LOCUS       sequence-106288-        1561 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Voltage sensing domain of Ciona with the A154D/R217Q/R229I mutations
            fused to super ecliptic pHlorin A227D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1561)
  AUTHORS   Piao HH, Rajakumar D, Kang BE, Kim EH, Baker BJ
  TITLE     Combinatorial mutagenesis of the voltage-sensing domain enables the 
            optical resolution of action potentials firing at 60 Hz by a 
            genetically encoded fluorescent sensor of membrane potential.
  JOURNAL   J Neurosci. 2015 Jan 7;35(1):372-85. doi: 
            10.1523/JNEUROSCI.3008-14.2015.
  PUBMED    25568129
REFERENCE   2  (bases 1 to 1561)
  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: 
            "10.1523/JNEUROSCI.3008-14"; journalName: "J Neurosci"; date: 
            "2015-01-7- 7"; volume: "35"; issue: "1"; pages: "372-85"
COMMENT     Sequence Label: Triple mutant
FEATURES             Location/Qualifiers
     source          1..1561
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     33..52
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        33..51
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      71..80
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             845..1561
                     /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="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNDHQVYIMADKQKNGIKA
                     NFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLFTTSTLSKDPNEKRDHMVLLE
                     FVTADGITHGMDELYK"
     primer_bind     complement(875..903)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1498..1518
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa
       61 gctggctagc gccaccatgg agggattcga cggttcagat tttagtcctc cagctgattt
      121 agttggcgtt gacggtgcag tcatgcggaa cgtcgttgac gtcacgataa atggtgacgt
      181 cactgctccg ccgaaagctg cgccaagaaa aagtgaatcg gtaaagaaag ttcattggaa
      241 tgatgtagac caaggaccga gtgaaaaacc agagacaaga caggaggaac gaatagatat
      301 acccgagatt tcaggtctat ggtggggcga gaatgaacat ggagtggacg atgggagaat
      361 ggagatacct actactggtg taggtcgcgt ccagtttcgg atccgagcag tgattgatca
      421 tctagggatg cgagtctttg gagtcttcct aattttcttg gacatcatcc tcatgatcat
      481 tgatctcagt cttatcgata aaagtgaatc ttcacaatcc ttttatgacg ggatggactt
      541 ggctctttct tgttatttca tgctggattt aggattaagg atatttgcct acgggcccaa
      601 gcttttcttc accaaccctt gggaggtggc tgatggtttg attatcgttg tcacattcgt
      661 cgtcacgata ttttacactg tgttagatga atacggtacc gaaacaggag ccgatggttt
      721 ggggcagttg gttgtgttgg cccgtttgct gcgtgtggtt atcttagcaa gaatatttta
      781 ttcccaccaa caaatgaagg cttcaagcag aagaacaata tcacagaaca aaagaagata
      841 tcgcatgagt aaaggagaag aacttttcac tggagttgtc ccaattcttg ttgaattaga
      901 tggtgatgtt aatgggcaca aattttctgt cagtggagag ggtgaaggtg atgcaacata
      961 cggaaaactt acccttaaat ttatttgcac tactggaaaa ctacctgttc catggccaac
     1021 acttgtcact actttaactt atggtgttca atgcttttca agatacccag atcatatgaa
     1081 acggcatgac tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt
     1141 tttcaaagat gacgggaact acaagacacg tgctgaagtc aagtttgaag gtgataccct
     1201 tgttaataga atcgagttaa aaggtattga ttttaaagaa gatggaaaca ttcttggaca
     1261 caaattggaa tacaactata acgatcacca ggtgtacatc atggcagaca aacaaaagaa
     1321 tggaatcaaa gctaacttca aaattagaca caacattgaa gatggaggcg ttcaactagc
     1381 agaccattat caacaaaata ctccaattgg cgatgggccc gtccttttac cagacaacca
     1441 ttacctgttt acaacttcta ctctttcgaa agatcccaac gaaaagagag accacatggt
     1501 ccttcttgag tttgtaacag ctgatgggat tacacatggc atggatgaac tatacaaatg
     1561 a
//
