LOCUS       sequence-80350-e         979 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  FRET-based, Golgi-targeted kinase activity reporter for PKD.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 979)
  AUTHORS   Kunkel MT, Newton AC
  TITLE     Calcium transduces plasma membrane receptor signals to produce 
            diacylglycerol at Golgi membranes.
  JOURNAL   J Biol Chem. 2010 Jul 23;285(30):22748-52. doi: 
            10.1074/jbc.C110.123133. Epub 2010 Jun 2.
  PUBMED    20519514
REFERENCE   2  (bases 1 to 979)
  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: 
            "10.1074/jbc.C110.123133"; journalName: "J Biol Chem"; date: 
            "2010-07-23- 23"; volume: "285"; issue: "30"; pages: "22748-52"
COMMENT     Sequence Label: Golgi-DKAR
FEATURES             Location/Qualifiers
     source          1..979
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     61..80
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        61..79
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(228..230,231..941)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             join(228..230,231..941)
                     /codon_start=1
                     /product="cyan variant of GFP"
                     /label=CFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             join(228..230,231..941)
                     /codon_start=1
                     /product="enhanced CFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mECFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIKA
                     HFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(270..291)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(531..550)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     878..899
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
       61 taatacgact cactataggg agacccaagc ttatgggcaa cttgaagagc gtggcccagg
      121 agcctgggcc accctgcgga cttggactgg gtctgggact tggattgtgc ggcaagcagg
      181 gcccagccac cccggcaggt ggaggtggtg gaggaggtgg aggtggtgtg agcaagggcg
      241 aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc
      301 acaggttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga
      361 agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga
      421 cctggggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca
      481 agtccgccat gcccgaaggc tacgtccagg agcgtaccat cttcttcaag gacgacggca
      541 actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc
      601 tgaagggcat cgacttcaag gaggacggca acatcctggg acacaagctg gagtacaact
      661 acatcagcca caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggcccact
      721 tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga
      781 acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt
      841 ccaagctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga
      901 ccgccgccgg gatcactcac ggcatggacg agctgtacaa ggctggtacc aagggtaatg
      961 gtaggttttt aactttaaa
//
