pEKEx2 vector (V007586)

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LOCUS       40924_17299        8161 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle-expression vector pEKEx2, complete sequence.
ACCESSION   .
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)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 8161)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (30-MAR-2004) Biotechnology, Forschungszentrum, Leo-Brand, Juelich, 
            NRW 52425, Germany"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8161
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(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
                     /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
                     /label=AmpR promoter
     primer_bind     complement(923..939)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(947..963)
                     /label=lac operator
                     /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    complement(1016..1037)
                     /label=CAP binding site
                     /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..2958
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     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(6724..7752)
                     /codon_start=1
                     /label=lacI
                     /note="lac repressor"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ASA"
     promoter        complement(7753..7830)
                     /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
                     /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)."