LOCUS       sequence-333092-        1317 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses AviTag-DogTag-Maltose binding protein in bacterial 
            cytoplasm. The AviTag enables site-specific biotinylation by BirA..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1317)
  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 1317)
  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: pET28-AviTag-DogTag-MBP
FEATURES             Location/Qualifiers
     source          1..1317
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             13..30
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             40..57
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             64..108
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             118..186
                     /codon_start=1
                     /product="peptide derived from the RrgA adhesin of 
                     Streptococcus pneumoniae"
                     /label=DogTag
                     /note="forms a covalent bond with the DogCatcher protein 
                     when inserted into an intra-domain protein loop (Keeble et 
                     al., 2022)"
                     /translation="DIPATYEFTDGKHYITNEPIPPK"
     CDS             205..1302
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     primer_bind     1276..1299
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
ORIGIN
        1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat
       61 atgggtctga atgatatttt cgaagcgcag aaaattgaat ggcatgaagg tagcggagat
      121 attccggcta catacgaatt caccgatggt aaacattata tcaccaatga accgataccg
      181 ccgaaaggta gtggtgaaag tggtaaaatc gaagaaggta aactggtaat ctggattaac
      241 ggcgataaag gctataacgg tctcgctgaa gtcggtaaga aattcgagaa agataccgga
      301 attaaagtca ccgttgagca tccggataaa ctggaagaga aattcccaca ggttgcggca
      361 actggcgatg gccctgacat tatcttctgg gcacacgacc gctttggtgg ctacgctcaa
      421 tctggcctgt tggctgaaat caccccggac aaagcgttcc aggacaagct gtatccgttt
      481 acctgggatg ccgtacgtta caacggcaag ctgattgctt acccgatcgc tgttgaagcg
      541 ttatcgctga tttataacaa agatctgctg ccgaacccgc caaaaacctg ggaagagatc
      601 ccggcgctgg ataaagaact gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa
      661 gaaccgtact tcacctggcc gctgattgct gctgacgggg gttatgcgtt caagtatgaa
      721 aacggcaagt acgacattaa agacgtgggc gtggataacg ctggcgcgaa agcgggtctg
      781 accttcctgg ttgacctgat taaaaacaaa cacatgaatg cagacaccga ttactccatc
      841 gcagaagctg cctttaataa aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg
      901 tccaacatcg acaccagcaa agtgaattat ggtgtaacgg tactgccgac cttcaagggt
      961 caaccatcca aaccgttcgt tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac
     1021 aaagagctgg caaaagagtt cctcgaaaac tatctgctga ctgatgaagg tctggaagcg
     1081 gttaataaag acaaaccgct gggtgccgta gcgctgaagt cttacgagga agagttggcg
     1141 aaagatccac gtattgccgc cactatggaa aacgcccaga aaggtgaaat catgccgaac
     1201 atcccgcaga tgtccgcttt ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc
     1261 ggtcgtcaga ctgtcgatga agccctgaaa gacgcgcaga ctaattcgag ctcgtaa
//
