LOCUS       sequence-419319-        2016 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express an autophagy (LC3) reporter in mammalian cells (untested).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2016)
  TITLE     Harper Lab plasmids
REFERENCE   2  (bases 1 to 2016)
  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"
COMMENT     Sequence Label: pHAGE pGK LC3Flux (HaloTag7-mGFP-LC3-STOP)
FEATURES             Location/Qualifiers
     source          1..2016
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..888
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="MEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSY
                     VWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIH
                     DWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIID
                     QNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVE
                     EYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLI
                     GSEIARWLSTLEISG"
     CDS             join(910..912,913..1623)
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(952..973)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1213..1232)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1560..1581
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atggaaatcg gtactggctt tccattcgac ccccattatg tggaagtcct gggcgagcgc
       61 atgcactacg tcgatgttgg tccgcgcgat ggcacccctg tgctgttcct gcacggtaac
      121 ccgacctcct cctacgtgtg gcgcaacatc atcccgcatg ttgcaccgac ccatcgctgc
      181 attgctccag acctgatcgg tatgggcaaa tccgacaaac cagacctggg ttatttcttc
      241 gacgaccacg tccgcttcat ggatgccttc atcgaagccc tgggtctgga agaggtcgtc
      301 ctggtcattc acgactgggg ctccgctctg ggtttccact gggccaagcg caatccagag
      361 cgcgtcaaag gtattgcatt tatggagttc atccgcccta tcccgacctg ggacgaatgg
      421 ccagaatttg cccgcgagac cttccaggcc ttccgcacca ccgacgtcgg ccgcaagctg
      481 atcatcgatc agaacgtttt tatcgagggt acgctgccga tgggtgtcgt ccgcccgctg
      541 actgaagtcg agatggacca ttaccgcgag ccgttcctga atcctgttga ccgcgagcca
      601 ctgtggcgct tcccaaacga gctgccaatc gccggtgagc cagcgaacat cgtcgcgctg
      661 gtcgaagaat acatggactg gctgcaccag tcccctgtcc cgaagctgct gttctggggc
      721 accccaggcg ttctgatccc accggccgaa gccgctcgcc tggccaaaag cctgcctaac
      781 tgcaaggctg tggacatcgg cccaggtctg aatctgctgc aagaagacaa cccggacctg
      841 atcggcagcg agatcgcgcg ctggctgtct actctggaga tttccggtgg ttcaggctcg
      901 ggatccgggg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
      961 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
     1021 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
     1081 cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta ccccgaccac
     1141 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
     1201 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
     1261 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
     1321 gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc cgacaagcag
     1381 aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag
     1441 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
     1501 aaccactacc tgagcaccca gtccaaactg agcaaagacc ccaacgagaa gcgcgatcac
     1561 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
     1621 aagggttcag gctcgggatc tgggaattcg ggtccgtccg agaagacctt caaacagcgc
     1681 cggagcttcg aacaaagagt ggaagatgtc cggctcatcc gggagcagca ccccaccaag
     1741 atcccagtga ttatagagcg atacaagggt gagaagcagc tgcccgtcct ggacaagacc
     1801 aagttccttg tacctgatca cgtgaatatg agcgaactca tcaagataat tagaaggcgc
     1861 ctgcagctca atgctaacca agccttcttc ctcctggtga atgggcacag catggtgagt
     1921 gtgtccacac ccatctctga agtgtacgag agcgagagag atgaagacgg cttcctgtac
     1981 atggtctatg cctcccagga gacgttcggg taatga
//
