pPBEx2 vector (V015722)

LOCUS       62056_18365        7935 bp DNA     circular SYN 21-SEP-2020
DEFINITION  Shuttle-expression vector pPBEx2, complete sequence.
ACCESSION   MT670349
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7935)
  AUTHORS   Bakkes PJ, Ramp P, Bida A, Dohmen-Olma D, Bott M, Freudl R.
  TITLE     Improved pEKEx2-derived expression vectors for tightly controlled 
            production of recombinant proteins in Corynebacterium glutamicum
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7935)
  AUTHORS   Bakkes PJ, Ramp P, Bida A, Dohmen-Olma D, Bott M, Freudl R.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUN-2020) Institute of Bio- and Geosciences; IBG-1: 
            Biotechnology, Forschungszentrum Juelich GmbH, Wilhelm-Johnen 
            Strasse, Juelich, NRW 52425, Germany
REFERENCE   3  (bases 1 to 7935)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUN-2020) Institute of Bio- and Geosciences"; volume: " IBG-1"; 
            pages: " Biotechnology, Forschungszentrum Juelich GmbH, 
            Wilhelm-Johnen Strasse, Juelich, NRW 52425, German"
FEATURES             Location/Qualifiers
     source          1..7935
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..56)
                     /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
     rep_origin      complement(1114..1702)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             1935..2747
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
     CDS             4366..5649
                     /codon_start=1
                     /transl_table=11
                     /product="replicase"
                     /label=replicase
                     /note="Orf-1 from Corynebacterium glutamicum pBL1"
                     /protein_id="QNT09622.1"
                     /translation="MNTSKEPQVNEGSKVTRARAWRRQNVMYKITNSKALAGCHRWRRD
                     EAVAVSWSSNGASQFEGLQNSHSRWGSPLAELEVMGERRIELAIATKNHLAAGGALMMF
                     VGTVRHNRSQSFAQVEAGIKTAYSSMVKTSQWKKERARYGVEHTYSDYEVTDSWANGWH
                     LHRNMLLFLDRPLSDDELKAFEDSMFSRWSAGVVKAGMDAPLREHGVKLDQVSTWGGDA
                     AKMATYLAKGMSQELTGSATKTASKGSYTPFQMLDMLADQSDAGEDMDAVLVARWREYE
                     VGSKNLRSSWSRGAKRALGIDYIDADVRREMEEELYKLAGLEAPERVESTRVAVALVKP
                     DDWKLIQSDFAVRQYVLDCVDKAKDVAAAQRVANEVLASLGVDSTPCMIVMDDVDLDAV
                     LPTHGDATKRDLNAAVFAGNEQTILRTH"
     promoter        complement(6370..6398)
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein
                     gene"
     CDS             complement(6447..7526)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7527..7604)
                     /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        7838..7866
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7874..7890
                     /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)."