LOCUS       Exported                4497 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pCRB2 DNA, complete sequence.
ACCESSION   AB526352
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4497)
  AUTHORS   Tsuchida Y, Kimura S, Suzuki N, Inui M, Yukawa H.
  TITLE     Characterization of a 24-kb plasmid pCGR2 newly isolated from 
            Corynebacterium glutamicum
  JOURNAL   Appl. Microbiol. Biotechnol. 87 (5), 1855-1866 (2010)
  PUBMED    20552356
REFERENCE   2  (bases 1 to 4497)
  AUTHORS   Gunji W, Inui M, Suzuki N, Yukawa H.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-OCT-2009) Contact:Nobuaki Suzuki Research Institute of
            Innovative Technology for the Earth, Molecular Microbiology and 
            Biotechnology Group; 9-2 kizugawadai, Kizugawa, kyoto 619-0292, 
            Japan URL :http://www.rite.or.jp/
REFERENCE   3  (bases 1 to 4497)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4497
                     /organism="Shuttle vector pCRB2"
                     /mol_type="other DNA"
                     /note="Corynebacterium glutamicum-Escherichia coli shuttle 
                     vector"
                     /db_xref="taxon:684763"
     misc_feature    6..1818
                     /note="derived from Brevibacterium lactofermentum cryptric 
                     plasmid pBL1"
     CDS             529..1734
                     /codon_start=1
                     /gene="rep"
                     /product="replicase"
                     /label=rep
                     /protein_id="BAI66092.1"
                     /translation="MYKITNSKALAGCRRWRRDEAVAVSWSSNGASQFEGLQNSHSRWG
                     SPLAELEVMGERRIELAIATKNHLAAGGALMMFVGTVRHNRSQSFAQVEAGIKTAYSSM
                     VKTSQWKKERARYGVEHTYSDYEVTDSWANGWHLHRNMLLFLDRPLSDDELKAFEDSMF
                     SRWSAGVVKAGMDAPLREHGVKLDQVPTWGGDAAKMATYLAKGMSQELTGSATKTASKG
                     SYTPFQMLDMLADQSDAGEDMDAVLVARWREYEVGSKNLRSSWSRGAKRALGIDYIDAD
                     VRREMEEELYKLAGLEAPERVESTRVAVALVKPDDWKLIQSDFAVRQYVLDCVDKAKDV
                     AAAQRVANEVLASLGVDSTPCMIVMDDVDLDAVLPTHGDATKRDLNAAVFAGNEQTILR
                     TH"
     gene            529..1734
                     /gene="rep"
                     /label=rep
     misc_feature    1825..4497
                     /note="derived from Escherichia coli cloning vector 
                     pHSG298"
     protein_bind    1866..1887
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1902..1932
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1940..1956
                     /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     1964..1980
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1976..2284
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="lacZ alpha fragment"
                     /label=lacZ-alpha
                     /protein_id="BAI66093.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGECDLFNKAAVPSSQRNALPVLQPINQF"
     gene            1976..2284
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     misc_feature    1990..2046
                     /gene="lacZ-alpha"
                     /label=MCS, multiple cloning site
                     /note="MCS, multiple cloning site"
     primer_bind     complement(2050..2066)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(2285..3100)
                     /codon_start=1
                     /gene="aph"
                     /product="aminoglucoside phosphotransferase"
                     /label=aph
                     /protein_id="BAI66094.1"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            complement(2285..3100)
                     /gene="aph"
                     /label=aph
     rep_origin      3480..4068
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"