LOCUS       sequence-137277-         936 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 936)
  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 936)
  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..936
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(29..46)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(111..449)
                     /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             complement(456..476)
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             complement(501..545)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             complement(552..569)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             589..611
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(642..661)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(643..661)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ttcgggcttt gttagcagcc ggatctcagt ggtggtggtg gtggtgctcg agtgcggtcg
       61 caagcttgtc gacggagctc gaattcggat cctcaagcat caaccattta aatatgagcg
      121 tcacctttag ttgctttgcc atttacagta acctgacctt gctcattaac tgtaaaggta
      181 atagcagttg ctacctcata accgtctggt gctgcggttt cgacaaatgt atattttcct
      241 ggatacaggt agaaatcttt cacttgtcca tctgaaatcc atgtactaat agttttacca
      301 gatgaatcac gcaactccat agttgcacca gctaactctt tgccgtcctc atcacgtttt
      361 gagaatttaa tatgggtagc actatcttct tcaattgtca tatcaccgga ctgaccttgc
      421 tcacttgata aacctgataa ggtatcaacc atggcgccct gaaaatacag gttttcggtc
      481 gttgggatgt cgtaatcgcc ttcatgccat tcaattttct gcgcttcaaa aatatcgttc
      541 aggccgctgc cgtgatggtg atggtgatgg tagtacgaca tatgtatatc tccttcttaa
      601 agttaaacaa aattatttct agagggaaac cgttgtggtc tccctatagt gagtcgtatt
      661 aatttcgcgg gatcgagatc tcgggcagcg ttgggtcctg gccacgggtg cgcatgatcg
      721 tgctcctgtc gttgaggacc cggctaggct ggcggggttg ccttactggt tagcagaatg
      781 aatcaccgat acgcgagcga acgtgaagcg actgctgctg caaaacgtct gcgacctgag
      841 caacaacatg aatggtcttc ggtttccgtg tttcgtaaag tctggaaacg cggaagtcag
      901 cgccctgcac cattatgttc cggatctgca tcgcag
//
