LOCUS       Exported                3999 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pCRB26 DNA, complete sequence.
ACCESSION   AB526353
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3999)
  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 3999)
  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 3999)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..3999
                     /organism="Shuttle vector pCRB26"
                     /mol_type="other DNA"
                     /note="Corynebacterium glutamicum-Escherichia coli shuttle 
                     vector"
                     /db_xref="taxon:684764"
     misc_feature    1..1362
                     /note="derived from Escherichia coli cloning vector 
                     pHSG398"
     protein_bind    202..223
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        238..268
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    276..292
                     /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     300..316
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             312..542
                     /codon_start=1
                     /gene="lacZ-alpha"
                     /product="lacZ alpha fragment"
                     /label=lacZ-alpha
                     /protein_id="BAI66095.1"
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGE"
     gene            312..542
                     /gene="lacZ-alpha"
                     /label=lacZ-alpha
     misc_feature    326..382
                     /gene="lacZ-alpha"
                     /label=MCS, multiple cloning site
                     /note="MCS, multiple cloning site"
     primer_bind     complement(386..402)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      complement(728..1316)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    1365..2815
                     /note="derived from Corynebacteriumt glutamicum cryptric 
                     plasmid pCASE1"
     CDS             complement(1421..1888)
                     /codon_start=1
                     /gene="rep2"
                     /product="replicase"
                     /label=rep2
                     /protein_id="BAI66096.1"
                     /translation="MNHYLWPKLTVLPGQSINGSSPSHACGQTAPPSTTPHSPQCNPHA
                     GRKAGNEALRCVVRLDILFGGTLAKVYARYPRNGKTARELAERAGMSVRTAQRWTSEPR
                     EVFIKRANEKRARVQELRAKGLSMRAIAAEIGCSVGTVHRYVKEVEEKKTA"
     gene            complement(1421..1888)
                     /gene="rep2"
                     /label=rep2
     CDS             complement(1680..2615)
                     /codon_start=1
                     /gene="rep1"
                     /product="replicase"
                     /label=rep1
                     /protein_id="BAI66097.1"
                     /translation="MKTDRARSCEKVSYMRPTTRPEGRFEQLWLPLWPLASDDLREGIY
                     RTSRKNALDKRYVEANPDALSNLLVVDIDQEDALLRSLWDREDWRPNAVVENPLNGHAH
                     AVWALAEPFTRTEYAKRKPLAYAAAVTEGLRRSVDGDSGYSGLITKNPEHTAWDSHWIT
                     DKLYTLDELRFWLEETGFMPPASWRKTRRFSPVGLGRNCALFESARTWAYREVRKHFGD
                     ADGLGRAIQTTAQALNQELFDEPLPVAEVDCIARSIHKWIITKSRMWTDGAAVYDATFT
                     AMQSARGKKGWQRSAEVRREAGHTLWRNIG"
     gene            complement(1680..2615)
                     /gene="rep1"
                     /label=rep1
     CDS             complement(2849..3874)
                     /codon_start=1
                     /gene="hpt"
                     /product="hygromycin phosphotransferase"
                     /label=hpt
                     /protein_id="BAI66098.1"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     gene            complement(2849..3874)
                     /gene="hpt"
                     /label=hpt
     protein_bind    3923..3939
                     /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(3947..3977)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"