LOCUS       sequence-101787-        1082 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Fusion of Rluc8 and cpVenus. Expression of pHlash in Escherichia 
            coli for protein purification.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1082)
  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 1082)
  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: pRSETb-pHlash
FEATURES             Location/Qualifiers
     source          1..1082
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             15..37
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             57..74
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             78..110
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(114..122,123..137)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             144..1076
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNATSSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGAALAFHYAYEHQDRIKAIVHMESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETVLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKLFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFLQEDAPDEMGKYIKSFVERVLKNEQ"
ORIGIN
        1 cctctagaaa taattttgtt taactttaag aaggagatat acatatgcgg ggttctcatc
       61 atcatcatca tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt
      121 acgacgatga cgataaggat ccaatggctt ccaaggtgta cgaccccgag caacgcaaac
      181 gcatgatcac tgggcctcag tggtgggctc gctgcaagca aatgaacgtg ctggactcct
      241 tcatcaacta ctatgattcc gagaagcacg ccgagaacgc cgtgattttt ctgcatggta
      301 acgctacctc cagctacctg tggaggcacg tcgtgcctca catcgagccc gtggctagat
      361 gcatcatccc tgatctgatc ggaatgggta agtccggcaa gagcgggaat ggctcatatc
      421 gcctcctgga tcactacaag tacctcaccg cttggttcga gctgctgaac cttccaaaga
      481 aaatcatctt tgtgggccac gactgggggg ctgctctggc ctttcactac gcctacgagc
      541 accaagacag gatcaaggcc atcgtccata tggagagtgt cgtggacgtg atcgagtcct
      601 gggacgagtg gcctgacatc gaggaggata tcgccctgat caagagcgaa gagggcgaga
      661 aaatggtgct tgagaataac ttcttcgtcg agaccgtgct cccaagcaag atcatgcgga
      721 aactggagcc tgaggagttc gctgcctacc tggagccatt caaggagaag ggcgaggtta
      781 gacggcctac cctctcctgg cctcgcgaga tccctctcgt taagggaggc aagcccgacg
      841 tcgtccagat tgtccgcaac tacaacgcct accttcgggc cagcgacgat ctgcctaagc
      901 tgttcatcga gtccgaccct gggttctttt ccaacgctat tgtcgaggga gctaagaagt
      961 tccctaacac cgagttcgtg aaggtgaagg gcctccactt cctccaggag gacgctccag
     1021 atgaaatggg taagtacatc aagagcttcg tggagcgcgt gctgaagaac gagcaggccg
     1081 ag
//
