LOCUS       sequence-80211-e         858 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 858)
  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 858)
  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-Ef0113
FEATURES             Location/Qualifiers
     source          1..858
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     56..75
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        56..74
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    75..99
                     /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             114..136
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             155..172
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             182..205
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
ORIGIN
        1 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
       61 cgactcacta taggggaatt gtgagcggat aacaattccc ctctagaaat aattttgttt
      121 aactttaaga aggagatata ccatgggcag cagccatcat catcatcatc acagcagcgg
      181 cctggaagtt ctgttccagg ggccccatat gagcatcaac aaaaaagaca tcaaaaaaga
      241 agagatcatc tttaccgcca tgaaacactt tagcaaattc ggctacgaaa aaaccaaact
      301 gctggatatt gccaatgaag caaaaaccag caccaccacc gtttatagct ttttcaaaac
      361 caaacacgat ctgtatgaag ccgtgattgt gtataacctg gatattatca acagcgaact
      421 ggaaaaaatc gccgataaaa gcagcaccat cgatgaatat atcatgaaca ttattgaacg
      481 cgtgcgcctg aaaagcgata aagaacagct gctgctgaac ttttatatgc tgattagcac
      541 caatagcgtg cagattaaaa acctggaaat catcgaaaaa tttgaacgca aacgcttcag
      601 ctattacctg aaatttatcg agaacccgaa aatcaaagat cgttataccc tgctgttcat
      661 cgataacatt atcaccatct acatctacag ctacttcaac gtgttttatc gcaaaaaaat
      721 ctttctgtat ctgagcctgt atcagcagga tcgtgaaagc attgaatttg aagaggatct
      781 gatgaaatat ctgaaactgc tggtgaaaaa acgcctgctg gaaattgaat aaggatccaa
      841 gcttgcggcc gcactcga
//
