LOCUS       sequence-223958-         702 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Bacterial expression of E.coli DHFR N23PP and G51PEKN mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 702)
  AUTHORS   Liu CT, Hanoian P, French JB, Pringle TH, Hammes-Schiffer S, 
            Benkovic SJ
  TITLE     Functional significance of evolving protein sequence in 
            dihydrofolate reductase from bacteria to humans.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Jun 18;110(25):10159-64. doi: 
            10.1073/pnas.1307130110. Epub 2013 Jun 3.
  PUBMED    23733948
REFERENCE   2  (bases 1 to 702)
  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.1307130110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-06-18- 18"; volume: "110"; issue: "25"; pages: 
            "10159-64"
COMMENT     Sequence Label: pET22b-ecDHFR-N23PP/G51PEKN
FEATURES             Location/Qualifiers
     source          1..702
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             2..24
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             570..587
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(640..658)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      654..701
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ttttgtttaa ctttaagaag gagatataca tatgatcagt ctgattgcgg cgctagcggt
       61 agatcgcgtt atcggcatgg aaaacgccat gccatggccg ccgctgcctg ccgatctcgc
      121 ctggtttaaa cgcaacacct taaataaacc cgtgattatg gggcgccata cctgggaatc
      181 aatcccagaa aaaaacaggc ctttgcccgg ccgcaaaaat attatcctca gcagtcaacc
      241 cgggaccgat gatcgggtta cctgggttaa atcggtcgac gaagccatcg cggcgtgtgg
      301 tgacgtacca gaaatcatgg tgattggcgg cggacgcgtt tatgaacagt tcttgccaaa
      361 agcgcaaaaa ctgtatctga cgcatatcga tgcagaagtg gaaggcgaca cccattttcc
      421 ggattacgag ccggatgact gggaatcggt attcagcgaa ttccacgatg ctgatgcgca
      481 gaactcgcat agctattgtt tcgaaatcct cgagcgtcgt taaggatccg aattcgagct
      541 ccgtcgacaa gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg
      601 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
      661 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gc
//
