LOCUS       sequence-421700-        2361 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses tandem MBPx linked to-SpyTag003 in bacterial cells. MBPx 
            is a variant of MBP (A312V I317V Delta-172-173 Delta-175-176) with 
            enhanced affinity for maltose..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 2361)
  AUTHORS   Driscoll CL, Keeble AH, Howarth MR
  TITLE     SpyMask enables combinatorial assembly of bispecific binders.
  JOURNAL   Nat Commun. 2024 Mar 16;15(1):2403. doi: 10.1038/s41467-024-46599-9.
  PUBMED    38493197
REFERENCE   2  (bases 1 to 2361)
  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: "2024-03-16"; volume: "15(1):2403. doi"; pages: " 
            10.1038/s41467-024-46599-9"
COMMENT     Sequence Label: pET21b-MBPx-MBPx-SpyTag003
FEATURES             Location/Qualifiers
     source          1..2361
                     /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             70..1158
                     /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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYGDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGETAMT
                     INGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLENYL
                     LTDEGLEAVNKDKPLGAVALKSYEEELVKDPRVAATMENAQKGEIMPNIPQMSAFWYAV
                     RTAVINAASGRQTVDEALKDAQT"
     CDS             2311..2358
                     /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"
ORIGIN
        1 atgggcagca gccatcatca tcatcatcac agcagcggcc tggtgccgcg cggcagccat
       61 atggctagca tgaaaatcga agaaggtaaa ctggtaatct ggattaacgg cgataaaggc
      121 tataacggtc tcgctgaagt cggtaagaaa ttcgagaaag ataccggaat taaagtcacc
      181 gttgagcatc cggataaact ggaagagaaa ttcccacagg ttgcggcaac tggcgatggc
      241 cctgacatta tcttctgggc acacgaccgc tttggtggct acgctcaatc tggcctgttg
      301 gctgaaatca ccccggacaa agcgttccag gacaagctgt atccgtttac ctgggatgcc
      361 gtacgttaca acggcaagct gattgcttac ccgatcgctg ttgaagcgtt atcgctgatt
      421 tataacaaag atctgctgcc gaacccgcca aaaacctggg aagagatccc ggcgctggat
      481 aaagaactga aagcgaaagg taagagcgcg ctgatgttca acctgcaaga accgtacttc
      541 acctggccgc tgattgctgc tgacgggggt tatgcgttca agtatggcga cattaaagac
      601 gtgggcgtgg ataacgctgg cgcgaaagcg ggtctgacct tcctggttga cctgattaaa
      661 aacaaacaca tgaatgcaga caccgattac tccatcgcag aagctgcctt taataaaggc
      721 gaaacagcga tgaccatcaa cggcccgtgg gcatggtcca acatcgacac cagcaaagtg
      781 aattatggtg taacggtact gccgaccttc aagggtcaac catccaaacc gttcgttggc
      841 gtgctgagcg caggtattaa cgccgccagt ccgaacaaag agctggcaaa agagttcctc
      901 gaaaactatc tgctgactga tgaaggtctg gaagcggtta ataaagacaa accgctgggt
      961 gccgtagcgc tgaagtctta cgaggaagag ttggtgaaag atccacgtgt ggccgccact
     1021 atggaaaacg cccagaaagg tgaaatcatg ccgaacatcc cgcagatgtc cgctttctgg
     1081 tatgccgtgc gtactgcggt gatcaacgcc gccagcggtc gtcagactgt cgatgaagcc
     1141 ctgaaagacg cgcagacgaa ttctagttcc ggcggatccg gaggtggatc cggaaagata
     1201 gaggagggta aactggtaat ctggattaac ggcgataaag gctataacgg tctcgctgaa
     1261 gtcggtaaga aattcgagaa agataccgga attaaagtca ccgttgagca tccggataaa
     1321 ctggaagaga aattcccaca ggttgcggca actggcgatg gccctgacat tatcttctgg
     1381 gcacacgacc gctttggtgg ctacgctcaa tctggcctgt tggctgaaat caccccggac
     1441 aaagcgttcc aggacaagct gtatccgttt acctgggatg ccgtacgtta caacggcaag
     1501 ctgattgctt acccgatcgc tgttgaagcg ttatcgctga tttataacaa agatctgctg
     1561 ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa
     1621 ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact tcacctggcc gctgattgct
     1681 gctgacgggg gttatgcgtt caagtatggc gacattaaag acgtgggcgt ggataacgct
     1741 ggcgcgaaag cgggtctgac cttcctggtt gacctgatta aaaacaaaca catgaatgca
     1801 gacaccgatt actccatcgc agaagctgcc tttaataaag gcgaaacagc gatgaccatc
     1861 aacggcccgt gggcatggtc caacatcgac accagcaaag tgaattatgg tgtaacggta
     1921 ctgccgacct tcaagggtca accatccaaa ccgttcgttg gcgtgctgag cgcaggtatt
     1981 aacgccgcca gtccgaacaa agagctggca aaagagttcc tcgaaaacta tctgctgact
     2041 gatgaaggtc tggaagcggt taataaagac aaaccgctgg gtgccgtagc gctgaagtct
     2101 tacgaggaag agttggtgaa agatccacgt gtggccgcca ctatggaaaa cgcccagaaa
     2161 ggtgaaatca tgccgaacat cccgcagatg tccgctttct ggtatgccgt gcgtactgcg
     2221 gtgatcaacg ccgccagcgg tcgtcagact gtcgatgaag ccctgaaaga cgcgcagact
     2281 aattcgagct cgggttcggg cggtagtggt cgtggcgtgc ctcatatcgt gatggtggac
     2341 gcctacaagc gttacaagta a
//
