LOCUS       sequence-137149-         774 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  bacterial express ecDHFR protein for washing out TMP ligand for pBMN
            DHFR(DD)-YFP (Plasmid #29325) and pBMN YFP-DHFR(DD) (Plasmid 
            #29326).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 774)
  TITLE     Wandless lab ecDHFR plasmid
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 774)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pET15b His6-ecDHFR (WT)
FEATURES             Location/Qualifiers
     source          1..774
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(13..492)
                     /codon_start=1
                     /gene="folA"
                     /product="E. coli dihydrofolate reductase"
                     /label=ecDHFR
                     /translation="MISLIAALAVDRVIGMENAMPWNLPADLAWFKRNTLNKPVIMGRH
                     TWESIGRPLPGRKNIILSSQPGTDDRVTWVKSVDEAIAACGDVPEIMVIGGGRVYEQFL
                     PKAQKLYLTHIDAEVEGDTHFPDYEPDDWESVFSEFHDADAQNSHSYCFEILERR"
     CDS             complement(499..516)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(526..543)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             562..584
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    599..623
                     /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)."
     primer_bind     complement(623..642)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(624..642)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(726..745)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gcagccggat ccttaccgcc gctccagaat ctcaaagcaa tagctgtgag agttctgcgc
       61 atcagcatcg tggaattcgc tgaataccga ttcccagtca tccggctcgt aatccgggaa
      121 atgggtgtcg ccttccactt ctgcgtcgat atgcgtcaga tacagttttt gcgcttttgg
      181 caagaactgt tcataaacgc gaccgccgcc aatcaccatg atttctggta cgtcaccaca
      241 cgccgcgatg gcttcatcca ccgacttcac ccacgttacg cgatcgtccg tacccggttg
      301 actgctgagg ataatatttt tgcgtcctgg caacggacga ccgattgatt cccaggtatg
      361 gcggcccata atcacgggtt tatttaaggt gttgcgttta aaccaggcga gatcggcagg
      421 caggttccac ggcatggcgt tttccatgcc gataacgcga tctaccgcta acgccgcaat
      481 cagactgatc atcatatggc tgccgcgcgg caccaggccg ctgctgtgat gatgatgatg
      541 atggctgctg cccatggtat atctccttct taaagttaaa caaaattatt tctagagggg
      601 aattgttatc cgctcacaat tcccctatag tgagtcgtat taatttcgcg ggatcgagat
      661 ctcgatcctc tacgccggac gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc
      721 tggcgcctat atcgccgaca tcaccgatgg ggaagatcgg gctcgccact tcgg
//
