LOCUS       sequence-76540-e         992 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cumate-inducible expression of ECFP-tagged fusion protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 992)
  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 992)
  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: pQC
FEATURES             Location/Qualifiers
     source          1..992
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(72..137)
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             complement(join(189..899,900..902,903..905))
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(231..252)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     580..599
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     839..860
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     protein_bind    927..954
                     /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 gaattcaaaa aaaaggccgg gggagcgatc ccctggcctt tttttttgcg ggtcacttca
       61 ccggatcctc acttgtcgtc gtcatccttg tagtcgatgt cgtggtcctt atagtcgccg
      121 tcgtggtcct tgtagtccgg atcccaattg gagctcggta cctcgagatc ttctagagtc
      181 atattaatct tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc
      241 agcaggacca tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctgggtgctc
      301 aggtagtggt tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag
      361 tggtcggcga gctgcacgct gccgtcctcg atgttgtggc ggatcttgaa gttggccttg
      421 atgccgttct tctgcttgtc ggcggtgata tagacgttgt ggctgatgta gttgtactcc
      481 agcttgtgcc ccaggatgtt gccgtcctcc ttgaagtcga tgcccttcag ctcgatgcgg
      541 ttcaccaggg tgtcgccctc gaacttcacc tcggcgcggg tcttgtagtt gccgtcgtcc
      601 ttgaagaaga tggtgcgctc ctggacgtag ccttcgggca tggcggactt gaagaagtcg
      661 tgctgcttca tgtggtcggg gtagcggctg aagcactgca cgccccaggt cagggtggtc
      721 acgagggtgg gccagggcac gggcagcttg ccggtggtgc agatgaactt cagggtcagc
      781 ttgccgtagg tggcatcgcc ctcgccctcg ccggacacgc tgaacttgtg gccgtttacg
      841 tcgccgtcca gctcgaccag gatgggcacc accccggtga acagctcctc gcccttgctc
      901 accatgcgga agcttcctct actagttaca aacagaccag attgtctgtt tgttaaccgg
      961 cgcagttagc ggcttcgcgg cgatccgtca ac
//
