LOCUS       sequence-127951-        1047 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of lyso-pHluorin in mammalian cells for optical readout 
            of lysosomal acidification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1047)
  AUTHORS   Rost BR, Schneider F, Grauel MK, Wozny C, G Bentz C, Blessing A, 
            Rosenmund T, Jentsch TJ, Schmitz D, Hegemann P, Rosenmund C
  TITLE     Optogenetic acidification of synaptic vesicles and lysosomes.
  JOURNAL   Nat Neurosci. 2015 Nov 9. doi: 10.1038/nn.4161.
  PUBMED    26551543
REFERENCE   2  (bases 1 to 1047)
  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: "Nat 
            Neurosci. 2015 Nov 9. doi: 10.1038/nn.4161."
COMMENT     Sequence Label: pCMV-lyso-pHluorin
FEATURES             Location/Qualifiers
     source          1..1047
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      186..195
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             192..908
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Ng et al., 2002)"
                     /label=superecliptic pHluorin
                     /note="excited strongly by blue light at neutral pH but 
                     minimally fluorescent at acidic pH"
                     /translation="MGSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(225..253)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     848..868
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 ctatataagc agagctggtt tagtgaaccg tcagatccgc tagcaccatg gcggtggaag
       61 gaggaatgaa gtgtgtcaag tttttgctct acgttctcct gctggccttc tgcgcctgtg
      121 cagtgggatt gatcgccatt ggtgtagcgg ttcaggttgt cttgaagcag gccgaagcgg
      181 atctagccac catgggaagt aaaggagaag aacttttcac tggagttgtc ccaattcttg
      241 ttgaattaga tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg
      301 atgcaacata cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc
      361 cttggccaac acttgtcact actttaactt atggtgttca atgcttttca agatacccag
      421 atcatatgaa acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa
      481 gaactatatt tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag
      541 gtgataccct tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca
      601 ttcttggaca caaattggaa tacaactata acgatcacca ggtgtacatc atggcagaca
      661 aacaaaagaa tggaatcaaa gctaacttca aaattagaca caacattgaa gatggaggcg
      721 ttcaactagc agaccattat caacaaaata ctccaattgg cgatgggccc gtccttttac
      781 cagacaacca ttacctgttt acaacttcta ctctttcgaa agatcccaac gaaaagagag
      841 accacatggt ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac
      901 tatacaaaac cggtggagct ccaattaccc atgagactac tgctggctcg ctgttgcctg
      961 tggtcatcat tgcagtgggt gccttcctct tcctggtggc ctttgtgggc tgctgtgggg
     1021 cctgcaagga gaactactgt ctcatga
//
