LOCUS       Exported                4489 bp ds-DNA     circular SYN 17-DEC-2018
DEFINITION  Shuttle vector pBHK19MECA2, complete sequence.
ACCESSION   AY219863
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4489)
  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 4489)
  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 4489)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4489
                     /organism="Shuttle vector pBHK19MECA2"
                     /mol_type="genomic DNA"
                     /note="Escherichia coli-Corynebacterium glutamicum shuttle 
                     vector"
                     /db_xref="taxon:224632"
     promoter        7..25
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(206..223)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(232..248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             755..1549
                     /codon_start=1
                     /gene="aph(3)-IIa"
                     /product="Aph(3)-IIa"
                     /label=aph(3)-IIa
                     /note="aminoglycoside phosphotransferase"
                     /protein_id="AAO65233.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            755..1549
                     /gene="aph(3)-IIa"
                     /label=aph(3)-IIa
     oriT            1887..1996
                     /note="incP origin of transfer"
     misc_feature    1934..2032
                     /label=RP4-mob region
                     /note="RP4-mob region"
     CDS             2749..4200
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AAO65234.1"
                     /translation="MSLQTHAQNAHTKTPTRVERDRGGLDNTPASASDRDLLRDHLGRD
                     VLHGSVTRDFKKAYRRNADGTNSPRMYRFETDALGRCEYAMLTTKQYAAVLVVDVDQVG
                     TAGGDPADLNPYVRDVVRSLITHSVGPAWVGINPTNGKAQFIWLIDPVYADRNGKSAQM
                     KLLAATTRVLGELLDHDPHFSHRFSRNPFYTGKAPTAYRWYRQHNRVMRLGDLIKQVRD
                     MAGHDQFNPTPRQQFSSGRELINAVKTRREEAQAFKALAQDVDAEIAGGLDQYDPELID
                     GVRVLWIVQGTAARDETAFRHALKTGHRLRQQGQRLTDAAIIDAYEHAYNVAHTHGGAG
                     RDNEMPPMRDRQTMARRVRGYVAQSKSETYSGSNAPGKATSSERKALATMGRRGGQKAA
                     QRWKTDPEGKYAQAQRSKLEKTHRKKKAQGRSTKSRISQMVNDQYFQTGTVPTWAEIGA
                     EVGVSRATVARHVAELKKSGDYPDV"
     gene            2749..4200
                     /gene="repA"
                     /label=repA
     protein_bind    4363..4384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4399..4429
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4437..4453
                     /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     4461..4477
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             4473..442
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSFNTTHYREMLTAVPRPAASHLNACCSPGDVDCLNEALI
                     KGPIGHVRLDLARLEDPWYRCGRTSDRYLPLVRVNVNSLAVVLQRRDWENPGVTQLNRL
                     AAHPPFASWRNSEEARTDRPSQQLRSLNGEWRDKLASRCRKHSGRKGC"