LOCUS       sequence-194614-        1230 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of unreactive version of SpyTag002 fused to 
            maltose binding protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1230)
  AUTHORS   Keeble AH, Banerjee A, Reddington SC, Ferla MP, Howarth M, Khairil 
            Anuar INA
  TITLE     Evolving accelerated amidation by SpyTag/SpyCatcher to analyze 
            membrane dynamics.
  JOURNAL   Angew Chem Int Ed Engl. 2017 Oct 10. doi: 10.1002/anie.201707623.
  PUBMED    29024296
REFERENCE   2  (bases 1 to 1230)
  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: "Angew Chem 
            Int Ed Engl. 2017 Oct 10. doi: 10.1002/anie.201707623."
COMMENT     Sequence Label: pET28a-SpyTag002 DA-MBP
FEATURES             Location/Qualifiers
     source          1..1230
                     /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             58..99
                     /codon_start=1
                     /product="improved SpyTag variant that shows rapid reaction
                     with SpyCatcher002 (Keeble et al., 2017)"
                     /label=SpyTag002
                     /translation="VPTIVMVAAYKRYK"
     CDS             118..1215
                     /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     1189..1212
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
ORIGIN
        1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagcgtg
       61 cctactatcg tgatggtggc cgcctacaag cgttacaagg gtagtggtga aagtggtaaa
      121 atcgaagaag gtaaactggt aatctggatt aacggcgata aaggctataa cggtctcgct
      181 gaagtcggta agaaattcga gaaagatacc ggaattaaag tcaccgttga gcatccggat
      241 aaactggaag agaaattccc acaggttgcg gcaactggcg atggccctga cattatcttc
      301 tgggcacacg accgctttgg tggctacgct caatctggcc tgttggctga aatcaccccg
      361 gacaaagcgt tccaggacaa gctgtatccg tttacctggg atgccgtacg ttacaacggc
      421 aagctgattg cttacccgat cgctgttgaa gcgttatcgc tgatttataa caaagatctg
      481 ctgccgaacc cgccaaaaac ctgggaagag atcccggcgc tggataaaga actgaaagcg
      541 aaaggtaaga gcgcgctgat gttcaacctg caagaaccgt acttcacctg gccgctgatt
      601 gctgctgacg ggggttatgc gttcaagtat gaaaacggca agtacgacat taaagacgtg
      661 ggcgtggata acgctggcgc gaaagcgggt ctgaccttcc tggttgacct gattaaaaac
      721 aaacacatga atgcagacac cgattactcc atcgcagaag ctgcctttaa taaaggcgaa
      781 acagcgatga ccatcaacgg cccgtgggca tggtccaaca tcgacaccag caaagtgaat
      841 tatggtgtaa cggtactgcc gaccttcaag ggtcaaccat ccaaaccgtt cgttggcgtg
      901 ctgagcgcag gtattaacgc cgccagtccg aacaaagagc tggcaaaaga gttcctcgaa
      961 aactatctgc tgactgatga aggtctggaa gcggttaata aagacaaacc gctgggtgcc
     1021 gtagcgctga agtcttacga ggaagagttg gcgaaagatc cacgtattgc cgccactatg
     1081 gaaaacgccc agaaaggtga aatcatgccg aacatcccgc agatgtccgc tttctggtat
     1141 gccgtgcgta ctgcggtgat caacgccgcc agcggtcgtc agactgtcga tgaagccctg
     1201 aaagacgcgc agactaattc gagctcgtaa
//
