LOCUS       Exported                5536 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  C.glutamicum-E.coli shuttle vector pCRB12i, complete sequence.
ACCESSION   KP340794
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5536)
  AUTHORS   Oide S, Gunji W, Moteki Y, Yamamoto S, Suda M, Jojima T, Yukawa H, 
            Inui M.
  TITLE     Thermal and Solvent Stress Cross-Tolerance Conferred to 
            Corynebacterium glutamicum by Adaptive Laboratory Evolution
  JOURNAL   Appl. Environ. Microbiol. 81 (7), 2284-2298 (2015)
  PUBMED    25595768
REFERENCE   2  (bases 1 to 5536)
  AUTHORS   Gunji W, Oide S, Inui M.
  TITLE     Direct Submission
  JOURNAL   Submitted (05-JAN-2015) Microbiology Group, Research Institute of 
            Innovative Technology for the Earth, 9-2 Kizugawadai, Kyoto 
            619-0292, Japan
REFERENCE   3  (bases 1 to 5536)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..5536
                     /organism="C.glutamicum-E.coli shuttle vector pCRB12i"
                     /mol_type="other DNA"
                     /db_xref="taxon:1609093"
     CDS             62..1153
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacIQ"
                     /label=lacIq
                     /note="repressor"
                     /protein_id="AJI43667.1"
                     /translation="MVNVKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAM
                     AELNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVER
                     SGVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGD
                     WSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTED
                     SSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTA
                     SPRALADSLMQLARQVSRLESGQ"
     gene            62..1153
                     /gene="lacIq"
                     /label=lacIq
                     /note="derived from pDW363"
     CDS             71..1153
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     rep_origin      1621..2209
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2757..3572
                     /codon_start=1
                     /gene="npt"
                     /product="neomycin phosphotransferase"
                     /label=npt
                     /note="NPT"
                     /protein_id="AJI43668.1"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEGYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     gene            2757..3572
                     /gene="npt"
                     /label=npt
     misc_feature    3635..5528
                     /note="derived from Corynebacterium glutamicum plasmid 
                     pCG1"