LOCUS       sequence-100323-        1687 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expression of pHlash in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1687)
  AUTHORS   Zhang Y, Xie Q, Robertson JB, Johnson CH
  TITLE     pHlash: a new genetically encoded and ratiometric luminescence 
            sensor of intracellular pH.
  JOURNAL   PLoS One. 2012;7(8):e43072. doi: 10.1371/journal.pone.0043072. Epub 
            2012 Aug 14.
  PUBMED    22905204
REFERENCE   2  (bases 1 to 1687)
  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: "PLoS One.";
            date: "2012"; volume: "7(8):e43072. doi"; pages: " 
            10.1371/journal.pone.0043072. Epub 2012 Aug 14"
COMMENT     Sequence Label: pcDNA3.1+-pHlash
FEATURES             Location/Qualifiers
     source          1..1687
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             8..940
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGAALAFHYAYEHQDRIKAIVHMESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETVLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             950..1684
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     1087..1108
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1211..1232)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1472..1491)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
ORIGIN
        1 ggatccaatg gcttccaagg tgtacgaccc cgagcaacgc aaacgcatga tcactgggcc
       61 tcagtggtgg gctcgctgca agcaaatgaa cgtgctggac tccttcatca actactatga
      121 ttccgagaag cacgccgaga acgccgtgat ttttctgcat ggtaacgcta cctccagcta
      181 cctgtggagg cacgtcgtgc ctcacatcga gcccgtggct agatgcatca tccctgatct
      241 gatcggaatg ggtaagtccg gcaagagcgg gaatggctca tatcgcctcc tggatcacta
      301 caagtacctc accgcttggt tcgagctgct gaaccttcca aagaaaatca tctttgtggg
      361 ccacgactgg ggggctgctc tggcctttca ctacgcctac gagcaccaag acaggatcaa
      421 ggccatcgtc catatggaga gtgtcgtgga cgtgatcgag tcctgggacg agtggcctga
      481 catcgaggag gatatcgccc tgatcaagag cgaagagggc gagaaaatgg tgcttgagaa
      541 taacttcttc gtcgagaccg tgctcccaag caagatcatg cggaaactgg agcctgagga
      601 gttcgctgcc tacctggagc cattcaagga gaagggcgag gttagacggc ctaccctctc
      661 ctggcctcgc gagatccctc tcgttaaggg aggcaagccc gacgtcgtcc agattgtccg
      721 caactacaac gcctaccttc gggccagcga cgatctgcct aagctgttca tcgagtccga
      781 ccctgggttc ttttccaacg ctattgtcga gggagctaag aagttcccta acaccgagtt
      841 cgtgaaggtg aagggcctcc acttcctcca ggaggacgct ccagatgaaa tgggtaagta
      901 catcaagagc ttcgtggagc gcgtgctgaa gaacgagcag gccgagctca tggacggcgg
      961 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1021 gcccgacaac cactacctga gctaccagtc cgccctgagc aaagacccca acgagaagcg
     1081 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1141 gctgtacaag ggtggcagcg gtggcatggt gagcaagggc gaggagctgt tcaccggggt
     1201 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg
     1261 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagctgatct gcaccaccgg
     1321 caagctgccc gtgccctggc ccaccctcgt gaccaccctg ggctacggcc tgcagtgctt
     1381 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg
     1441 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga
     1501 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa
     1561 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta
     1621 tatcaccgcc gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat
     1681 cgagtaa
//
