LOCUS       sequence-153006-        1058 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pET-Duet for bacterial expression of Ald-NuG2(4x)-Ald and 
            co-expression of Mycobacterium tuberculosis Formylglycine Generating
            Enzyme.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1058)
  AUTHORS   Walder R, LeBlanc MA, Van Patten WJ, Edwards D, Greenberg JA, 
            Adhikari A, Okoniewski SR, Sullan RMA, Rabuka D, Sousa MC, Perkins 
            TT
  TITLE     Rapid Characterization of a Mechanically Labile alpha-helical 
            Protein Enabled by Efficient Site-Specific Bioconjugation.
  JOURNAL   J Am Chem Soc. 2017 Jul 5. doi: 10.1021/jacs.7b02958.
  PUBMED    28677396
REFERENCE   2  (bases 1 to 1058)
  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 Am Chem 
            Soc. 2017 Jul 5. doi: 10.1021/jacs.7b02958."
COMMENT     Sequence Label: pMS1251
FEATURES             Location/Qualifiers
     source          1..1058
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(1..78)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     284..303
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     391..410
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        391..409
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    410..434
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             449..471
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             491..508
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             518..535
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             542..835
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
ORIGIN
        1 attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc gaaaggtttt
       61 gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc tgcattagga
      121 agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat ggtgcatgca
      181 aggagatgga gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac
      241 aagcgctcat gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat
      301 aggagccagc aaccgcacct gtggagccgg tgatgccggc cacgatgcgt ccggcgtaga
      361 ggatcgagat cgatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag
      421 cggataacaa ttcccctcta gaaataattt tgtttaactt taagaaggag atatacatat
      481 gggcagcagt catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagcgctag
      541 catgtcggac tcagaagtca atcaagaagc taagccagag gtcaagccag aagtcaagcc
      601 tgagactcac atcaatttaa aggtgtccga tggatcttca gagatcttct tcaagatcaa
      661 aaagaccact cctttaagaa ggctgatgga agcgttcgct aaaagacagg gtaaggaaat
      721 ggactcctta agattcttgt acgacggtat tagaattcaa gctgatcaga cccctgaaga
      781 tttggacatg gaggataacg atattattga ggctcacaga gaacagattg gcggcatggc
      841 accactgtgc acaccatcgc ggtctggaat ggatacctat aagctggtga ttgttctgaa
      901 tggcacgacc tttacctaca cgacggaagc agttgatgcg gcgacggcgg aaaaagtgtt
      961 taagcagtat gcgaacgata atggtgttga cggcgaatgg acctacgcag atgctaccaa
     1021 aacgttcacc gtgacggaac gtagtatgga cacgtaca
//
