LOCUS       sequence-80168-e         735 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 735)
  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 735)
  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-Orf2
FEATURES             Location/Qualifiers
     source          1..735
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     50..69
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        50..68
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    69..93
                     /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             108..130
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             149..166
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             176..199
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt aatacgactc
       61 actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt gtttaacttt
      121 aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca gcggcctgga
      181 agttctgttc caggggcccc atatgcagca gcagcatgaa gttgcagcac gtgttcgtcg
      241 tgttattgat gcagccggtg ttagcgcacg tgaatttgca cgtcgtattg ttattgatcg
      301 tcgtagcagt ccgggtccga gcaccgcacc gggtgcaagt ccgcctccga attggcctgg
      361 tagccgtaca ccggcagcat ggacctggga tggttgtagc ggtctgcgtc cggttcgtcg
      421 tcgtcgtagc accgttcgta gcccgagcgc acctcgtctg cctccggcaa gcaccgaagg
      481 tggtcgtccg ctgcagattg ttcgtgaaac cgttcgtctg attgcagaac gtggttttca
      541 tgcagttcgt gttgcagata ttgcagcagc atgtcatacc agcaccgcag caattcatta
      601 tcattttccg ggtcgtgatg aactgctgga agcagcagtt cgttggtgta tggatgaaga
      661 tacccgtcgt cgtgcagatg caaccgcagg cgcacgtcat gccggtgatg agctgcgtct
      721 gctgattgaa ctgca
//
