LOCUS       Exported                7520 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle expression vector pTGR5, complete sequence.
ACCESSION   JX559328
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7520)
  AUTHORS   Ravasi P, Peiru S, Gramajo H, Menzella HG.
  TITLE     Design and testing of a synthetic biology framework for genetic 
            engineering of Corynebacterium glutamicum
  JOURNAL   Microb. Cell Fact. 11 (1), 147 (2012)
  PUBMED    23134565
REFERENCE   2  (bases 1 to 7520)
  AUTHORS   Ravasi P, Peiru S, Gramajo H, Menzella HG.
  TITLE     Direct Submission
  JOURNAL   Submitted (29-AUG-2012) Genetic Engineering & Fermentation 
            Technology, IBR, Suipacha 590, Rosario 2000, Argentina
REFERENCE   3  (bases 1 to 7520)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Seqbuilder v. 7
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7520
                     /organism="Shuttle expression vector pTGR5"
                     /mol_type="other DNA"
                     /db_xref="taxon:1229160"
     regulatory      18..7289
                     /regulatory_class="promoter"
                     /note="from tac"
     regulatory      24..31
                     /regulatory_class="ribosome_binding_site"
                     /note="from Corynebacterium glutamicum sod gene"
     CDS             41..760
                     /codon_start=1
                     /gene="egfp"
                     /product="enhanced green fluorescent protein"
                     /label=egfp
                     /protein_id="AFQ97506.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            41..760
                     /gene="egfp"
                     /label=egfp
     CDS             793..810
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      940..1026
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     regulatory      943..986
                     /regulatory_class="terminator"
                     /note="rrnbT1"
     regulatory      1118..1147
                     /regulatory_class="terminator"
                     /note="rrnBT2"
     terminator      1119..1146
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     CDS             complement(1352..1567)
                     /codon_start=1
                     /gene="per"
                     /product="Per"
                     /label=per
                     /note="positive effector of replication"
                     /protein_id="AFQ97508.1"
                     /translation="MDLSDVALESDALDAAVDLKTVIGFFRALDTTDAPASRDWASAAS
                     DLETLVADLEELADELRARQRQEDAQ"
     gene            complement(1352..1567)
                     /gene="per"
                     /label=per
     CDS             complement(2210..3673)
                     /codon_start=1
                     /gene="repA"
                     /product="RepA"
                     /label=repA
                     /note="replication protein"
                     /protein_id="AFQ97507.1"
                     /translation="MTLADPQDTVTASAWKLSADLFDTHPEAMRCGSRGWTAEDRRELL
                     AHLGRESFQGSKTRDFASAWIKNPDTGETQPKLYRAGSKALTRCQYVALTHAQHAAVIV
                     LDIDVPSHQAGGKIEHVNPQVYAILEKWARLEKAPAWIGVNPLSGKCQLIWLIDPVYAA
                     AGKTSPNMRLLAATTEEMTRVFGADQAFSHRLSRWPLHVSDDPTAYKWHCQHDRVDRLA
                     DLMEIARTMTGSQKPKKYIEQDFSSGRARIEAAQRATAEAKALAILDASLPSALDASGD
                     LIDGVRVLWTNPERARDETAFRHALTVGYQLKAAGERLKDAKIIDAYEVAYNVAQAVGA
                     DGREPDLPAMRDRLTMARRVRGYVAKGQPVVPARRVETQSSRGRKALATMGRRGAATSN
                     ARRWADPESKYAQETRQRLAEANKRREMTGELLELRVKTAILDARSQSVADPSTRELAG
                     ELGVSERRIQQVRKALGMEAKRGRPRAEN"
     gene            complement(2210..3673)
                     /gene="repA"
                     /label=repA
     CDS             4203..4997
                     /codon_start=1
                     /gene="Aph(3)-IIa"
                     /product="aminoglycoside phosphotransferase"
                     /label=Aph(3)-IIa
                     /protein_id="AFQ97505.1"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     gene            4203..4997
                     /gene="Aph(3)-IIa"
                     /label=Aph(3)-IIa
     rep_origin      5097..5685
                     /label=from Escherichia coli
                     /note="from Escherichia coli"
     CDS             complement(5999..7081)
                     /codon_start=1
                     /gene="lacIq"
                     /product="lac repressor"
                     /label=lacIq
                     /protein_id="AFQ97509.1"
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     gene            complement(5999..7081)
                     /gene="lacIq"
                     /label=lacIq
     promoter        complement(7082..7159)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        7482..7510
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    7518..14
                     /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)."