LOCUS       sequence-123605-        1102 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses N300S mutant of mannitol 2-dehydrogenase from Pseudomonas 
            fluorescens, codon optimized for expression in E. coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1102)
  AUTHORS   Lucas JE, Siegel JB
  TITLE     Quantitative functional characterization of conserved molecular 
            interactions in the active site of mannitol 2-dehydrogenase.
  JOURNAL   Protein Sci. 2015 Jun;24(6):936-45. doi: 10.1002/pro.2669. Epub 2015
            Apr 2.
  PUBMED    25752240
REFERENCE   2  (bases 1 to 1102)
  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: "10.1002/pro.2669"; 
            journalName: "Protein Sci"; date: "2015-06"; volume: "24"; issue: 
            "6"; pages: "936-45"
COMMENT     Sequence Label: pfMDH_N300S
FEATURES             Location/Qualifiers
     source          1..1102
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     74..93
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        74..92
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    93..117
                     /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             132..154
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 cctgtggcgc cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatcgatctc
       61 gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa caattcccct
      121 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgaaactg aacaaacaga
      181 atctgaccca gctggcaccg gaagttaaac tgcctgcata taccctggca gatacccgtc
      241 agggtattgc acatattggt gttggtggtt ttcatcgtgc acatcaggca tattataccg
      301 atgcactgat gaatacaggt gaaggtctgg attggagcat ttgcggtgtt ggtctgcgta
      361 gcgaagatcg taaagcacgt gatgatctgg caggtcagga ttacctgttt accctgtatg
      421 aactgggcga taccgatgat accgaagttc gtgttattgg tagcattagc gatatgctgc
      481 tggcagaaga tagcgcacag gcactgattg ataaactggc aagtccggaa attcgtattg
      541 ttagcctgac cattaccgaa ggtggttatt gtattgatga tagcaacggt gaatttatgg
      601 cacatctgcc gcagattcag cacgatctgg cacatccgag cagcccgaaa accgtttttg
      661 gttttatttg tgcagcactg acacagcgtc gtgcagcagg tattccggca tttaccgtta
      721 tgagctgtga taacctgccg cataatggtg cagttacccg taaagccctg ctggcatttg
      781 cagccctgca taatgcagaa ctgcatgatt ggattaaagc ccatgttagc tttccgaatg
      841 caatggttga tcgtattacc ccgatgacca gcaccgcaca tcgtctgcaa ctgcatgacg
      901 aacatggtat tgatgatgca tggcctgttg tttgtgaacc gtttgttcag tgggttctgg
      961 aagataaatt tgttaatggt cgtccggcat gggaaaaagt tggtgttcag tttaccgatg
     1021 atgttacccc gtatgaagaa atgaaaattg gtctgctgag cggtagccat ctggcactga
     1081 cctatctggg ttttctgaaa gg
//
