LOCUS       sequence-80177-e         830 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 830)
  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 830)
  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-KstR
FEATURES             Location/Qualifiers
     source          1..830
                     /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"
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 cccatatgat gggtgcaacc ctgcctcgta ttgccgaagt
      241 tcgtgatgca gcagaaccga gcagtgatga acagcgtgca cgtcatgttc gtatgctgga
      301 agcagcagca gaactgggca ccgaaaaaga actgagccgt gttcagatgc atgaagttgc
      361 aaaacgtgcc ggtgttgcaa ttggcaccct gtatcgttat tttccgagca aaacccacct
      421 gtttgttgca gttatggttg agcagattga tcagattggt gatagctttg ccaaacatca
      481 ggttcagagc gcaaatccgc aggatgcagt ttatgaagtt ctggttcgtg caacccgtgg
      541 tctgctgcgt cgtccggcac tgagcaccgc aatgctgcag agcagcagca ccgcaaatgt
      601 tgcaaccgtt ccggatgttg gtaaaattga tcgtggtttt cgccagatta ttctggatgc
      661 agcaggtatt gaaaatccga ccgaagaaga taataccggt ctgcgtctgc tgatgcagct
      721 gtggtttggt gttattcaga gctgtctgaa tggtcgtatt agcattccgg atgccgaata
      781 tgatattcgt aaaggttgtg atctgctgct ggttaatctg agccgtcatt
//
