LOCUS       sequence-256540-         822 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpyTag003-superfolder GFP in bacterial cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 822)
  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 822)
  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-sfGFP
FEATURES             Location/Qualifiers
     source          1..822
                     /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..783
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     CDS             802..819
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 atgggacgtg gcgttcctca tattgttatg gtggacgcct acaaacgcta taaatcggga
       61 ggtggttcag gccgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
      121 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
      181 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg
      241 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
      301 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
      361 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
      421 accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg
      481 ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
      541 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
      601 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
      661 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
      721 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
      781 aaaggctccg gctccggctc ccaccatcac catcaccatt ga
//
