LOCUS       sequence-101523-         976 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Red fluorescent protein based pH indicator in bacterial expression 
            vector.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 976)
  AUTHORS   Shen Y, Rosendale M, Campbell RE, Perrais D
  TITLE     pHuji, a pH-sensitive red fluorescent protein for imaging of exo- 
            and endocytosis.
  JOURNAL   J Cell Biol. 2014 Nov 10;207(3):419-32. doi: 10.1083/jcb.201404107.
  PUBMED    25385186
REFERENCE   2  (bases 1 to 976)
  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: "10.1083/jcb"; 
            journalName: "J Cell Biol"; date: "2014-11-10- 10"; volume: "207"; 
            issue: "3"; pages: "419-32"
COMMENT     Sequence Label: pBAD-pHuji
FEATURES             Location/Qualifiers
     source          1..976
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(6..716)
                     /codon_start=1
                     /product="pH-sensitive red fluorescent protein derived from
                     mApple, for imaging of exocytosis and endocytosis (Shen et 
                     al., 2014)"
                     /label=pHuji
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA
                     FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF
                     EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK
                     SEIKYRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(284..303)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             complement(join(732..746,747..755))
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             complement(759..791)
                     /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             complement(795..812)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(916..935)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 agcttttact tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg
       61 tactgttcca cgatggtgta gtcctcgttg tgggacacga tgtccaactt gatgtccacg
      121 atgtaggcgc cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcggcg
      181 gcgtagtggc cgccgtcctt cagcttcagc ctatacttaa tctcgctctt cagggcgccg
      241 tcctcggggt acatccgctc ctcggaggcc tcccagccca tggtcttctt ctgcattacg
      301 gggccgtcgg aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa cacgccgtcc
      361 tgcagggagg agtcctggtt aacgtgaata atgccgccgt cctcgaagtt catcacgcgc
      421 tcccacctga agccctcggg gaaggacagc ttgaagtagt cggggatgtc ggctgggtgc
      481 ttaatgtaga ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc
      541 agggggccac ccttggtcac cttcagctta gcggtctgaa aggcctcgta ggggcggccc
      601 tcgccctcgc cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg
      661 aagcgcatga actccttgat gatggccatg ttattctcct cgcccttgct caccattctc
      721 gagctcggat ccttatcgtc atcgtcgtac agatcccgac ccatttgctg tccaccagtc
      781 atgctagcca taccatgatg atgatgatga tgagaacccc ccatggttaa ttcctcctgt
      841 tagcccaaaa aacgggtatg gagaaacagt aaagagttgc gataaaaagc gtcaggtagg
      901 atccgctaat cttatggata aaaatgctat ggcatagcaa agtgtgacgc cgtgcaaata
      961 atcaatgtgg aacttt
//
