LOCUS       sequence-471330-         993 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Cytosolic expression of mEmerald.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 993)
  AUTHORS   Goldman, C., Kareva, T., Sarrafha, L., Schuldt, B.R., Sahasrabudhe, 
            A., Ahfeldt, T.,, Blanchard, J.W
  TITLE     Genetically encoded and modular subcellular organelle probes reveal 
            dysfunction in lysosomes and mitochondria driven by PRKN knockout
  JOURNAL   iScience 28 (2025)
REFERENCE   2  (bases 1 to 993)
  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: "iScience 28
            (2025)"
COMMENT     Sequence Label: pFUW mEmerald-P2A-PuroR
FEATURES             Location/Qualifiers
     source          1..993
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      1..10
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(7..9,10..12,13..723)
                     /codon_start=1
                     /product="brightly fluorescent monomeric variant of GFP"
                     /label=mEmerald
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(52..73)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(313..332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     660..681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             748..804
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(805..824)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
ORIGIN
        1 gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat cctggtcgag
       61 ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc
      121 acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg
      181 cccaccctcg tgaccacctt gacctacggc gtgcagtgct tcgcccgcta ccccgaccac
      241 atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca ggagcgcacc
      301 atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt cgagggcgac
      361 accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg caacatcctg
      421 gggcacaagc tggagtacaa ctacaacagc cacaaggtct atatcaccgc cgacaagcag
      481 aagaacggca tcaaggtgaa cttcaagacc cgccacaaca tcgaggacgg cagcgtgcag
      541 ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct gctgcccgac
      601 aaccactacc tgagcaccca gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac
      661 atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga cgagctgtac
      721 aaggatctgc agtctagagg atctggtgcc acgaatttct cactactcaa gcaggccggt
      781 gatgtcgagg aaaaccctgg tcctatgacc gagtacaagc ccacggtgcg cctcgccacc
      841 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
      901 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
      961 ttcctcacgc gcgtcgggct cgacatcggc aag
//
