LOCUS       sequence-148066-         968 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Toggle switch plasmid containing lacI-pdt3D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 968)
  AUTHORS   Cameron DE, Collins JJ
  TITLE     Tunable protein degradation in bacteria.
  JOURNAL   Nat Biotechnol. 2014 Dec;32(12):1276-81. doi: 10.1038/nbt.3053. Epub
            2014 Nov 17.
  PUBMED    25402616
REFERENCE   2  (bases 1 to 968)
  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: "10.1038/nbt.3053"; 
            journalName: "Nat Biotechnol"; date: "2014-12"; volume: "32"; issue:
            "12"; pages: "1276-81"
COMMENT     Sequence Label: pECJ3D
FEATURES             Location/Qualifiers
     source          1..968
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        join(105..110,111..127,128..134)
                     /label=trc promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    142..158
                     /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)."
     CDS             209..919
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(354..372)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     622..641
                     /label=mCherry-F
                     /note="mCherry, forward primer"
ORIGIN
        1 tgatcatctg cggattagaa aaacaactta aatgtgaaag tgggtcttga gagctcggac
       61 tgcttaagtc gctccatatg ctcgttcccg ggcgaaatga gctgttgaca attaatcatc
      121 cggctcgtat aatgtgtgga attgtgagcg gataacaatt tcacaccaat tgagtattaa
      181 ctatcgttca actgataggg aggcgccgat ggtgagcaag ggcgaggagg ataacatggc
      241 catcatcaag gagttcatgc gcttcaaggt tcacatggag ggctccgtga acggccacga
      301 gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct
      361 gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat
      421 gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc
      481 cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac
      541 cgtgacccag gactcctccc tgcaagacgg cgagttcatc tacaaggtga agctgcgcgg
      601 caccaacttc ccctccgacg gccccgtaat gcagaagaag actatgggct gggaggcctc
      661 ctccgagcgg atgtaccccg aggacggcgc gctgaagggc gagatcaagc agaggctgaa
      721 gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc
      781 cgtgcaactg cccggcgcgt acaacgtcaa catcaagttg gacatcacct cccacaacga
      841 ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat
      901 ggacgagctg tataagtaag atatctatcg ccctagggac cgtctcgaga gaatcaatat
      961 taatccaa
//
