LOCUS       sequence-256446-         834 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpyTag003-mClover3 (a green fluorescent protein) in 
            bacterial cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 834)
  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 834)
  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: SpyTag003-mClover3
FEATURES             Location/Qualifiers
     source          1..834
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             7..54
                     /codon_start=1
                     /product="improved SpyTag variant that reacts with 
                     SpyCatcher003 at a rate approaching the diffusion limit 
                     (Keeble et al., 2019)"
                     /label=SpyTag003
                     /translation="RGVPHIVMVDAYKRYK"
     CDS             join(73..75,76..786)
                     /codon_start=1
                     /product="bright green-yellow fluorescent protein derived 
                     from mClover2, with improved photostability (Bajar et al., 
                     2003)"
                     /label=mClover3
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHYVYITADKQKNCIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(115..136)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     723..744
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             811..828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atgggacgtg gcgttcctca tattgttatg gtggacgcct acaaacgcta taaatcggga
       61 ggtggttcag gcgtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc
      121 gagctggacg gcgacgtaaa cggccacaag ttcagcgtcc gcggcgaggg cgagggcgat
      181 gccaccaacg gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc
      241 tggcccaccc tcgtgaccac cttcggctac ggcgtggcct gcttcagccg ctaccccgac
      301 cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc
      361 accatctctt tcaaggacga cggtacctac aagacccgcg ccgaggtgaa gttcgagggc
      421 gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc
      481 ctggggcaca agctggagta caacttcaac agccactacg tctatatcac ggccgacaag
      541 cagaagaact gcatcaaggc taacttcaag atccgccaca acgttgagga cggcagcgtg
      601 cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc
      661 gacaaccact acctgagcca tcagtccaag ctgagcaaag accccaacga gaagcgcgat
      721 cacatggtcc tgctggagtt cgtgaccgcc gccgggatta cacatggcat ggacgagctg
      781 tacaagggct ccggctccgg ctccggctcc caccatcacc atcaccattg ataa
//
