pQF vector (V003855)

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LOCUS       40924_36193        7535 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Expression vector pQF, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7535)
  AUTHORS   Kaczmarczyk A, Vorholt JA, Francez-Charlot A.
  TITLE     Cumate-inducible gene expression system for sphingomonads and other 
            alphaproteobacteria
  JOURNAL   Appl. Environ. Microbiol. 79 (21), 6795-6802 (2013)
  PUBMED    23995928
REFERENCE   2  (bases 1 to 7535)
  AUTHORS   Kaczmarczyk A.
  TITLE     Direct Submission
  JOURNAL   Submitted (12-AUG-2013) Department of Biology, Institute of 
            Microbiology, ETH Zurich, Wolfgang-Pauli-Str. 10, Zurich 8093, 
            Switzerland
REFERENCE   3  (bases 1 to 7535)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7535)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Appl. 
            Environ. Microbiol."; date: "2013"; volume: "79"; issue: "21"; 
            pages: "6795-6802"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (12-AUG-2013) Department of Biology, Institute of Microbiology, ETH 
            Zurich, Wolfgang-Pauli-Str. 10, Zurich 8093, Switzerland"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7535
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      3..712
                     /label=oriV
                     /note="incP origin of replication"
     rep_origin      1105..1693
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1758..2366)
                     /label=CymR
                     /note="cumate repressor (Mullick et al., 2006)"
     regulatory      complement(2382..2390)
                     /regulatory_class="ribosome_binding_site"
     regulatory      complement(2406..2434)
                     /gene="Pbla-mut1T"
                     /label=modified bla promoter
                     /note="modified bla promoter"
                     /regulatory_class="promoter"
     gene            complement(2406..2434)
                     /gene="Pbla-mut1T"
                     /label=Pbla-mut1T
     regulatory      complement(2406..2411)
                     /gene="Pbla-mut1T"
                     /regulatory_class="minus_10_signal"
     regulatory      complement(2429..2434)
                     /gene="Pbla-mut1T"
                     /regulatory_class="minus_35_signal"
     protein_bind    2443..2470
                     /label=CuO
                     /note="CymR-binding P2 operator sequence from the p-cmt
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      2476..2481
                     /gene="PQ5"
                     /regulatory_class="minus_35_signal"
     regulatory      2499..2505
                     /gene="PQ5"
                     /regulatory_class="minus_10_signal"
     protein_bind    2513..2540
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     regulatory      2547..2554
                     /regulatory_class="ribosome_binding_site"
     CDS             2565..2630
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
     CDS             2727..2750
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     regulatory      2775..2815
                     /label=putative transcriptional terminator T193*
                     /note="putative transcriptional terminator T193*"
                     /regulatory_class="terminator"
     primer_bind     complement(2822..2838)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             complement(3283..3924)
                     /label=TetR
                     /note="tetracycline resistance regulatory protein"
     CDS             4030..5226
                     /label=TcR
                     /note="tetracycline efflux protein"
     CDS             complement(5463..6608)
                     /label=trfA
                     /note="trans-acting replication protein that binds to and 
                     activates oriV"
     CDS             complement(6880..7251)
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSL*AYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     oriT            complement(7284..7393)
                     /direction=LEFT
                     /label=oriT
                     /note="incP origin of transfer"