LOCUS       sequence-100028-        1833 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express pHluorin-mKate2-hLC3 (PK-hLC3) in mammalian cells for 
            monitoring autophagy.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1833)
  AUTHORS   Tanida I, Ueno T, Uchiyama Y
  TITLE     A Super-Ecliptic, pHluorin-mKate2, Tandem Fluorescent Protein-Tagged
            Human LC3 for the Monitoring of Mammalian Autophagy.
  JOURNAL   PLoS One. 2014 Oct 23;9(10):e110600. doi: 
            10.1371/journal.pone.0110600. eCollection 2014.
  PUBMED    25340751
REFERENCE   2  (bases 1 to 1833)
  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: "2014-10-23"; volume: "9(10):e110600. doi"; pages: " 
            10.1371/journal.pone.0110600. eCollection 2014"
COMMENT     Sequence Label: pEX-PK-hLC3
FEATURES             Location/Qualifiers
     source          1..1833
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..714
                     /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
                     FVTAAGITHGMDELYK"
     primer_bind     complement(31..59)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     654..674
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             733..1428
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     CDS             1456..1830
                     /codon_start=1
                     /gene="MAP1LC3B"
                     /product="human LC3 protein"
                     /label=LC3B
                     /note="Atg8 family member found on autophagosomes"
                     /translation="MPSEKTFKQRRTFEQRVEDVRLIREQHPTKIPVIIERYKGEKQLP
                     VLDKTKFLVPDHVNMSELIKIIRRRLQLNANQAFFLLVNGHSMVSVSTPISEVYESEKD
                     EDGFLYMVYASQETFGMKLSV"
ORIGIN
        1 atgagtaaag gagaagaact tttcactgga gttgtcccaa ttcttgttga attagatggt
       61 gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg aaggtgatgc aacatacgga
      121 aaacttaccc ttaaatttat ttgcactact ggaaaactac ctgttccatg gccaacactt
      181 gtcactacgt taacttatgg tgttcaatgc ttttcaagat acccagatca tatgaaacgg
      241 catgactttt tcaagagtgc catgcccgaa ggttacgtac aggaaagaac tatatttttc
      301 aaagatgacg ggaactacaa gacacgtgct gaagtcaagt ttgaaggtga tacccttgtt
      361 aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct tggacacaaa
      421 ttggaataca actataacga tcaccaggtg tacatcatgg cagacaaaca aaagaatgga
      481 atcaaagcta acttcaaaat tagacacaac attgaagatg gaggcgttca actagcagac
      541 cattatcaac aaaatactcc aattggcgat gggcccgtcc ttttaccaga caaccattac
      601 ctgtttacaa cttctactct ttcgaaagat cccaacgaaa agagagacca catggtcctt
      661 cttgagtttg taacagctgc tgggattaca catggcatgg atgaactata caaaggtgga
      721 ggaggtagat ccatggtgag cgagctgatt aaggagaaca tgcacatgaa gctgtacatg
      781 gagggcaccg tgaacaacca ccacttcaag tgcacatccg agggcgaagg caagccctac
      841 gagggcaccc agaccatgag aatcaaggcg gtcgagggcg gccctctccc cttcgccttc
      901 gacatcctgg ctaccagctt catgtacggc agcaaaacct tcatcaacca cacccagggc
      961 atccccgact tctttaagca gtccttcccc gagggcttca catgggagag agtcaccaca
     1021 tacgaagacg ggggcgtgct gaccgctacc caggacacca gcctccagga cggctgcctc
     1081 atctacaacg tcaagatcag aggggtgaac ttcccatcca acggccctgt gatgcagaag
     1141 aaaacactcg gctgggaggc ctccaccgag accctgtacc ccgctgacgg cggcctggaa
     1201 ggcagagccg acatggccct gaagctcgtg ggcgggggcc acctgatctg caacttgaag
     1261 accacataca gatccaagaa acccgctaag aacctcaaga tgcccggcgt ctactatgtg
     1321 gacagaagac tggaaagaat caaggaggcc gacaaagaga cctacgtcga gcagcacgag
     1381 gtggctgtgg ccagatactg cgacctccct agcaaactgg ggcacagagg tggaggaggt
     1441 tccggactca gatctatgcc gtcggagaag accttcaagc agcgccgcac cttcgaacaa
     1501 agagtagaag atgtccgact tattcgagag cagcatccaa ccaaaatccc ggtgataata
     1561 gaacgataca agggtgagaa gcagcttcct gttctggata aaacaaagtt ccttgtacct
     1621 gaccatgtca acatgagtga gctcatcaag ataattagaa ggcgcttaca gctcaatgct
     1681 aatcaggcct tcttcctgtt ggtgaacgga cacagcatgg tcagcgtctc cacaccaatc
     1741 tcagaggtgt atgagagtga gaaagatgaa gatggattcc tgtacatggt ctacgcctcc
     1801 caggagacgt tcgggatgaa attgtcagtg taa
//
