct"
                     /lab_host="Plant Cells"
                     /mol_type="other DNA"
     rep_origin      1758..2110
                     /direction=RIGHT
                     /note="pBR322 ori (truncated)"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication (truncated)"
     CDS             2871..3665
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin"
                     /protein_id="
                     "
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     CDS             3964..5112
                     /codon_start=1
                     /product="trans-acting replication protein that binds to 
                     and activates oriV"
                     /note="trfA"
                     /translation="MNRTFDRKAYRQELIDAGFSAEDAETIASRTVMRAPRETFQSVGS
                     MVQQATAKIERDSVQLAPPALPAPSAAVERSRRLEQEAAGLAKSMTIDTRGTMTTKKRK
                     TAGEDLAKQVSEAKQAALLKHTKQQIKEMQLSLFDIAPWPDTMRAMPNDTARSALFTTR
                     NKKIPREALQNKVIFHVNKDVKITYTGVELRADDDELVWQQVLEYAKRTPIGEPITFTF
                     YELCQDLGWSINGRYYTKAEECLSRLQATAMGFTSDRVGHLESVSLLHRFRVLDRGKKT
                     SRCQVLIDEEIVVLFAGDHYTKFIWEKYRKLSPTARRMFDYFSSHREPYPLKLETFRLM
                     CGSDSTRVKKWREQVGEACEELRGSGLVEHAWVNDDLVHCKR"
     misc_feature    6708..6732
                     /note="LB T-DNA repeat"
                     /note="left border repeat from nopaline C58 T-DNA"
     promoter        7173..7352
                     /note="NOS promoter"
                     /note="nopaline synthase promoter"
     CDS             7409..8203
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      8593..8845
                     /note="NOS terminator"
                     /note="nopaline synthase terminator and poly(A) signal"
     promoter        9430..9460
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    9468..9484
                     /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     9492..9508
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             9504..9854
                     /codon_start=1
                     /gene="lacZ (fragment)"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /note="lacZ-alpha"
                     /translation="MTMITPSWRAKLACLQVDSRGSPGTELEFLINNSLAVVLQRRDWE
                     NPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGARSFRFLPFLSRHVRRLSPS
                     SSKSGAPFRVPI"
     misc_feature    9526..9598
                     /note="MCS"
                     /note="multiple cloning site"
     primer_bind     complement(9605..9621)
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    9919..9943
                     /note="RB T-DNA repeat"
                     /note="right border repeat from nopaline C58 T-DNA"
     CDS             10408..11058
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline resistance regulatory protein"
                     /note="TetR"
                     /protein_id="
                     "
                     /translation="MTKLQPNTVIRAALDLLNEVGVDGLTTRKLAERLGVQQPALYWHF
                     RNKRALLDALAEAMLAENHTHSVPRADDDWRSFLIGNARSFRQALLAYRDGARIHAGTR
                     PGAPQMETADAQLRFLCEAGFSAGDAVNALMTISYFTVGAVLEEQAGDSDAGERGGTVE
                     QAPLSPLLRAAIDAFDEAGPDAAFEQGLAVIVDGLAKRRLVVRNVEGPRKGDD"
     oriT            11739..11848
                     /note="incP origin of transfer"
     CDS             11881..12252
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /note="traJ"
                     /protein_id="
                     "
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     rep_origin      12379..694
                     /note="oriV"
                     /note="incP origin of replication"