LOCUS       sequence-419311-        1863 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express a Mitophagy reporter in mammalian cells (untested).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1863)
  TITLE     Harper Lab plasmids
REFERENCE   2  (bases 1 to 1863)
  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 MitoFlux (pSu9-HaloTag7-mGFP-STOP)
FEATURES             Location/Qualifiers
     source          1..1863
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             256..1140
                     /codon_start=1
                     /product="modified bacterial dehalogenase that forms 
                     covalent bonds with chloroalkane derivatives"
                     /label=HaloTag(R)
                     /translation="EIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYV
                     WRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHD
                     WGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQ
                     NVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEE
                     YMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIG
                     SEIARWLSTLEISG"
     CDS             join(1147..1149,1150..1863)
                     /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(1189..1210)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1450..1469)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1797..1818
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atggcctcca ctcgtgtcct cgcctctcgc ctggcctccc agatggctgc ttccgccaag
       61 gttgcccgcc ctgctgtccg cgttgctcag gtcagcaagc gcaccatcca gactggctcc
      121 cccctccaga ccctcaagcg cacccagatg acctccatcg tcaacgccac cacccgccag
      181 gctttccaga agcgcgccta ctcttctgat ccaccggtcg ccaccggatc cgggaattcg
      241 ggctctggtt caggcgaaat cggtactggc tttccattcg acccccatta tgtggaagtc
      301 ctgggcgagc gcatgcacta cgtcgatgtt ggtccgcgcg atggcacccc tgtgctgttc
      361 ctgcacggta acccgacctc ctcctacgtg tggcgcaaca tcatcccgca tgttgcaccg
      421 acccatcgct gcattgctcc agacctgatc ggtatgggca aatccgacaa accagacctg
      481 ggttatttct tcgacgacca cgtccgcttc atggatgcct tcatcgaagc cctgggtctg
      541 gaagaggtcg tcctggtcat tcacgactgg ggctccgctc tgggtttcca ctgggccaag
      601 cgcaatccag agcgcgtcaa aggtattgca tttatggagt tcatccgccc tatcccgacc
      661 tgggacgaat ggccagaatt tgcccgcgag accttccagg ccttccgcac caccgacgtc
      721 ggccgcaagc tgatcatcga tcagaacgtt tttatcgagg gtacgctgcc gatgggtgtc
      781 gtccgcccgc tgactgaagt cgagatggac cattaccgcg agccgttcct gaatcctgtt
      841 gaccgcgagc cactgtggcg cttcccaaac gagctgccaa tcgccggtga gccagcgaac
      901 atcgtcgcgc tggtcgaaga atacatggac tggctgcacc agtcccctgt cccgaagctg
      961 ctgttctggg gcaccccagg cgttctgatc ccaccggccg aagccgctcg cctggccaaa
     1021 agcctgccta actgcaaggc tgtggacatc ggcccaggtc tgaatctgct gcaagaagac
     1081 aacccggacc tgatcggcag cgagatcgcg cgctggctgt ctactctgga gatttccggt
     1141 ctcgaggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
     1201 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
     1261 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
     1321 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
     1381 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
     1441 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1501 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1561 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1621 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1681 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1741 cactacctga gcacccagtc caaactgagc aaagacccca acgagaagcg cgatcacatg
     1801 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1861 taa
//
