LOCUS       sequence-315842-         822 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  ratiometric pHluorin (mitchondrial).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 822)
  AUTHORS   Werley CA, Boccardo S, Rigamonti A, Hansson EM, Cohen AE
  TITLE     Multiplexed Optical Sensors in Arrayed Islands of Cells for 
            multimodal recordings of cellular physiology.
  JOURNAL   Nat Commun. 2020 Aug 4;11(1):3881. doi: 10.1038/s41467-020-17607-5.
  PUBMED    32753572
REFERENCE   2  (bases 1 to 822)
  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 
            Commun."; date: "2020-08-4"; volume: "11(1):3881. doi"; pages: " 
            10.1038/s41467-020-17607-5"
COMMENT     Sequence Label: pMOS018: ratiometric pHluorin (mitchondrial)
FEATURES             Location/Qualifiers
     source          1..822
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..75
                     /codon_start=1
                     /product="mitochondrial presequence of human cytochrome c 
                     oxidase subunit VIII (Rizutto et al., 1989)"
                     /label=COX8 presequence
                     /translation="MSVLTPLLLRGLTGSARRLPVPRAK"
     CDS             109..822
                     /codon_start=1
                     /product="pH-sensitive mutant of green fluorescent protein 
                     (Miesenbock et al., 1998)"
                     /label=ratiometric pHluorin
                     /note="responds to a drop in pH with a reversible decrease 
                     in excitation at 395 nm and a corresponding increase in 
                     excitation at 475 nm"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKDDGNILGHKLEYNYNEHLVYIMADKQKNGTKAI
                     FQVHHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLHTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     primer_bind     complement(136..164)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     759..779
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 atgtccgtcc tgacgccgct gctgctgcgg ggcttgacag gctcggcccg gcggctccca
       61 gtgccgcgcg ccaagatcca ttcgttgggt gacccgccag tcgctagcag taaaggagaa
      121 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
      181 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
      241 tttatttgca ctactggaaa actacctgtt ccatggccaa cacttgtcac tactttctct
      301 tatggtgttc aatgcttttc aagataccca gatcatatga aacggcatga ctttttcaag
      361 agtgccatgc ccgaaggtta tgtacaggaa agaactatat ttttcaaaga tgacgggaac
      421 tacaagacac gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
      481 aaaggtattg attttaaaga tgatggaaac attcttggac acaaattgga atacaactat
      541 aacgagcact tggtgtacat catggcagac aaacaaaaga atggtaccaa agctatcttt
      601 caagttcacc acaacattga agatggaggc gttcaactag cagaccatta tcaacaaaat
      661 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgca cacacaatct
      721 gccctttcga aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca
      781 gctgctggga ttacacatgg catggatgaa ctatacaaat ga
//
