LOCUS       sequence-80169-e         895 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 895)
  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 895)
  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-McbR
FEATURES             Location/Qualifiers
     source          1..895
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    72..96
                     /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             111..133
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             179..202
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             875..892
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
       61 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      121 tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct
      181 ggaagttctg ttccaggggc cccatatggc agcaagcgca agcggtaaaa gcaaaaccag
      241 tgccggtgca aatcgtcgtc gtaatcgtcc gagtccgcgt cagcgtctgc tggatagcgc
      301 aaccaacctg tttaccaccg aaggtattcg tgtgattggt attgatcgta ttctgcgtga
      361 agcagatgtt gcaaaagcaa gcctgtatag cctgtttggt agcaaagatg cactggttat
      421 tgcctatctg gaaaatctgg atcagctgtg gcgtgaagca tggcgtgaac gtaccgttgg
      481 tatgaaagat ccggaagata aaatcattgc gttctttgat cagtgcatcg aagaagaacc
      541 ggaaaaagat tttcgcggta gccattttca gaatgcagca agcgaatatc cgcgtccgga
      601 aaccgatagc gaaaaaggta ttgttgcagc agttctggaa catcgtgaat ggtgtcataa
      661 aaccctgacc gatctgctga ccgaaaaaaa tggttatccg ggtacaaccc aggcaaatca
      721 gctgctggtt tttctggatg gtggtctggc aggtagccgt ctggttcata acattagtcc
      781 gctggaaacc gcacgtgatc tggcacgtca gctgctgagc gcaccgcctg cagattatag
      841 catttaagga tccaagcttg cggccgcact cgagcaccac caccaccacc actga
//
