LOCUS       sequence-197960-        1516 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Reticulophagy cargo assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1516)
  AUTHORS   Dennis PB, Mercer CA
  TITLE     The GST-BHMT assay and related assays for autophagy.
  JOURNAL   Methods Enzymol. 2009;452:97-118. doi: 
            10.1016/S0076-6879(08)03607-0.
  PUBMED    19200878
REFERENCE   2  (bases 1 to 1516)
  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: "Methods 
            Enzymol."; date: "2009"; volume: "452:97-118. doi"; pages: " 
            10.1016/S0076-6879(08)03607-0"
COMMENT     Sequence Label: GST(LSCS)GFP-cb5
FEATURES             Location/Qualifiers
     source          1..1516
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             14..667
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRAKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     625..647
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             join(683..685,686..1396)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(683..685,686..1396)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(725..746)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(986..1005)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1333..1354
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 tcgcgagcca gccatgtccc ctatactagg ttattggaaa attaagggcc ttgtgcaacc
       61 cactcgactt cttttggaat atcttgaaga aaaatatgaa gagcatttgt atgagcgcga
      121 tgaaggtgat aaatggcgag ccaaaaagtt tgaattgggt ttggagtttc ccaatcttcc
      181 ttattatatt gatggtgatg ttaaattaac acagtctatg gccatcatac gttatatagc
      241 tgacaagcac aacatgttgg gtggttgtcc aaaagagcgt gcagagattt caatgcttga
      301 aggagcggtt ttggatatta gatacggtgt ttcgagaatt gcatatagta aagactttga
      361 aactctcaaa gttgattttc ttagcaagct acctgaaatg ctgaaaatgt tcgaagatcg
      421 tttatgtcat aaaacatatt taaatggtga tcatgtaacc catcctgact tcatgttgta
      481 tgacgctctt gatgttgttt tatacatgga cccaatgtgc ctggatgcgt tcccaaaatt
      541 agtttgtttt aaaaaacgta ttgaagctat cccacaaatt gataagtact tgaaatccag
      601 caagtatata gcatggcctt tgcagggctg gcaagccacg tttggtggtg gcgaccatcc
      661 tccaaaatcg gatggcggcc gcgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      721 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      781 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      841 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      901 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      961 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
     1021 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
     1081 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
     1141 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
     1201 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
     1261 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
     1321 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
     1381 ggacgagctg tacaagtcta gaatcactac tgtcgagtct aattccagtt ggtggaccaa
     1441 ctgggtgatc ccagccatct cagccctggt ggtagctctg atgtatcgcc tctacatggc
     1501 agaagattag aagctt
//
