LOCUS       sequence-419315-        1686 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express a ERphagy reporter in mammalian cells (untested).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1686)
  TITLE     Harper Lab plasmids
REFERENCE   2  (bases 1 to 1686)
  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 ERFlux (Bip-HaloTag7-mGFP-KDEL-STOP)
FEATURES             Location/Qualifiers
     source          1..1686
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     sig_peptide     1..54
                     /codon_start=1
                     /product="Drosophila BiP signal sequence"
                     /label=BiP signal sequence
                     /translation="MKLCILLAVVAFVGLSLG"
     CDS             58..942
                     /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(949..951,952..1662)
                     /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(991..1012)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1252..1271)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1599..1620
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgaagttat gcatattact ggccgtcgtg gcctttgttg gcctctcgct cggatccgaa
       61 atcggtactg gctttccatt cgacccccat tatgtggaag tcctgggcga gcgcatgcac
      121 tacgtcgatg ttggtccgcg cgatggcacc cctgtgctgt tcctgcacgg taacccgacc
      181 tcctcctacg tgtggcgcaa catcatcccg catgttgcac cgacccatcg ctgcattgct
      241 ccagacctga tcggtatggg caaatccgac aaaccagacc tgggttattt cttcgacgac
      301 cacgtccgct tcatggatgc cttcatcgaa gccctgggtc tggaagaggt cgtcctggtc
      361 attcacgact ggggctccgc tctgggtttc cactgggcca agcgcaatcc agagcgcgtc
      421 aaaggtattg catttatgga gttcatccgc cctatcccga cctgggacga atggccagaa
      481 tttgcccgcg agaccttcca ggccttccgc accaccgacg tcggccgcaa gctgatcatc
      541 gatcagaacg tttttatcga gggtacgctg ccgatgggtg tcgtccgccc gctgactgaa
      601 gtcgagatgg accattaccg cgagccgttc ctgaatcctg ttgaccgcga gccactgtgg
      661 cgcttcccaa acgagctgcc aatcgccggt gagccagcga acatcgtcgc gctggtcgaa
      721 gaatacatgg actggctgca ccagtcccct gtcccgaagc tgctgttctg gggcacccca
      781 ggcgttctga tcccaccggc cgaagccgct cgcctggcca aaagcctgcc taactgcaag
      841 gctgtggaca tcggcccagg tctgaatctg ctgcaagaag acaacccgga cctgatcggc
      901 agcgagatcg cgcgctggct gtctactctg gagatttccg gtctcgaggt gagcaagggc
      961 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     1021 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     1081 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     1141 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     1201 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     1261 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     1321 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     1381 tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac
     1441 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     1501 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     1561 tccaaactga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     1621 accgccgccg ggatcactct cggcatggac gagctgtaca agaattcaaa agatgagctc
     1681 tgataa
//
