LOCUS       sequence-256452-        1149 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses SpyCatcher003-superfolder GFP in bacterial cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1149)
  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 1149)
  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: SpyCatcher003-sfGFP
FEATURES             Location/Qualifiers
     source          1..1149
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             52..72
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             79..417
                     /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"
     CDS             433..1143
                     /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"
ORIGIN
        1 atgtcgtact accatcacca tcaccatcac gattacgaca tcccaacgac cgaaaacctg
       61 tattttcagg gcgccatggt aaccacctta tcaggtttat caggtgagca aggtccgtcc
      121 ggtgatatga caactgaaga agatagtgct acccatatta aattctcaaa acgtgatgag
      181 gacggccgtg agttagctgg tgcaactatg gagttgcgtg attcatctgg taaaactatt
      241 agtacatgga tttcagatgg acatgtgaag gatttctacc tgtatccagg aaaatataca
      301 tttgtcgaaa ccgcagcacc agacggttat gaggtagcaa ctccaattga atttacagtt
      361 aatgaggacg gtcaggttac tgtagatggt gaagcaactg aaggtgacgc tcatactgga
      421 tccagtggta gccgtaaagg cgaagagctg ttcactggtg tcgtccctat tctggtggaa
      481 ctggatggtg atgtcaacgg tcataagttt tccgtgcgtg gcgagggtga aggtgacgca
      541 actaatggta aactgacgct gaagttcatc tgtactactg gtaaactgcc ggtaccttgg
      601 ccgactctgg taacgacgct gacttatggt gttcagtgct ttgctcgtta tccggaccat
      661 atgaagcagc atgacttctt caagtccgcc atgccggaag gctatgtgca ggaacgcacg
      721 atttccttta aggatgacgg cacgtacaaa acgcgtgcgg aagtgaaatt tgaaggcgat
      781 accctggtaa accgcattga gctgaaaggc attgacttta aagaagacgg caatatcctg
      841 ggccataagc tggaatacaa ttttaacagc cacaatgttt acatcaccgc cgataaacaa
      901 aaaaatggca ttaaagcgaa ttttaaaatt cgccacaacg tggaggatgg cagcgtgcag
      961 ctggctgatc actaccagca aaacactcca atcggtgatg gtcctgttct gctgccagac
     1021 aatcactatc tgagcacgca aagcgttctg tctaaagatc cgaacgagaa acgcgatcat
     1081 atggttctgc tggagttcgt aaccgcagcg ggcatcacgc atggtatgga tgaactgtac
     1141 aaatgataa
//
