LOCUS       sequence-333196-        1926 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses Maltose-binding protein-super TEV protease in bacterial 
            cytoplasm. This protease version performs well in the absence of 
            added reducing agent..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1926)
  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 1926)
  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-MBP-super TEV protease
FEATURES             Location/Qualifiers
     source          1..1926
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             1..1101
                     /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="MGIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
                     EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
                     IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
                     ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
                     TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
                     ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
                     WYAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      958..967
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1075..1098
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     CDS             1150..1161
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             1177..1197
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=7xHis
                     /translation="HHHHHHH"
     CDS             1198..1905
                     /codon_start=1
                     /product="tobacco etch virus protease (Kapust et al., 
                     2001)"
                     /label=TEV protease
                     /translation="GESLFKGPRDYNPISSTIVHLTNESDGHTTSLYGIGFGPFIITNK
                     HLFRRNNGTLVVQSLHGVFKVKNTTTLQQHLIDGRDMIIIRMPKDFPPFPQKLKFREPQ
                     REERIVLVTTNFQTKSMSSMVSDTSSTFPSGDGIFWKHWIQTKDGQCGSPLVSTRDGFI
                     VGIHSASNFTNTNNYFTSVPKNFMELLTNQEAQQWVSGWRLNADSVLWGGHKVFMDKPE
                     EPFQPVKEATQLMN"
ORIGIN
        1 atgggaatcg aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt
       61 ctcgctgaag tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat
      121 ccggataaac tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt
      181 atcttctggg cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc
      241 accccggaca aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac
      301 aacggcaagc tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa
      361 gatctgctgc cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg
      421 aaagcgaaag gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg
      481 ctgattgctg ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa
      541 gacgtgggcg tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt
      601 aaaaacaaac acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa
      661 ggcgaaacag cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa
      721 gtgaattatg gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt
      781 ggcgtgctga gcgcaggtat taacgccgcc agtccgaaca aagagctggc aaaagagttc
      841 ctcgaaaact atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg
      901 ggtgccgtag cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc
      961 accatggaaa acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc
     1021 tggtatgccg tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa
     1081 gccctgaaag acgcgcagac taattcgagc tcgaacaaca acaacaataa caataacaac
     1141 aacctcggga tcgagggaag gggagatttt gcaggtcatc atcatcatca tcatcatgga
     1201 gaaagcttgt ttaaggggcc gcgtgattac aacccgatat cgagcaccat tgttcatttg
     1261 acgaatgaat ctgatgggca cacaacatcg ttgtatggta ttggatttgg tcccttcatc
     1321 attacaaaca agcacttgtt tagaagaaat aatggaacac tggtggtcca atcactacat
     1381 ggtgtattca aggtcaagaa caccacgact ttgcaacaac acctcattga tgggagggac
     1441 atgataatta ttcgcatgcc taaggatttc ccaccatttc ctcaaaagct gaaatttaga
     1501 gagccacaaa gggaagagcg catagtcctt gtgacaacca acttccaaac taagagcatg
     1561 tctagcatgg tgtcagacac tagttcgaca ttcccttcag gagatggcat attctggaag
     1621 cattggattc aaaccaagga tgggcagtgt ggcagtccat tagtatcaac tagagatggg
     1681 ttcattgttg gtatacactc agcatcgaat ttcaccaaca caaacaatta tttcacaagc
     1741 gtgccgaaaa acttcatgga attgttgaca aatcaggagg cgcagcagtg ggttagtggt
     1801 tggcgattaa atgctgactc agtattgtgg ggaggccata aagttttcat ggacaaacct
     1861 gaagagcctt ttcagccagt taaggaagcg actcaactca tgaatcgtcg tcgccgtcgc
     1921 taataa
//
