pTGR5 vector (V002732)

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LOCUS       40924_43163        7520 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle expression vector pTGR5, complete sequence.
ACCESSION   .
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   
REFERENCE   3  (bases 1 to 7520)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7520)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Microb. 
            Cell Fact."; date: "2012"; volume: "11"; issue: "1"; pages: "147"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (29-AUG-2012) Genetic Engineering "
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Assembly Method       :: Seqbuilder v. 7
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..7520
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      24..31
                     /label=from Corynebacterium glutamicum sod gene
                     /note="from Corynebacterium glutamicum sod gene"
                     /regulatory_class="ribosome_binding_site"
     CDS             41..757
                     /label=EGFP
                     /note="enhanced GFP"
     CDS             793..810
                     /label=6xHis
                     /note="6xHis affinity tag"
     terminator      940..1026
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     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..4994
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     rep_origin      5097..5685
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    complement(5965..5986)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     CDS             complement(6002..7081)
                     /label=lacI
                     /note="lac repressor"
     promoter        complement(7082..7159)
                     /label=lacI promoter
     promoter        7482..7510
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and
                     lac UV5 promoters"