LOCUS       sequence-67748-e         840 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GFP reporter protein fused to three mutated GSK3 phosphorylation 
            sites .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 840)
  AUTHORS   Taelman VF, Dobrowolski R, Plouhinec JL, Fuentealba LC, Vorwald PP, 
            Gumper I, Sabatini DD, De Robertis EM
  TITLE     Wnt signaling requires sequestration of glycogen synthase kinase 3 
            inside multivesicular endosomes.
  JOURNAL   Cell. 2010 Dec 23. 143(7):1136-48.
  PUBMED    21183076
REFERENCE   2  (bases 1 to 840)
  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: "Cell. 2010 
            Dec 23. 143(7):1136-48."
COMMENT     Sequence Label: pCS2 GFP-GSK3mut-MAPK
FEATURES             Location/Qualifiers
     source          1..840
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      2..11
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             11..34
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(35..37,38..748)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(35..37,38..748)
                     /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(77..98)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(338..357)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     685..706
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 cgccaccatg gattacaagg atgacgacga taaggtgagc aagggcgagg agctgttcac
       61 cggggtggtg cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt
      121 gtccggcgag ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac
      181 caccggcaag ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca
      241 gtgcttcagc cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc
      301 cgaaggctac gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg
      361 cgccgaggtg aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga
      421 cttcaaggag gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa
      481 cgtctatatc atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca
      541 caacatcgag gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg
      601 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa
      661 agaccccaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
      721 cactctcggc atggacgagc tgtacaagaa atcgaattca gctgccgctg ccgctgccgc
      781 tgccgctgcc gctgccgctc ctgcttctcc tgctgccgct gccgctgccg ctgcccctcc
//
