LOCUS       sequence-137278-         853 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of SpyCatcher with a peptide tag enabling 
            site-specific biotinylation..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 853)
  AUTHORS   Veggiani G, Nakamura T, Brenner MD, Gayet RV, Yan J, Robinson CV, 
            Howarth M
  TITLE     Programmable polyproteams built using twin peptide superglues.
  JOURNAL   Proc Natl Acad Sci U S A. 2016 Jan 19. pii: 201519214.
  PUBMED    26787909
REFERENCE   2  (bases 1 to 853)
  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. 2016 Jan 19. pii: 201519214."
COMMENT     Sequence Label: AviTag-SpyCatcher
FEATURES             Location/Qualifiers
     source          1..853
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             29..46
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             53..97
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             122..142
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             149..487
                     /codon_start=1
                     /product="truncated version of the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyCatcher
                     /note="forms a covalent isopeptide bond with the SpyTag 
                     peptide (Zakeri et al., 2012)"
                     /translation="VDTLSGLSSEQGQSGDMTIEEDSATHIKFSKRDEDGKELAGATME
                     LRDSSGKTISTWISDGQVKDFYLYPGKYTFVETAAPDGYEVATAITFTVNEQGQVTVNG
                     KATKGDAHI"
     CDS             552..569
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(622..640)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      636..683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(807..826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 agaaggagat atacatatgt cgtactacca tcaccatcac catcacggca gcggcctgaa
       61 cgatattttt gaagcgcaga aaattgaatg gcatgaaggc gattacgaca tcccaacgac
      121 cgaaaacctg tattttcagg gcgccatggt tgatacctta tcaggtttat caagtgagca
      181 aggtcagtcc ggtgatatga caattgaaga agatagtgct acccatatta aattctcaaa
      241 acgtgatgag gacggcaaag agttagctgg tgcaactatg gagttgcgtg attcatctgg
      301 taaaactatt agtacatgga tttcagatgg acaagtgaaa gatttctacc tgtatccagg
      361 aaaatataca tttgtcgaaa ccgcagcacc agacggttat gaggtagcaa ctgctattac
      421 ctttacagtt aatgagcaag gtcaggttac tgtaaatggc aaagcaacta aaggtgacgc
      481 tcatatttaa atggttgatg cttgaggatc cgaattcgag ctccgtcgac aagcttgcga
      541 ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa gcccgaaagg
      601 aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta
      661 aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg gcgaatggga
      721 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
      781 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
      841 gttcgccggc ttt
//
