LOCUS       Exported                7018 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle expression vector pEC-XK99E, complete sequence.
ACCESSION   AY219683
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Tools for genetic engineering in the amino acid-producing bacterium 
            Corynebacterium glutamicum
  JOURNAL   J. Biotechnol. 104 (1-3), 287-299 (2003)
  PUBMED    12948646
REFERENCE   2  (bases 1 to 7018)
  AUTHORS   Kirchner O, Tauch A.
  TITLE     Direct Submission
  JOURNAL   Submitted (15-JAN-2003) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 7018)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7018
                     /organism="Shuttle expression vector pEC-XK99E"
                     /mol_type="genomic DNA"
                     /note="Escherichia coli-Corynebacterium glutamicum shuttle 
                     vector"
                     /db_xref="taxon:224638"
     misc_feature    1..57
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     terminator      260..346
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      263..306
                     /regulatory_class="terminator"
                     /note="terminator T1"
     regulatory      438..465
                     /regulatory_class="terminator"
                     /note="terminator T2"
     terminator      438..465
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(705..920)
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AAO65195.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(705..920)
                     /gene="per"
                     /label=per
     CDS             complement(1563..3026)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65196.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            complement(1563..3026)
                     /gene="repA"
                     /label=repA
     CDS             3556..4350
                     /codon_start=1
                     /gene="aph(3)-IIa"
                     /product="Aph(3)-IIa"
                     /label=aph(3)-IIa
                     /note="aminoglycoside phosphotransferase"
                     /protein_id="AAO65197.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3556..4350
                     /gene="aph(3)-IIa"
                     /label=aph(3)-IIa
     rep_origin      4450..5038
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5038
                     /label=oriV
                     /note="oriV"
     misc_feature    5224..5364
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        5550..5627
                     /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."
     CDS             5628..6710
                     /codon_start=1
                     /gene="lacIq"
                     /product="LacIq"
                     /label=lacIq
                     /note="Lac-repressor"
                     /protein_id="AAO65198.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            5628..6710
                     /gene="lacIq"
                     /label=lacIq
     regulatory      6767..7012
                     /regulatory_class="promoter"
                     /note="Ptrc"
     promoter        6942..6971
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    6979..6995
                     /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)."