LOCUS       sequence-161324-         875 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GFP without stop codon followed by porcine teschovirus-1 2A (P2A) 
            ""self-cleaving"" peptide.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 875)
  AUTHORS   Fowler DK, Stewart S, Seredick S, Eisen JS, Stankunas K, Washbourne 
            P
  TITLE     A MultiSite Gateway Toolkit for Rapid Cloning of Vertebrate 
            Expression Constructs with Diverse Research Applications.
  JOURNAL   PLoS One. 2016 Aug 8;11(8):e0159277. doi: 
            10.1371/journal.pone.0159277. eCollection 2016.
  PUBMED    27500400
REFERENCE   2  (bases 1 to 875)
  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: "PLoS One.";
            date: "2016-08-8"; volume: "11(8):e0159277. doi"; pages: " 
            10.1371/journal.pone.0159277. eCollection 2016"
COMMENT     Sequence Label: pME-GFP-P2A
FEATURES             Location/Qualifiers
     source          1..875
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     protein_bind    71..95
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      96..105
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             109..819
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     CDS             109..819
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITLGMDELYK"
     primer_bind     complement(148..169)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(409..428)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     756..777
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 atgattttat tttgactgat agtgacctgt ttcgttgcaa caaattgatg agcaatgctt
       61 ttttataatg ccaactttgt acaaaaaagc aggctgccac catggttgag caagggcgag
      121 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      181 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      241 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      301 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      361 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      421 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
      481 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
      541 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
      601 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
      661 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
      721 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
      781 gccgccggga tcactctcgg catggacgag ctgtacaagg gatcgggatc cggagccacg
      841 aacttctctc tgttaaagca agcaggagac gtgga
//
