LOCUS       sequence-456192-        2839 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  C terminal MBP affinity tag with PreScission Protease cleavage site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2839)
  AUTHORS   Lai Z, Flanigan SF, Boudes M, Davidovich C
  TITLE     Modular cloning of multigene vectors for the baculovirus system and 
            yeast.
  JOURNAL   J Mol Biol. 2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943.
  PUBMED    39814169
REFERENCE   2  (bases 1 to 2839)
  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: "J Mol Biol.
            2025 Jan 13:168943. doi: 10.1016/j.jmb.2025.168943."
COMMENT     Sequence Label: pM.002
FEATURES             Location/Qualifiers
     source          1..2839
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             20..43
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             80..1177
                     /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="KTEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1034..1043
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     1151..1174
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     terminator      complement(1204..1238)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1309..1968)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1883..1902
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2150..2738
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2639..2658
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcaa tcctcaggtc tggaagttct gttccagggg cccggagccg caggcaccga
       61 ttacgatatc ccaacgacca aaactgaaga aggtaaactg gtaatctgga ttaacggcga
      121 taaaggctat aacggtttgg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
      181 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
      241 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
      301 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
      361 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc
      421 gctgatttat aacaaagatt tgctgccgaa cccgccaaaa acctgggaag agatcccggc
      481 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
      541 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
      601 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
      661 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
      721 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
      781 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
      841 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
      901 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
      961 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
     1021 tccacgtatt gccgccacca tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
     1081 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
     1141 tcagactgtc gatgaagccc tgaaagacgc gcagacttaa ctcgagtggc tgagaccaga
     1201 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     1261 gtcattactg gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc
     1321 ctgccactca tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc
     1381 acaaacggca tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata
     1441 atatttgccc atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc
     1501 aaaactggtg aaactcaccc agggattggc tgaaacgaaa aacatattct caataaaccc
     1561 tttagggaaa taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag
     1621 aaactgccgg aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc
     1681 atggaaaacg gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat
     1741 tgccatacga aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg
     1801 ataaaacttg tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac
     1861 ggtctggtta taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg
     1921 ccattgggat atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt
     1981 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     2041 gtgaaagttg gaacctctta cgtgcccgat caatcatgac caaaatccct taacgtgagt
     2101 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2161 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2221 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     2281 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     2341 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     2401 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     2461 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     2521 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     2581 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     2641 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     2701 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     2761 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     2821 attctgtgga taaccgtag
//
