LOCUS       sequence-208365-         902 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of E.coli DHFR D27S/Y100F mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 902)
  AUTHORS   Liu CT, Francis K, Layfield JP, Huang X, Hammes-Schiffer S, Kohen A,
            Benkovic SJ
  TITLE     Escherichia coli dihydrofolate reductase catalyzed proton and 
            hydride transfers: temporal order and the roles of Asp27 and Tyr100.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Dec 23;111(51):18231-6. doi: 
            10.1073/pnas.1415940111. Epub 2014 Dec 1.
  PUBMED    25453098
REFERENCE   2  (bases 1 to 902)
  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: 
            "10.1073/pnas.1415940111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-12-23- 23"; volume: "111"; issue: "51"; pages: "18231-6"
COMMENT     Sequence Label: pET22b-ecDHFR-D27S/Y100F
FEATURES             Location/Qualifiers
     source          1..902
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             553..570
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(623..641)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      637..684
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(808..827)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tttaacttta agaaggagat atacatatga tcagtctgat tgcggcgcta gcggtagatc
       61 gcgttatcgg catggaaaac gccatgccat ggaacctgcc tgccagtctc gcctggttta
      121 aacgcaacac cttaaataaa cccgtgatta tggggcgcca tacctgggaa tcaatcggta
      181 ggcctttgcc cggccgcaaa aatattatcc tcagcagtca acccgggacc gatgatcggg
      241 ttacctgggt taaatcggtc gacgaagcca tcgcggcgtg tggtgacgta ccagaaatca
      301 tggtgattgg cggcggacgc gtttttgaac agttcttgcc aaaagcgcaa aaactgtatc
      361 tgacgcatat cgatgcagaa gtggaaggcg acacccattt tccggattac gagccggatg
      421 actgggaatc ggtattcagc gaattccacg atgctgatgc gcagaactcg catagctatt
      481 gtttcgaaat cctcgagcgt cgttaaggat ccgaattcga gctccgtcga caagcttgcg
      541 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
      601 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
      661 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
      721 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
      781 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
      841 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
      901 gt
//
