LOCUS       Exported               11277 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCX-DG, complete sequence.
ACCESSION   FJ905222
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11277)
  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 11277)
  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 11277)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..11277
                     /organism="Binary vector pCX-DG"
                     /mol_type="other DNA"
                     /db_xref="taxon:643580"
     regulatory      196..655
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        575..621
                     /label=minimal CaMV 35S promoter
                     /note="minimal 35S promoter from cauliflower mosaic virus"
     regulatory      673..682
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             679..1494
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=green fluorescent protein
                     /note="rsGFP"
                     /protein_id="ACR08780.1"
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTMGYGVLCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYKSGLRSRAQASNSAVDGTAGPGSPILVWGLLKAR"
     misc_feature    1393..1458
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1803..2108
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB"
                     /label=ccdB
                     /protein_id="ACR08781.1"
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     gene            1803..2108
                     /gene="ccdB"
                     /label=ccdB
     terminator      2327..2579
                     /label=NOS terminator
                     /note="nopaline synthase terminator and poly(A) signal"
     regulatory      2330..2582
                     /regulatory_class="terminator"
                     /note="Nos terminator"
     misc_feature    2601..2625
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     misc_feature    2619..2644
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"
     misc_feature    complement(3684..4684)
                     /label=pVS1-STA
                     /note="pVS1-STA"
     CDS             3924..4553
                     /codon_start=1
                     /product="stability protein from the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
                     /label=pVS1 StaA
                     /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
                     DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
                     VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
                     LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
     CDS             4987..6054
                     /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(5277..6277)
                     /direction=LEFT
                     /label=pVS1-REP
                     /note="pVS1-REP"
     rep_origin      6120..6314
                     /label=pVS1 oriV
                     /note="origin of replication for the Pseudomonas plasmid 
                     pVS1 (Heeb et al., 2000)"
     misc_feature    6658..6798
                     /label=bom
                     /note="basis of mobility region from pBR322"
     misc_feature    complement(6687..6947)
                     /label=pBR-BOM
                     /note="pBR-BOM"
     rep_origin      complement(6984..7572)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(7087..7367)
                     /direction=LEFT
                     /label=pBR-ORI
                     /note="pBR-ORI"
     CDS             complement(7659..8453)
                     /codon_start=1
                     /gene="aadA"
                     /product="kanamycin resistance protein"
                     /label=aadA
                     /protein_id="ACR08778.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(7659..8453)
                     /gene="aadA"
                     /label=aadA
     misc_feature    8877..8902
                     /label=left border T-DNA repeat
                     /note="left border T-DNA repeat"
     misc_feature    8878..8902
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     misc_feature    complement(8969..9177)
                     /label=CaMV35S polyA signal
                     /note="CaMV35S polyA signal"
     polyA_signal    8980..9154
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     CDS             complement(9194..10219)
                     /codon_start=1
                     /gene="hptII"
                     /product="hygromycin resistance protein"
                     /label=hptII
                     /note="hygromycin phosphotransferase"
                     /protein_id="ACR08779.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     K"
     gene            complement(9194..10219)
                     /gene="hptII"
                     /label=hptII
     regulatory      complement(10253..11033)
                     /regulatory_class="promoter"
                     /note="CaMV35S promoter"
     promoter        complement(10286..10963)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    11154..11175
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        11190..11220
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    11228..11244
                     /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     11252..11268
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"