LOCUS       sequence-256625-        2421 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses EGFP-Talin head-SpyCatcher003 in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2421)
  AUTHORS   Keeble AH, Turkki P, Stokes S, Khairil Anuar INA, Rahikainen R, 
            Hytonen VP, Howarth M
  TITLE     Approaching infinite affinity through engineering of peptide-protein
            interaction.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 Dec 10. pii: 1909653116. doi: 
            10.1073/pnas.1909653116.
  PUBMED    31822621
REFERENCE   2  (bases 1 to 2421)
  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: "Proc Natl 
            Acad Sci U S A. 2019 Dec 10. pii: 1909653116. doi: 
            10.1073/pnas.1909653116."
COMMENT     Sequence Label: EGFP-Talin head-SpyCatcher003
FEATURES             Location/Qualifiers
     source          1..2421
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1..3,4..6,7..717)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(46..67)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(307..326)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     654..675
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     regulatory      766..775
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2080..2418
                     /codon_start=1
                     /product="improved SpyCatcher variant that reacts with 
                     SpyTag003 at a rate approaching the diffusion limit (Keeble
                     et al., 2019)"
                     /label=SpyCatcher003
                     /translation="VTTLSGLSGEQGPSGDMTTEEDSATHIKFSKRDEDGRELAGATME
                     LRDSSGKTISTWISDGHVKDFYLYPGKYTFVETAAPDGYEVATPIEFTVNEDGQVTVDG
                     EATEGDAHT"
ORIGIN
        1 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
       61 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      121 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      181 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
      241 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      301 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      361 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      421 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      481 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      541 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      601 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      661 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtcc
      721 ggactcagat ctcgagctca agctagcttc gaattcgcag aagctgccac catggttgcg
      781 ctttcgctga agattagcat tgggaatgtg gtgaagacga tgcaatttga gccatctacc
      841 atggtgtacg acgcctgccg catgattcgt gagcggatcc cagaggccct ggctggccct
      901 cccaacgact ttgggctctt tctgtcagat gatgacccca aaaaaggcat ctggctagag
      961 gctgggaaag ctttggacta ctacatgctc cgaaatgggg acaccatgga gtacagaaaa
     1021 aagcagagac ccctgaagat ccggatgtta gacggaacgg tgaagactat catggtggat
     1081 gactccaaga ccgtcacaga catgctcatg accatttgtg cccgaattgg tatcaccaac
     1141 catgatgagt attcactggt tcgagagctg atggaagaaa agaaagatga ggggacaggg
     1201 accctgagaa aagacaagac cctactgcga gatgaaaaga agatggagaa gctaaagcag
     1261 aagttgcaca cagacgatga gttgaactgg ctggaccatg ggcggacact gagggaacag
     1321 ggagtggaag agcatgagac gttgctgctg cggagaaagt tcttctactc agaccagaat
     1381 gtggattccc gagaccctgt acagctgaac cttctctatg tgcaggcacg agatgacatc
     1441 ctgaatggct cccatcctgt ctcctttgac aaggcctgtg aatttgcagg cttccagtgc
     1501 cagatccagt ttggacctca caatgaacag aagcacaagg ctggcttcct tgacctgaag
     1561 gacttcctgc ccaaggagta tgtgaagcag aagggagagc gtaagatctt tcaggcacac
     1621 aagaattgtg ggcagatgag tgaaattgag gccaaggtac gctacgtgaa gctggcccgt
     1681 tccctcaaga cttacggtgt ctccttcttc ctagtcaagg aaaagatgaa ggggaagaat
     1741 aaactggtgc ccaggctgtt gggcatcact aaggagtgtg tgatgcgtgt ggatgagaag
     1801 accaaggagg tgattcagga atggagcctc accaacatca aacgctgggc tgcctctccc
     1861 aagagcttta ctctggactt tggagactat caggatggct actactcagt acagacgaca
     1921 gaaggcgagc agatcgcaca gctcattgct ggctacatag atatcatcct taagaagaaa
     1981 aaaagcaagg accattttgg gctggaggga gatgaagagt ctactatgtt ggaggactca
     2041 gtgtcaccca aaaagtcaac agtccttcag ggcgcgatgg tgaccacact gtccggactg
     2101 tctggagagc agggaccatc cggcgacatg accacagagg aggattctgc cacacacatc
     2161 aagttcagca agagggacga ggacggaaga gagctggcag gagcaaccat ggagctgagg
     2221 gatagctccg gcaagaccat cagcacatgg atctccgacg gccacgtgaa ggatttctac
     2281 ctgtatcccg gcaagtacac ctttgtggag acagcagcac cagacggata tgaggtggca
     2341 acccctatcg agtttacagt gaacgaggac ggacaggtga ccgtggatgg agaggcaaca
     2401 gagggcgatg cacacaccta a
//
