LOCUS       sequence-256443-         807 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpyTag003-mKate2 (a far-red fluorescent protein) in 
            bacterial cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 807)
  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 807)
  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-mKate2
FEATURES             Location/Qualifiers
     source          1..807
                     /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             73..765
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="VSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIKA
                     VEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTA
                     TQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMALK
                     LVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARYC
                     DLPSKLGHR"
     CDS             784..801
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atgggacgtg gcgttcctca tattgttatg gtggacgcct acaaacgcta taaatcggga
       61 ggtggttcag gcgtgagcga gctgattaag gagaacatgc acatgaagct gtacatggag
      121 ggcaccgtga acaaccacca cttcaagtgc acatccgagg gcgaaggcaa gccctacgag
      181 ggcacccaga ccatgagaat caaggccgtc gagggcggcc ctctcccctt cgccttcgac
      241 atcctggcta ccagcttcat gtacggcagc aaaaccttca tcaaccacac ccagggcatc
      301 cccgacttct ttaagcagtc cttccctgag ggcttcacat gggagagagt caccacatac
      361 gaagacgggg gcgtgctgac cgctacccag gacaccagcc tccaggacgg ctgcctcatc
      421 tacaacgtca agatcagagg ggtgaacttc ccatccaacg gccctgtgat gcagaagaaa
      481 acactcggct gggaggcctc caccgagacc ctgtaccccg ctgacggcgg cctggaaggc
      541 agagccgaca tggccctgaa gctcgtgggc gggggccacc tgatctgcaa cttgaagacc
      601 acatacagat ccaagaaacc cgctaagaac ctcaagatgc ccggcgtcta ctatgtggac
      661 agaagactgg aaagaatcaa ggaggccgac aaagagacct acgtcgagca gcacgaggtg
      721 gctgtggcca gatactgcga cctccctagc aaactggggc acagaggctc cggctccggc
      781 tcccaccatc accatcacca ttgataa
//
