LOCUS       sequence-74710-e         995 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  natural linear DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 995)
  AUTHORS   Stanton BC, Nielsen AA, Tamsir A, Clancy K, Peterson T, Voigt CA
  TITLE     Genomic mining of prokaryotic repressors for orthogonal logic gates.
  JOURNAL   Nat Chem Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411.
  PUBMED    24316737
REFERENCE   2  (bases 1 to 995)
  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: "Nat Chem 
            Biol. 2013 Dec 8. doi: 10.1038/nchembio.1411."
COMMENT     Sequence Label: pRF-TetR
FEATURES             Location/Qualifiers
     source          1..995
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     RBS             38..49
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    93..111
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      complement(133..160)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(176..247)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(274..897)
                     /codon_start=1
                     /gene="tetR from transposon Tn10"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     805..824
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     protein_bind    932..951
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     promoter        complement(join(957..963,964..979,980..985))
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
ORIGIN
        1 cggtgaacag ctcctcgccc ttgctcacca tctagtattt ctcctctttc tctagtagtg
       61 ctcattatat ctatcactga tagggatgtc aatctctatc actgataggg attccaccgt
      121 acgtcgtata aacgcagaaa ggcccacccg aaggtgagcc agtgtgactc tagtagagag
      181 cgttcaccga caaacaacag ataaaacgaa aggcccagtc tttcgactga gcctttcgtt
      241 ttatttgatg cctgggggta tgggccctag ttattaggac ccactttcac atttaagttg
      301 tttttctaat ccgcatatga tcaattcaag gccgaataag aaggctggct ctgcaccttg
      361 gtgatcaaat aattcgatag cttgtcgtaa taatggcggc atactatcag tagtaggtgt
      421 ttccctttct tctttagcga cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa
      481 atgccccaca gcgctgagtg catataatgc attctctagt gaaaaacctt gttggcataa
      541 aaaggctaat tgattttcga gagtttcata ctgtttttct gtaggccgtg tacctaaatg
      601 tacttttgct ccatcgcgat gacttagtaa agcacatcta aaacttttag cgttattacg
      661 taaaaaatct tgccagcttt ccccttctaa agggcaaaag tgagtatggt gcctatctaa
      721 catctcaatg gctaaggcgt cgagcaaagc ccgcttattt tttacatgcc aatacaatgt
      781 aggctgctct acacctagct tctgggcgag tttacgggtt gttaaacctt cgattccgac
      841 ctcattaagc agctctaatg cgctgttaat cactttactt ttatctaatc tggacatcga
      901 ggcctttcct gtgtgaaaac atagtccata gaattgtgag cgctcacaat tccacacatt
      961 atacgagccg atgattaatt gtcaacacag ccaac
//
