LOCUS       sequence-81318-e         913 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 913)
  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 913)
  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-AmeR
FEATURES             Location/Qualifiers
     source          1..913
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(36..53)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(744..767)
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             complement(777..794)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             813..835
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    850..874
                     /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(874..893)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(875..893)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcttcctttc gggctttgtt agcagccgga tctcagtggt ggtggtggtg gtgctcgagt
       61 gcggccgcaa gcttggatcc ttattttgcc agcggattac gcagacctgc atcaatcaga
      121 tcaaccagct gatcacaacg ggttgccaga ttaccaatat cttcttgtgc aatcagaacc
      181 ggatgaataa cgctggtcag tgcatgcaga acggtttctg ccagaaccgg aatatcggtt
      241 gcaatataca gacctgcttc aacaccatca cgaataattt cggccagcag ttttgcaatc
      301 acgcttttgt aataatcgcc acatttcata tcctgtttgg cggtcatcag gatcatttca
      361 aacacatacg ggtttttatt cagatcctga tgatgctgtt ccatcagatt acgtgccaga
      421 tgacgcagac gatccagcgg tgcatggctt ttatccagcg gacaaatctg acgttcaaaa
      481 acaccaatct gaccaaaacc aattgcatca accagtgcgt ttttgctgct aaaatgttta
      541 aacacatttg ccggactcat attcagtgcg cttgcaatat ctgcaatggc aactgcatta
      601 aaaccacgtt cacgaaacag ttcttcggct tttgccagaa tatcacgacg ggtttcttcg
      661 gcactacgac gcggacgacg acgaaccggc agatcgcttt tacgatcacc tttacgcacc
      721 tgatcaatgg ttttgttcat atggggcccc tggaacagaa cttccaggcc gctgctgtga
      781 tgatgatgat gatggctgct gcccatggta tatctccttc ttaaagttaa acaaaattat
      841 ttctagaggg gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc
      901 gggatcgaga tct
//
