LOCUS       Exported               11797 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector PC-GW-Kan, complete sequence.
ACCESSION   KP826769
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11797)
  AUTHORS   Dalal J, Yalamanchili R, La Hovary C, Ji M, Rodriguez-Welsh M, 
            Aslett D, Ganapathy S, Grunden A, Sederoff H, Qu R.
  TITLE     A novel gateway-compatible binary vector series (PC-GW) for flexible
            cloning of multiple genes for genetic transformation of plants
  JOURNAL   Plasmid 81, 55-62 (2015)
  PUBMED    26188330
REFERENCE   2  (bases 1 to 11797)
  AUTHORS   Dalal J, Yalamanchili R, Hovary CL, Ji M, Rodriguez-Welsh M, Aslett 
            D, Ganapathy S, Grunden A, Sederoff H, Qu R.
  TITLE     Direct Submission
  JOURNAL   Submitted (19-FEB-2015) Crop Science, North Carolina State 
            University, Campus Box 7287, Raleigh, NC 27695, USA
REFERENCE   3  (bases 1 to 11797)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11797
                     /organism="Binary vector PC-GW-Kan"
                     /mol_type="other DNA"
                     /db_xref="taxon:1642504"
     CDS             1219..1848
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             2282..3349
                     /codon_start=1
                     /product="replication protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 RepA
                     /translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
                     AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
                     DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
                     VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
                     ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
                     RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
                     MRYRNLIEGEASAGS"
     rep_origin      3415..3609
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    3953..4093
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4954..5748)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin resistance protein"
                     /label=aadA
                     /note="AadA"
                     /protein_id="AKC88570.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(4954..5748)
                     /gene="aadA"
                     /label=aadA
     misc_feature    6173..6198
                     /label=left border T-DNA repeat
                     /note="left border T-DNA repeat"
     misc_feature    6173..6197
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     polyA_signal    6275..6449
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(6506..7303)
                     /codon_start=1
                     /gene="NPTII"
                     /product="neomycin phosphotransferase"
                     /label=NPTII
                     /protein_id="AKC88571.1"
                     /translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
                     PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
                     SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
                     QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
                     ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            complement(6506..7303)
                     /gene="NPTII"
                     /label=NPTII
     regulatory      complement(7334..8114)
                     /regulatory_class="promoter"
                     /note="CamV 35S2 promoter"
     promoter        complement(7366..8043)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    8234..8255
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8270..8300
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8308..8324
                     /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     8332..8348
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    8364..11463
                     /label=PC-GW
                     /note="PC-GW"
     regulatory      8478..9313
                     /regulatory_class="promoter"
                     /note="35S promoter from PUGW5 cloning vector"
     promoter        8965..9310
                     /label=CaMV 35S promoter
                     /note="strong constitutive promoter from cauliflower mosaic
                     virus"
     misc_feature    9336..11134
                     /label=Gateway region from pEarleyGate100
                     /note="Gateway region from pEarleyGate100"
     misc_recomb     9336..9518
                     /label=attR1 from pEarleyGate100
                     /note="attR1 from pEarleyGate100"
     protein_bind    9399..9523
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        9548..9578
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             9632..10291
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPVSRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             10633..10938
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="AKC88572.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            10633..10938
                     /gene="ccdB"
                     /label=ccdB
     misc_recomb     11010..11134
                     /label=attR2 from pEarleyGate100
                     /note="attR2 from pEarleyGate100"
     protein_bind    complement(11010..11134)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     regulatory      11238..11463
                     /regulatory_class="terminator"
                     /note="35S terminator from pSIM1 cloning vector"
     primer_bind     complement(11473..11489)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    11692..11716
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    11711..11736
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"