LOCUS       Exported               12186 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Binary vector pCLKSCLA25, complete sequence.
ACCESSION   EU327498
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 12186)
  AUTHORS   Ma BG, Duan XY, Niu JX, Ma C, Hao QN, Zhang LX, Zhang HP.
  TITLE     Expression of stilbene synthase gene in transgenic tomato using 
            salicylic acid-inducible Cre/loxP recombination system with 
            self-excision of selectable marker
  JOURNAL   Biotechnol. Lett. 31 (1), 163-169 (2009)
  PUBMED    18792793
REFERENCE   2  (bases 1 to 12186)
  AUTHORS   Ma BG, Duan XY, Niu JX, Zhang LX, Zhang HP.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-DEC-2007) Department of Horticulture, Agriculture 
            College, Shihezi University, Shihezi, Xin Jiang 832003, China
REFERENCE   3  (bases 1 to 12186)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..12186
                     /organism="Binary vector pCLKSCLA25"
                     /mol_type="other DNA"
                     /db_xref="taxon:490119"
     misc_feature    complement(980..1980)
                     /label=STA region from pVS1 plasmid
                     /note="STA region from pVS1 plasmid"
     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      complement(2573..3573)
                     /direction=LEFT
                     /note="pVS1-REP; replication origin from pVS1"
     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"
     misc_feature    complement(3983..4243)
                     /label=bom site from pBR322
                     /note="bom site from pBR322"
     rep_origin      complement(4279..4867)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(4383..4663)
                     /direction=LEFT
                     /label=pBR322 origin of replication
                     /note="pBR322 origin of replication"
     CDS             complement(4954..5748)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside phosphotransferase"
                     /label=aadA
                     /note="kanamycin resistance protein amplified from 
                     pIG121Hm"
                     /protein_id="ABY41258.1"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     gene            complement(4954..5748)
                     /gene="aadA"
                     /label=aadA
     repeat_region   6173..6198
                     /label=left border repeat from C58 T-DNA
                     /note="left border repeat from C58 T-DNA"
     misc_feature    6173..6197
                     /label=LB T-DNA repeat
                     /note="left border repeat from nopaline C58 T-DNA"
     regulatory      6265..6450
                     /regulatory_class="terminator"
                     /note="CaMV 3UTR; includes polyA signal"
     polyA_signal    6275..6449
                     /label=CaMV poly(A) signal
                     /note="cauliflower mosaic virus polyadenylation signal"
     regulatory      complement(6456..7236)
                     /regulatory_class="promoter"
                     /note="CaMV 35S promoter"
     promoter        complement(6488..7165)
                     /label=CaMV 35S promoter (enhanced)
                     /note="cauliflower mosaic virus 35S promoter with a 
                     duplicated enhancer region"
     protein_bind    7356..7377
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        7392..7422
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7430..7446
                     /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     7454..7470
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    7530..7563
                     /label=loxP site
                     /note="loxP site"
     protein_bind    7530..7563
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     regulatory      7597..8922
                     /regulatory_class="promoter"
                     /note="pathogenesis-related protein 1a (PR-1a) promoter"
     CDS             8990..10021
                     /codon_start=1
                     /gene="cre"
                     /product="recombinase"
                     /label=cre
                     /protein_id="ABY41259.1"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRTLDSETGAMVRLLE
                     DGD"
     gene            8990..10021
                     /gene="cre"
                     /label=cre
     regulatory      complement(10326..10581)
                     /regulatory_class="terminator"
                     /note="from TEF"
     gene            complement(10380..11736)
                     /label=kanMX
                     /note="yeast selectable marker conferring kanamycin 
                     resistance (Wach et al., 1994)"
     terminator      10380..10577
                     /label=TEF terminator
                     /note="Ashbya gossypii TEF terminator"
     CDS             complement(10583..11392)
                     /codon_start=1
                     /gene="kanMX"
                     /product="geneticin resistance protein"
                     /label=kanMX
                     /protein_id="ABY41260.1"
                     /translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(10583..11392)
                     /gene="kanMX"
                     /label=kanMX
     regulatory      complement(11393..11790)
                     /regulatory_class="promoter"
                     /note="from TEF"
     promoter        complement(11393..11736)
                     /label=TEF promoter
                     /note="Ashbya gossypii TEF promoter"
     misc_feature    11791..11824
                     /label=loxP site
                     /note="loxP site"
     protein_bind    11791..11824
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     complement(11862..11878)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    12081..12105
                     /label=RB T-DNA repeat
                     /note="right border repeat from nopaline C58 T-DNA"
     repeat_region   12100..12125
                     /label=right border T-DNA repeat
                     /note="right border T-DNA repeat"