LOCUS       sequence-67743-e         969 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  GFP reporter protein fused to three GSK3 phosphorylation sites.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 969)
  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 969)
  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-GSK3-MAPK
FEATURES             Location/Qualifiers
     source          1..969
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     regulatory      3..12
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             12..35
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             join(36..38,39..749)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(36..38,39..749)
                     /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(78..99)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(339..358)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     686..707
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             849..872
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(937..956)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
ORIGIN
        1 ccgccaccat ggattacaag gatgacgacg ataaggtgag caagggcgag gagctgttca
       61 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      121 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      181 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
      241 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      301 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      361 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      421 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
      481 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
      541 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      601 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
      661 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      721 tcactctcgg catggacgag ctgtacaaga aatcgaattc agctgccgct tctgctgccg
      781 cttctgctgc cgcttctgct cctgcttctc ctgctgccgc tgccgctgcc gctgcccctc
      841 ctgcttacga ctacaaggac gacgatgaca agtaactcga gcctctagaa ctatagtgag
      901 tcgtattacg tagatccaga catgataaga tacattgatg agtttggaca aaccacaact
      961 agaatgcag
//
