LOCUS       sequence-357025-        1296 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses AviTag and SpyTag003 linked to maltose binding protein in 
            the bacterial cytoplasm. AviTag enables site-specific biotinylation 
            by BirA. SpyTag003 enables reaction with SpyCatcher003..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1296)
  AUTHORS   Vester SK, Rahikainen R, Khairil Anuar INA, Hills RA, Tan TK, 
            Howarth M
  TITLE     SpySwitch enables pH- or heat-responsive capture and release for 
            plug-and-display nanoassembly.
  JOURNAL   Nat Commun. 2022 Jun 28;13(1):3714. doi: 10.1038/s41467-022-31193-8.
  PUBMED    35764623
REFERENCE   2  (bases 1 to 1296)
  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: "Nat 
            Commun."; date: "2022-06-28"; volume: "13(1):3714. doi"; pages: " 
            10.1038/s41467-022-31193-8"
COMMENT     Sequence Label: AviTag-SpyTag003-MBP
FEATURES             Location/Qualifiers
     source          1..1296
                     /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..165
                     /codon_start=1
                     /product="improved SpyTag variant that reacts with 
                     SpyCatcher003 at a rate approaching the diffusion limit 
                     (Keeble et al., 2019)"
                     /label=SpyTag003
                     /translation="RGVPHIVMVDAYKRYK"
     CDS             184..1281
                     /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     1255..1278
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
ORIGIN
        1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat
       61 atgggtctga atgatatttt cgaagcgcag aaaattgaat ggcatgaagg tagcggacgt
      121 ggcgtgcctc atatcgtgat ggtggacgcc tacaagcgtt acaagggtag tggtgaaagt
      181 ggtaaaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
      241 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
      301 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
      361 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
      421 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
      481 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
      541 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
      601 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
      661 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
      721 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
      781 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
      841 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
      901 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
      961 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
     1021 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
     1081 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
     1141 actatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     1201 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     1261 gccctgaaag acgcgcagac taattcgagc tcgtaa
//
