LOCUS       Exported                5695 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector pEC-K18mob2, complete sequence.
ACCESSION   AF445080
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  AUTHORS   Tauch A, Kirchner O, Loffler B, Gotker S, Puhler A, Kalinowski J.
  TITLE     Efficient electrotransformation of corynebacterium diphtheriae with 
            a mini-replicon derived from the Corynebacterium glutamicum plasmid 
            pGA1
  JOURNAL   Curr. Microbiol. 45 (5), 362-367 (2002)
  PUBMED    12232668
REFERENCE   2  (bases 1 to 5695)
  AUTHORS   Tauch A.
  TITLE     Direct Submission
  JOURNAL   Submitted (08-NOV-2001) Department of Genetics, University of 
            Bielefeld, Universitaetsstrasse 25, Bielefeld D-33615, Germany
REFERENCE   3  (bases 1 to 5695)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5695
                     /organism="Cloning vector pEC-K18mob2"
                     /lab_host="Corynebacterium glutamicum"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:179875"
     CDS             184..1647
                     /codon_start=1
                     /gene="rep"
                     /product="replication protein"
                     /label=rep
                     /note="Rep"
                     /protein_id="AAL38580.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            184..1647
                     /gene="rep"
                     /label=rep
     CDS             2290..2505
                     /codon_start=1
                     /gene="per"
                     /product="positive effector of replication"
                     /label=per
                     /note="Per"
                     /protein_id="AAL38581.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            2290..2505
                     /gene="per"
                     /label=per
     CDS             3024..3818
                     /codon_start=1
                     /gene="aph(3)-IIa"
                     /product="kanamycin resistance protein"
                     /label=aph(3)-IIa
                     /note="Aph(3)-IIa"
                     /protein_id="AAL38582.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            3024..3818
                     /gene="aph(3)-IIa"
                     /label=aph(3)-IIa
     oriT            4156..4265
                     /note="incP origin of transfer"
     misc_feature    4203..4301
                     /label=RP4 mob region
                     /note="RP4 mob region"
     rep_origin      4477..5065
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      5064..5066
                     /label=oriV of Escherichia coli
                     /note="oriV of Escherichia coli"
     protein_bind    5353..5374
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5389..5419
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5427..5443
                     /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     5451..5467
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             5463..85
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITNSSSVPGDPLESTCRHASLALAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRDKLDPHQSCLFATFNKVVKRSFTKKT
                     R"
     misc_feature    5477..5533
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(5537..5553)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"