LOCUS       Exported                9967 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pOGOduet, complete sequence.
ACCESSION   MG582594
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9967)
  AUTHORS   Goldbeck O, Seibold GM.
  TITLE     Construction of pOGOduet - An inducible, bicistronic vector for 
            synthesis of recombinant proteins in Corynebacterium glutamicum
  JOURNAL   Plasmid 95, 11-15 (2018)
  PUBMED    29331350
REFERENCE   2  (bases 1 to 9967)
  AUTHORS   Goldbeck O, Seibold GM.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-NOV-2017) Institute of Microbiology and Biotechnology,
            Ulm University, Albert-Einstein-Allee 11, Ulm, BW 89081, Germany
REFERENCE   3  (bases 1 to 9967)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9967
                     /organism="Cloning vector pOGOduet"
                     /mol_type="other DNA"
                     /db_xref="taxon:2073060"
     rep_origin      803..9487
                     /label=OriCg
                     /note="OriCg"
     CDS             complement(1647..2273)
                     /codon_start=1
                     /gene="tetR"
                     /product="TetR"
                     /label=tetR
                     /note="tet repressor"
                     /protein_id="AUR53410.1"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     gene            complement(1647..2273)
                     /gene="tetR"
                     /label=tetR
     CDS             complement(1647..2270)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     regulatory      2570..2869
                     /regulatory_class="promoter"
                     /note="Ptet-tetO"
     protein_bind    2805..2823
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     protein_bind    2839..2855
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     primer_bind     complement(2940..2956)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      3169..3255
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3347..3374
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3393..3484
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3805..3821)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3829..3845
                     /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(3853..3883)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3898..3919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             complement(3971..4945)
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacI"
                     /label=lacIq
                     /note="lac repressor"
                     /protein_id="AUR53409.1"
                     /translation="MAELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSR
                     ADQLGASVVVSMVERSGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPA
                     LFLDVSDQTPINSIIFSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKY
                     LTRNQIQPIAEREGDWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLR
                     VGADISVVGYDDTEDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVS
                     LVKRKTTLAPNTQTASPRASAAMPAIMACFSPKRLVAGPVTKA"
     gene            complement(3971..4945)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(5069..5146)
                     /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."
     regulatory      5235..5448
                     /regulatory_class="promoter"
                     /note="Ptac"
     promoter        5380..5408
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    5416..5432
                     /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     complement(5536..5552)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      5765..5851
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      5943..5970
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     rep_origin      6229..6848
                     /label=OriEc
                     /note="OriEc"
     rep_origin      complement(6235..6823)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             7056..7871
                     /codon_start=1
                     /gene="aph"
                     /product="Aph"
                     /label=aph
                     /protein_id="AUR53408.1"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            7056..7871
                     /gene="aph"
                     /label=aph