LOCUS       Exported               12035 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCXUN-FLAG, complete sequence.
ACCESSION   FJ905217
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12035)
  AUTHORS   Chen S, Songkumarn P, Liu J, Wang GL.
  TITLE     A versatile zero background T-vector system for gene cloning and 
            functional genomics
  JOURNAL   Plant Physiol. 150 (3), 1111-1121 (2009)
  PUBMED    19403729
REFERENCE   2  (bases 1 to 12035)
  AUTHORS   Chen S, Songkumarn P, Liu J, Wang G-L.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-APR-2009) Department of Plant Pathology, The Ohio 
            State University, 201 Kottman Hall, 2021 Coffey Rd, Columbus, OH 
            43210, USA
REFERENCE   3  (bases 1 to 12035)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12035
                     /organism="Binary vector pCXUN-FLAG"
                     /mol_type="other DNA"
                     /db_xref="taxon:643589"
     primer_bind     90..106
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     regulatory      complement(115..367)
                     /regulatory_class="terminator"
                     /note="Nos terminator"
     terminator      115..367
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     CDS             728..1033
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="ACR08765.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            728..1033
                     /gene="ccdB"
                     /label=ccdB
     misc_feature    complement(1089..1115)
                     /label=FLAG-tag
                     /note="FLAG-tag"
     CDS             complement(1089..1112)
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     regulatory      complement(1128..3118)
                     /regulatory_class="promoter"
                     /note="maize ubiquitin-1 promoter"
     promoter        complement(1128..3118)
                     /label=Ubi promoter
                     /note="maize polyubiquitin gene promoter"
     primer_bind     complement(3140..3156)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3359..3383
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    3377..3402
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(4442..5442)
                     /label=pVS1-STA
                     /note="pVS1-STA"
     CDS             4682..5311
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             5745..6812
                     /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      complement(6035..7035)
                     /direction=LEFT
                     /label=pVS1-REP
                     /note="pVS1-REP"
     rep_origin      6878..7072
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    7416..7556
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(7445..7705)
                     /label=pBR-BOM
                     /note="pBR-BOM"
     rep_origin      complement(7742..8330)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(7845..8125)
                     /direction=LEFT
                     /label=pBR-ORI
                     /note="pBR-ORI"
     CDS             complement(8417..9211)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin resistance protein"
                     /label=aadA
                     /protein_id="ACR08763.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(8417..9211)
                     /gene="aadA"
                     /label=aadA
     misc_feature    9635..9660
                     /label=left border T-DNA repeat
                     /note="left border T-DNA repeat"
     misc_feature    9636..9660
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(9727..9935)
                     /label=CaMV35S polyA signal
                     /note="CaMV35S polyA signal"
     polyA_signal    9738..9912
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9952..10977)
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin resistance protein"
                     /label=hptII
                     /note="hygromycin phosphotransferase"
                     /protein_id="ACR08764.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(9952..10977)
                     /gene="hptII"
                     /label=hptII
     regulatory      complement(11011..11791)
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        complement(11044..11721)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11912..11933
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11948..11978
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11986..12002
                     /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     12010..12026
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"