pEC-XK99-Gold vector (V015852)

LOCUS       62056_8735        7054 bp DNA     circular SYN 22-JUN-2020
DEFINITION  Shuttle expression vector pEC-XK99-Gold, complete sequence.
ACCESSION   MT521917
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7054)
  AUTHORS   Walter T, Al Medani N, Burgardt A, Cankar K, Ferrer L, Kerbs A, Lee 
            JH, Mindt M, Risse JM, Wendisch VF.
  TITLE     Fermentative N-Methylanthranilate Production by Engineered 
            Corynebacterium glutamicum
  JOURNAL   Microorganisms 8 (6), E866 (2020)
  PUBMED    32521697
REFERENCE   2  (bases 1 to 7054)
  AUTHORS   Cankar K, Walter T.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-MAY-2020) Chair of Genetics of Prokaryotes, Bielefeld 
            University, Universitaetsstrasse 25, Bielefeld, NRW 33615, Germany
REFERENCE   3  (bases 1 to 7054)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Microorganisms"; date: "2020"; volume: "8"; issue: "6"; pages: 
            "E866"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-MAY-2020) Chair of Genetics of Prokaryotes, Bielefeld 
            University, Universitaetsstrasse 25, Bielefeld, NRW 33615, Germany"
FEATURES             Location/Qualifiers
     source          1..7054
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      294..380
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     terminator      472..499
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     gene            complement(738..953)
                     /gene="per"
                     /label=per
     CDS             complement(738..953)
                     /codon_start=1
                     /transl_table=11
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="QKU37006.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(1596..3062)
                     /gene="repA"
                     /label=repA
     CDS             complement(1596..3062)
                     /codon_start=1
                     /transl_table=11
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /protein_id="QKU37007.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERAARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVG
                     ADGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATS
                     NARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELA
                     GELGVSERRIQQVRKALGMEAKRGRPRAEN"
     CDS             3592..4383
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      4486..5074
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(5260..5400)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     promoter        5586..5663
                     /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             5664..6743
                     /label=lacI
                     /note="lac repressor"
     promoter        6978..7007
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"
     protein_bind    7015..7031
                     /label=lac operator
                     /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)."