LOCUS       sequence-80246-e         847 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 847)
  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 847)
  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: pET21a-BetI
FEATURES             Location/Qualifiers
     source          1..847
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     59..78
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        59..77
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    78..102
                     /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)."
     RBS             117..139
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             158..175
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..208
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             827..844
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
       61 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
      121 tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc atcacagcag
      181 cggcctggaa gttctgttcc aggggcccca tatgccgaaa ctgggtatgc agagcattcg
      241 tcgtcgtcag ctgattgatg caaccctgga agcaattaat gaagttggta tgcatgatgc
      301 aaccattgca cagattgcac gtcgtgccgg tgttagcacc ggtattatta gccattattt
      361 ccgcgataaa aacggtctgc tggaagcaac catgcgtgat attaccagcc agctgcgtga
      421 tgcagttctg aatcgtctgc atgcactgcc gcagggtagc gcagaacagc gtctgcaggc
      481 aattgttggt ggtaattttg atgaaaccca ggttagcagc gcagcaatga aagcatggct
      541 ggcattttgg gcaagcagca tgcatcagcc gatgctgtat cgtctgcagc aggttagcag
      601 tcgtcgtctg ctgagcaatc tggttagcga atttcgtcgt gaactgcctc gtgaacaggc
      661 acaagaggca ggttatggtc tggcagcact gattgatggt ctgtggctgc gtgcagcact
      721 gagcggtaaa ccgctggata aaacccgtgc aaatagcctg acccgtcatt ttatcaccca
      781 gcatctgccg accgattaag gatccaagct tgcggccgca ctcgagcacc accaccacca
      841 ccactga
//
