LOCUS       sequence-333355-        1122 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express DogCatcher-superfolder GFP protein in bacterial cytoplasm.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1122)
  AUTHORS   Keeble AH, Yadav VK, Ferla MP, Bauer CC, Chuntharpursat-Bon E, Huang
            J, Bon RS, Howarth M
  TITLE     DogCatcher allows loop-friendly protein-protein ligation.
  JOURNAL   Cell Chem Biol. 2021 Jul 28. pii: S2451-9456(21)00315-9. doi: 
            10.1016/j.chembiol.2021.07.005.
  PUBMED    34324879
REFERENCE   2  (bases 1 to 1122)
  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: "Cell Chem 
            Biol. 2021 Jul 28. pii: S2451-9456(21)00315-9. doi: 
            10.1016/j.chembiol.2021.07.005."
COMMENT     Sequence Label: pJ404-DogCatcher-sfGFP
FEATURES             Location/Qualifiers
     source          1..1122
                     /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..390
                     /codon_start=1
                     /product="engineered fragment of the RrgA adhesin of 
                     Streptococcus pneumoniae"
                     /label=DogCatcher
                     /note="forms a covalent bond with an intra-domain protein 
                     loop containing the DogTag peptide (Keeble et al., 2022)"
                     /translation="KLGEIEFIKVDKTDKKPLRGAVFSLQKQHPDYPDIYGAIDQNGTY
                     QDVRTGEDGKLTFTNLSDGKYRLIENSEPPGYKPVQNKPIVSFRIVDGEVRDVTSIVPQ
                     "
     CDS             406..1116
                     /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 gcgccatgaa actgggcgag attgaattta ttaaagtgga caaaaccgat
      121 aaaaagccgc tgcgtggtgc cgtgtttagc ctgcagaaac agcatcccga ctatcccgat
      181 atctatggcg cgattgatca gaatgggacc tatcaagatg tgcgtaccgg cgaagatggt
      241 aaactgacct ttacgaatct gagcgatggc aaatatcgcc tgattgaaaa tagcgaaccc
      301 ccgggctata aaccggtgca gaataagccg attgtgagct ttcgtattgt ggatggcgaa
      361 gtgcgtgatg tgaccagtat tgtgccgcag ggatccagtg gtagccgtaa aggcgaagag
      421 ctgttcactg gtgtcgtccc tattctggtg gaactggatg gtgatgtcaa cggtcataag
      481 ttttccgtgc gtggcgaggg tgaaggtgac gcaactaatg gtaaactgac gctgaagttc
      541 atctgtacta ctggtaaact gccggtacct tggccgactc tggtaacgac gctgacttat
      601 ggtgttcagt gctttgctcg ttatccggac catatgaagc agcatgactt cttcaagtcc
      661 gccatgccgg aaggctatgt gcaggaacgc acgatttcct ttaaggatga cggcacgtac
      721 aaaacgcgtg cggaagtgaa atttgaaggc gataccctgg taaaccgcat tgagctgaaa
      781 ggcattgact ttaaagaaga cggcaatatc ctgggccata agctggaata caattttaac
      841 agccacaatg tttacatcac cgccgataaa caaaaaaatg gcattaaagc gaattttaaa
      901 attcgccaca acgtggagga tggcagcgtg cagctggctg atcactacca gcaaaacact
      961 ccaatcggtg atggtcctgt tctgctgcca gacaatcact atctgagcac gcaaagcgtt
     1021 ctgtctaaag atccgaacga gaaacgcgat catatggttc tgctggagtt cgtaaccgca
     1081 gcgggcatca cgcatggtat ggatgaactg tacaaatgat aa
//
