LOCUS       Exported                4569 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  C. glutamicum-E. coli shuttle vector pCRB12, complete sequence.
ACCESSION   HM126494
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4569)
  AUTHORS   Okibe N, Suzuki N, Inui M, Yukawa H.
  TITLE     Antisense-RNA-mediated plasmid copy number control in pCG1-family 
            plasmids, pCGR2 and pCG1, in Corynebacterium glutamicum
  JOURNAL   Microbiology (Reading, Engl.) 156 (PT 12), 3609-3623 (2010)
  PUBMED    20798162
REFERENCE   2  (bases 1 to 4569)
  AUTHORS   Noburyu R, Inui M.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2010) Molecular Microbiology and Biotechnology 
            Group, Research Institute of Innovative Technology for the Earth 
            (RITE), 9-2 Kizugawadai, Kizugawa, Kyoto 619-0292, Japan
REFERENCE   3  (bases 1 to 4569)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4569
                     /organism="C. glutamicum-E. coli shuttle vector pCRB12"
                     /mol_type="other DNA"
                     /db_xref="taxon:931131"
     CDS             306..1763
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="pCG1 RepA"
                     /protein_id="ADR57710.1"
                     /translation="MTLADSRDIDTPAWKFSADLFDTHPELALRSRGWTSEDRREFLAH
                     LGRENFQGSKTRDFASAWIKDPDTGETQPKLYRVGSKSLARCQYVALTHAQHAAVLVLD
                     IDVPSHQAGGKIEHVNPEVYAILERWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAAAG
                     MSSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYRWHAQHNRVDRLADL
                     MEVARMISGTEKPKKRYEQEFSSGRARIEAARKATAEAKALATLEASLPSAAEASGELI
                     DGVRVLWTAPGRAARDETAFRHALTVGYQLKAAGERLKDTKIIDAYERAYTVAQAVGAD
                     GREPDLPPMRDRQTMARRVRGYVAKGQPVVPARQTETQSSRGRKALATMGRRGGKKAAE
                     RWKDPNSEYARAQREKLAKSSQRQARKAKGNRLTIAGWFMTVEGETGSWPTINEAMSEF
                     SVSRQTVNRALKSAGIELPRGRRKASQ"
     gene            306..1763
                     /gene="repA"
                     /label=repA
     regulatory      1912..2011
                     /regulatory_class="promoter"
                     /note="lac promoter"
     protein_bind    1939..1960
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1975..2005
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2013..2029
                     /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     2037..2053
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2049..2357
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="beta-galactosidase alpha peptide"
                     /label=lacZ-alpha
                     /protein_id="ADR57709.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGECDLFNKAAVPSSQRNALPVLQPINQF"
     gene            2049..2357
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     misc_feature    2063..2119
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2123..2139)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2358..3173)
                     /codon_start=1
                     /product="kanamycin resistance protein"
                     /label=kanamycin resistance protein
                     /protein_id="ADR57711.1"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      3553..4141
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"