LOCUS       sequence-125089-         973 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  construct for expression of OmpT protease in the pET28a plasmid 
            using the restriction sites NcoI and EcoRI..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 973)
  AUTHORS   Wu C, Tran JC, Zamdborg L, Durbin KR, Li M, Ahlf DR, Early BP, 
            Thomas PM, Sweedler JV, Kelleher NL
  TITLE     A protease for 'middle-down' proteomics.
  JOURNAL   Nat Methods. 2012 Jun 17;9(8):822-4. doi: 10.1038/nmeth.2074.
  PUBMED    22706673
REFERENCE   2  (bases 1 to 973)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-06-17- 17"; volume: "9"; 
            issue: "8"; pages: "822-4"
COMMENT     Sequence Label: pET28-OmpT
FEATURES             Location/Qualifiers
     source          1..973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(2..79)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     285..304
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     388..407
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        388..406
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    407..431
                     /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             446..468
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cattcaccac cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt
       61 tgcgccattc gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg
      121 aagcagccca gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc
      181 aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa
      241 caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata
      301 taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag
      361 aggatcgaga tctcgatccc gcgaaattaa tacgactcac tataggggaa ttgtgagcgg
      421 ataacaattc ccctctagaa ataattttgt ttaactttaa gaaggagata taccatggct
      481 tctaccgaga ctttatcgtt tactcctgac aacataaatg cggacattag tcttggaact
      541 ctgagcggaa aaacaaaaga gcgtgtttat ctagccgaag aaggaggccg aaaagtcagt
      601 caactcgact ggaaattcaa taacgctgca attattaaag gtgcaattaa ttgggatttg
      661 atgccccaga tatctatcgg ggctgctggc tggacaactc tcggcagccg aggtggcaat
      721 atggtcgatc aggactggat ggattccagt aaccccggaa cctggacgga tgaaagtaga
      781 caccctgata cacaactcaa ttatgccaac gaatttgatc tgaatatcaa aggctggctc
      841 ctcaacgaac ccaattaccg cctgggactc atggccggat atcaggaaag ccgttatagc
      901 tttacagcca gaggtggttc ctatatctac agttctgagg agggattcag agatgatatc
      961 ggctccttcc cga
//
