LOCUS       sequence-123750-        1108 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses D230A 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 1108)
  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 1108)
  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_D230A
FEATURES             Location/Qualifiers
     source          1..1108
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     70..89
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        70..88
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    89..113
                     /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             128..150
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
        1 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc
       61 ccgcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag
      121 aaataatttt gtttaacttt aagaaggaga tatacatatg aaactgaaca aacagaatct
      181 gacccagctg gcaccggaag ttaaactgcc tgcatatacc ctggcagata cccgtcaggg
      241 tattgcacat attggtgttg gtggttttca tcgtgcacat caggcatatt ataccgatgc
      301 actgatgaat acaggtgaag gtctggattg gagcatttgc ggtgttggtc tgcgtagcga
      361 agatcgtaaa gcacgtgatg atctggcagg tcaggattac ctgtttaccc tgtatgaact
      421 gggcgatacc gatgataccg aagttcgtgt tattggtagc attagcgata tgctgctggc
      481 agaagatagc gcacaggcac tgattgataa actggcaagt ccggaaattc gtattgttag
      541 cctgaccatt accgaaggtg gttattgtat tgatgatagc aacggtgaat ttatggcaca
      601 tctgccgcag attcagcacg atctggcaca tccgagcagc ccgaaaaccg tttttggttt
      661 tatttgtgca gcactgacac agcgtcgtgc agcaggtatt ccggcattta ccgttatgag
      721 ctgtgataac ctgccgcata atggtgcagt tacccgtaaa gccctgctgg catttgcagc
      781 cctgcataat gcagaactgc atgattggat taaagcccat gttagctttc cgaatgcaat
      841 ggttgcgcgt attaccccga tgaccagcac cgcacatcgt ctgcaactgc atgacgaaca
      901 tggtattgat gatgcatggc ctgttgtttg tgaaccgttt gttcagtggg ttctggaaga
      961 taaatttgtt aatggtcgtc cggcatggga aaaagttggt gttcagttta ccgatgatgt
     1021 taccccgtat gaagaaatga aaattggtct gctgaatggt agccatctgg cactgaccta
     1081 tctgggtttt ctgaaaggtt atcgtttt
//
