LOCUS       sequence-460242-        1080 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of a pH sensitive variant of EGFP inserted in the 
            C-peptide of insulin for pH measurement in insulin secretory 
            granules.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1080)
  AUTHORS   De Lorenzi V, Ghignoli S, Bernardi M, Matteoli G, Ferri G, Storti B,
            Bizzarri R, Cardarelli F
  TITLE     Measuring pH in insulin secretory granules by phasor-based 
            fluorescence lifetime imaging of a genetically encoded sensor.
  JOURNAL   Commun Biol. 2025 Feb 25;8(1):304. doi: 10.1038/s42003-025-07758-w.
  PUBMED    40000885
REFERENCE   2  (bases 1 to 1080)
  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"; journalName: "Commun 
            Biol."; date: "2025-02-25"; volume: "8(1):304. doi"; pages: " 
            10.1038/s42003-025-07758-w"
COMMENT     Sequence Label: pCpep-E1GFP
FEATURES             Location/Qualifiers
     source          1..1080
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      367..376
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(373..375,376..378,379..891)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIE"
     primer_bind     complement(418..439)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(679..698)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1026..1047
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 agcagagctg gtttagtgan ccgtcagatc cgctagcgct accggactca gatctcgagc
       61 tcnngcttcg aattctgcag tcgacccacg cgtccgctac aatcaaaaac catcagcaag
      121 caggaaggtt attgtttcaa catggccctg tggatgcgct tcctgcccct gctggccctg
      181 ctcttcctct gggagtccca ccccacccag gcttttgtca agcagcacct ttgtggttcc
      241 cacctggtgg aggctctcta cctggtgtgt ggggagcgtg gcttcttcta cacacccatg
      301 tcccgccgtg aagtggagga cccacaagtg gcacaactgg agctgggtgg aggcccggga
      361 tccatcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      421 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      481 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      541 ccctggccca ccctcgtgac caccctgagc tacggcgtgc agtgcttcag ccgctacccc
      601 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      661 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
      721 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      781 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
      841 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
      901 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      961 cccgacaacc actacctgag ctaccagtcc gccctgagca aagaccccaa cgagaagcgc
     1021 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
//
