LOCUS       Exported                8161 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle-expression vector pEKEx2, complete sequence.
ACCESSION   AY585307
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8161)
  AUTHORS   Krumbach K, Eggeling L, Eikmanns B.
  TITLE     Direct Submission
  JOURNAL   Submitted (30-MAR-2004) Biotechnology, Forschungszentrum, Leo-Brand,
            Juelich, NRW 52425, Germany
REFERENCE   2  (bases 1 to 8161)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8161
                     /organism="Shuttle-expression vector pEKEx2"
                     /lab_host="Corynebacterium glutamicum"
                     /mol_type="other DNA"
                     /label=E. coli/C. glutamicum shuttle-expression vector
                     /note="E. coli/C. glutamicum shuttle-expression vector"
                     /db_xref="taxon:272747"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(58..74)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      287..373
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      465..492
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        511..602
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(923..939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    947..963
                     /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)."
     promoter        complement(971..1001)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1016..1037
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(1325..1913)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2146..2961
                     /codon_start=1
                     /gene="aph"
                     /product="kanamycin resistance"
                     /label=aph
                     /protein_id="AAS99551.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2146..2961
                     /gene="aph"
                     /label=aph
     CDS             4577..5860
                     /codon_start=1
                     /product="replicase"
                     /label=replicase
                     /note="orf-1 from Corynebacterium glutamicum pBL1"
                     /protein_id="AAS99552.1"
                     /translation="MNTSKEPQVNEGSKVTRARAWRRQNVMYKITNSKALAGCHRWRRD
                     EAVAVSWSSNGASQFEGLQNSHSRWGSPLAELEVMGERRIELAIATKNHLAAGGALMMF
                     VGTVRHNRSQSFAQVEAGIKTAYSSMVKTSQWKKERARYGVEHTYSDYEVTDSWANGWH
                     LHRNMLLFLDRPLSDDELKAFEDSMFSRWSAGVVKAGMDAPLREHGVKLDQVSTWGGDA
                     AKMATYLAKGMSQELTGSATKTASKGSYTPFQMLDMLADQSDAGEDMDAVLVARWREYE
                     VGSKNLRSSWSRGAKRALGIDYIDADVRREMEEELYKLAGLEAPERVESTRVAVALVKP
                     DDWKLIQSDFAVRQYVLDCVDKAKDVAAAQRVANEVLASLGVDSTPCMIVMDDVDLDAV
                     LPTHGDATKRDLNAAVFAGNEQTILRTH"
     promoter        complement(6581..6609)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     CDS             complement(6655..7629)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /protein_id="AAS99550.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRASAAMPAIMACFSPKRLVAGPVTKA"
     gene            complement(6655..7629)
                     /gene="lacI"
                     /label=lacI
     promoter        complement(7753..7830)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     promoter        8064..8092
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    8100..8116
                     /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)."