LOCUS       sequence-98991-e        2865 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  sfGFP and the 24xGCN4_v1 peptide array are expressed from the same 
            transcript, separated by a P2A site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2865)
  AUTHORS   Tanenbaum ME, Gilbert LA, Qi LS, Weissman JS, Vale RD
  TITLE     A Protein-Tagging System for Signal Amplification in Gene Expression
            and Fluorescence Imaging.
  JOURNAL   Cell. 2014 Oct 8. pii: S0092-8674(14)01227-6. doi: 
            10.1016/j.cell.2014.09.039.
  PUBMED    25307933
REFERENCE   2  (bases 1 to 2865)
  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. 2014 
            Oct 8. pii: S0092-8674(14)01227-6. doi: 10.1016/j.cell.2014.09.039."
COMMENT     Sequence Label: pcDNA4TO-24xGCN4_v1-P2A-sfGFP
FEATURES             Location/Qualifiers
     source          1..2865
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             55..120
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             136..201
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             217..282
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             298..363
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             379..444
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             460..525
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             541..606
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             622..687
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             703..768
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             784..849
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             865..930
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             946..1011
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     primer_bind     1027..1049
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     primer_bind     1050..1072
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             1075..1140
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1156..1221
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1237..1302
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1318..1383
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1399..1464
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1480..1545
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1561..1626
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1642..1707
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1723..1788
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1804..1869
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1885..1950
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             1966..2031
                     /codon_start=1
                     /product="GCN4 peptide that allows binding to an scFv-GFP 
                     fusion protein (Tanenbaum et al., 2014)"
                     /label=GCN4_v1
                     /translation="LLPKNYHLENEVARLKKLVGER"
     CDS             2086..2142
                     /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"
     CDS             2149..2865
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /note="mammalian codon-optimized"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKG"
     CDS             2149..2865
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYKG"
     primer_bind     complement(2176..2204)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2799..2819
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
ORIGIN
        1 atgggcgcca tcatacacgc ggccgcaatc atcggtccgg gaggttctgg aggattgttg
       61 cccaaaaact accacttgga gaacgaggtc gcaaggttga aaaaattggt cggtgagcgt
      121 ggcggctcag gcggccttct tccaaaaaat tatcatcttg aaaatgaagt tgctagactt
      181 aaaaaacttg ttggagaaag aggcgggagc ggcggcctgc tgcccaagaa ctaccacctg
      241 gagaacgagg tcgcccgcct gaagaagctg gtcggcgagc gcggtggttc tggaggtttg
      301 ctcccaaaga actaccactt ggaaaacgaa gtcgcaagat tgaagaagtt ggtcggtgaa
      361 agaggaggaa gcggaggatt attacctaaa aattatcatt tggaaaatga agttgctaga
      421 ttgaagaagt tggttggaga gagagggggg tctggaggac tactaccgaa aaactaccac
      481 ctagagaacg aggtggcccg actaaaaaag ctagtgggcg agcgcggcgg cagtggcggc
      541 ctcctcccca agaattacca cctcgaaaat gaagtagcaa ggctcaagaa gctcgtagga
      601 gaaagaggtg gttccggggg actccttcca aaaaactatc atcttgaaaa cgaagttgca
      661 cgtcttaaaa aacttgttgg cgaacgtggc ggctctggcg ggctgctgcc caagaattac
      721 cacctggaaa atgaagtggc ccgcctgaag aagctggtgg gcgaacgcgg aggcagtgga
      781 ggcctactac cgaagaacta ccacctagag aacgaggtgg cacgcctaaa gaagctagtg
      841 ggcgagcgcg gcggctcggg tggtttatta cctaagaatt atcatttaga aaatgaagta
      901 gctcgattaa agaagttagt aggtgaacgt ggtgggtctg gtggtcttct tccgaagaac
      961 taccaccttg agaacgaggt agctcgtctt aagaaacttg taggtgagcg tggtggaagc
     1021 ggtggtgggc tggcaagcca cgtttggtgc cgggagctgc atgtgtcaga ggaacttttg
     1081 ccaaagaatt atcatcttga gaacgaagtg gctcgtctta agaaactcgt aggagaaaga
     1141 ggtggcagtg gaggtctgct tcccaagaat taccacctgg aaaatgaggt agctagactg
     1201 aaaaagctcg taggcgagcg cgggggaagt gggggattgc tgccaaaaaa ttatcatttg
     1261 gagaacgaag tagcacgact aaagaaatta gttggggaga gggggggatc gggtggctta
     1321 ctcccgaaaa attatcatct cgaaaacgaa gtggctcggc taaaaaagtt agttggagag
     1381 cgtggcggtt ctggaggcct attaccaaaa aactaccacc tcgaaaatga ggtggcacgc
     1441 ttaaaaaagc tggtcggaga acggggaggc agtggtggcc tactacccaa gaattatcat
     1501 cttgagaacg aggtagcgcg tttgaagaag ctcgtcggtg aaagaggtgg ctcaggaggg
     1561 ctgctcccaa agaactatca tcttgaaaat gaggtcgctc gattaaaaaa gttagtgggc
     1621 gagaggggag gcagtggtgg gctacttcct aagaattacc acctcgaaaa cgaagtagct
     1681 cgattaaaga aattggtcgg tgagagaggt gggtcgggtg gcttactgcc gaaaaattat
     1741 catctggaaa atgaggtagc gagactaaaa aagctcgtag gtgagagagg gggttctggc
     1801 ggcttgctac caaaaaatta tcatcttgag aacgaagttg ctaggctcaa aaagctggtt
     1861 ggagagcgcg gcggctcagg cggtctgtta cccaaaaact accacttgga aaatgaggtt
     1921 gccagattga agaaactggt cggcgagcgc ggtggaagtg gaggactact tcctaaaaat
     1981 tatcatttgg agaacgaagt cgctagactg aaaaagttgg ttggggaacg aggagggtct
     2041 ggcggcaagc tagcgatcgg atcgatcgcc gcggaaggaa gcggagctac taacttcagc
     2101 ctgctgaagc aggctggaga cgtggaggag aaccctggac ctagcggtag caaaggagaa
     2161 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     2221 aaattttctg tccgtggaga gggtgaaggt gatgctacaa acggaaaact cacccttaaa
     2281 tttatttgca ctactggaaa actacctgtt ccgtggccaa cacttgtcac tactctgacc
     2341 tatggtgttc aatgcttttc ccgttatccg gatcacatga aacggcatga ctttttcaag
     2401 agtgccatgc ccgaaggtta tgtacaggaa cgcactatat ctttcaaaga tgacgggacc
     2461 tacaagacgc gtgctgaagt caagtttgaa ggtgataccc ttgttaatcg tatcgagtta
     2521 aagggtattg attttaaaga agatggaaac attcttggac acaaactcga gtacaacttt
     2581 aactcacaca atgtatacat cacggcagac aaacaaaaga atggaatcaa agctaacttc
     2641 aaaattcgcc acaacgttga agatggttcc gttcaactag cagaccatta tcaacaaaat
     2701 actccaattg gcgatggccc tgtcctttta ccagacaacc attacctgtc gacacaatct
     2761 gtcctttcga aagatcccaa cgaaaagcgt gaccacatgg tccttcttga gtttgtaact
     2821 gctgctggga ttacacatgg catggatgag ctctacaaag gttaa
//
