LOCUS       sequence-208335-         859 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of E.coli DHFR C85A/C152S/L54C mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 859)
  AUTHORS   Liu CT, Layfield JP, Stewart RJ 3rd, French JB, Hanoian P, Asbury 
            JB, Hammes-Schiffer S, Benkovic SJ
  TITLE     Probing the electrostatics of active site microenvironments along 
            the catalytic cycle for Escherichia coli dihydrofolate reductase.
  JOURNAL   J Am Chem Soc. 2014 Jul 23;136(29):10349-60. doi: 10.1021/ja5038947.
            Epub 2014 Jul 11.
  PUBMED    24977791
REFERENCE   2  (bases 1 to 859)
  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: "10.1021/ja5038947";
            journalName: "J Am Chem Soc"; date: "2014-07-23- 23"; volume: "136";
            issue: "29"; pages: "10349-60"
COMMENT     Sequence Label: pET22b-ecDHFR-dCys-L54C
FEATURES             Location/Qualifiers
     source          1..859
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             4..26
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             560..577
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(630..648)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      644..691
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(815..834)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 aattttgttt aactttaaga aggagatata catatgatca gtctgattgc ggcgctagcg
       61 gtagatcgcg ttatcggcat ggaaaacgcc atgccatgga acctgcctgc cgatctcgcc
      121 tggtttaaac gcaacacctt aaataaaccc gtgattatgg ggcgccatac ctgggaatca
      181 atcggtaggc cttgccccgg ccgcaaaaat attatcctca gcagtcaacc cgggaccgat
      241 gatcgggtta cctgggttaa atcggtcgac gaagccatcg cggcggctgg tgacgtacca
      301 gaaatcatgg tgattggcgg cggacgcgtt tatgaacagt tcttgccaaa agcgcaaaaa
      361 ctgtatctga cgcatatcga tgcagaagtg gaaggcgaca cccattttcc ggattacgag
      421 ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca gaactcgcat
      481 agctatagtt tcgaaatcct cgagcgtcgt taaggatccg aattcgagct ccgtcgacaa
      541 gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg ctaacaaagc
      601 ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat aaccccttgg
      661 ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat ccggattggc
      721 gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc
      781 gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt
      841 ctcgccacgt tcgccggct
//
