LOCUS       sequence-75274-e         693 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Destination plasmid (Gateway cloning) for cumate-inducible 
            expression of 3xFLAG-tagged fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 693)
  AUTHORS   Kaczmarczyk A, Vorholt JA, Francez-Charlot A
  TITLE     Cumate-inducible gene expression system for sphingomonads and other 
            Alphaproteobacteria.
  JOURNAL   Appl Environ Microbiol. 2013 Nov;79(21):6795-802. doi: 
            10.1128/AEM.02296-13. Epub 2013 Aug 30.
  PUBMED    23995928
REFERENCE   2  (bases 1 to 693)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1128/AEM.02296-13"; journalName: "Appl Environ Microbiol"; date:
            "2013-11"; volume: "79"; issue: "21"; pages: "6795-802"
COMMENT     Sequence Label: pQFD
FEATURES             Location/Qualifiers
     source          1..693
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(join(101..107,108..125,126..131))
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(156..280)
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(302..367)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     protein_bind    392..419
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
     protein_bind    462..489
                     /label=CuO
                     /bound_moiety="CymR"
                     /note="CymR-binding P2 operator sequence from the p-cmt 
                     operon of Pseudomonas putida (Mullick et al., 2006)"
ORIGIN
        1 cattgggata tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta
       61 gctcctgaaa atctcgacgg atcctaactc aaaatccaca cattatacga gccggaagca
      121 taaagtgtaa agcctggggt gcctaatgcg gccgccatag tgactggata tgttgtgttt
      181 tacagtatta tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat
      241 cattttacgt ttctcgttca gcttttttgt acaaacttgt ctctagagtc atattaataa
      301 gcttgtcgtc gtcatccttg tagtcgatgt cgtggtcctt atagtcgccg tcgtggtcct
      361 tgtagtccat gcggaagctt cctctactag ttacaaacag accagattgt ctgtttgtta
      421 accggcgcag ttagcggctt cgcggcgatc cgtcaaccat gtacaaacag accagattgt
      481 ctgtttgttc cggtacattg acatatgtat ccgctcatga tacaataacc ctgatgctag
      541 cagggagaga ccccgaatgg tcatcatgtc gccgaagcgc cgcacccagg ccgagcgcgc
      601 gatggaaacc cagggcaagc tgatcgccgc cgccctgggc gtcctgcgcg agaagggcta
      661 tgccggcttc cgcatcgccg acgtcccggg cgc
//
