LOCUS       sequence-153002-        1090 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  pET-Duet for bacterial expression of Cys-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 1090)
  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 1090)
  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: pMS1088
FEATURES             Location/Qualifiers
     source          1..1090
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(1..77)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     283..302
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     390..409
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        390..408
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    409..433
                     /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             448..470
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             490..507
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             517..534
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             541..834
                     /codon_start=1
                     /gene="S. cerevisiae SMT3 (truncated)"
                     /product="cleavable ubiquitin-like protein tag"
                     /label=SUMO
                     /translation="MSDSEVNQEAKPEVKPEVKPETHINLKVSDGSSEIFFKIKKTTPL
                     RRLMEAFAKRQGKEMDSLRFLYDGIRIQADQTPEDLDMEDNDIIEAHREQIGG"
ORIGIN
        1 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
       61 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
      121 gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa
      181 ggagatggag cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca
      241 agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata
      301 ggagccagca accgcacctg tggagccggt gatgccggcc acgatgcgtc cggcgtagag
      361 gatcgagatc gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
      421 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
      481 ggcagcagtc atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagcgctagc
      541 atgtcggact cagaagtcaa tcaagaagct aagccagagg tcaagccaga agtcaagcct
      601 gagactcaca tcaatttaaa ggtgtccgat ggatcttcag agatcttctt caagatcaaa
      661 aagaccactc ctttaagaag gctgatggaa gcgttcgcta aaagacaggg taaggaaatg
      721 gactccttaa gattcttgta cgacggtatt agaattcaag ctgatcagac ccctgaagat
      781 ttggacatgg aggataacga tattattgag gctcacagag aacagattgg cggctgctct
      841 ggaatggata cctataagct ggtgattgtt ctgaatggca cgacctttac ctacacgacg
      901 gaagcagttg atgcggcgac ggcggaaaaa gtgtttaagc agtatgcgaa cgataatggt
      961 gttgacggcg aatggaccta cgcagatgct accaaaacgt tcaccgtgac ggaacgtagt
     1021 atggacacgt acaagctggt tatcgttctg aacggcacca cgtttaccta caccacggaa
     1081 gcagtggatg
//
