LOCUS       sequence-77147-e         965 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 965)
  AUTHORS   Rhodius VA, Segall-Shapiro TH, Sharon BD, Ghodasara A, Orlova E, 
            Tabakh H, Burkhardt DH, Clancy K, Peterson TC, Gross CA, Voigt CA
  TITLE     Design of orthogonal genetic switches based on a crosstalk map of 
            sigmas, anti-sigmas, and promoters.
  JOURNAL   Mol Syst Biol. 2013 Oct 29;9:702. doi: 10.1038/msb.2013.58.
  PUBMED    24169405
REFERENCE   2  (bases 1 to 965)
  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: "Mol Syst 
            Biol."; date: "2013-10-29"; volume: "9:702. doi"; pages: " 
            10.1038/msb.2013.58"
COMMENT     Sequence Label: pVRa15_524
FEATURES             Location/Qualifiers
     source          1..965
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     60..79
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        60..78
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    79..103
                     /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             118..140
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             159..176
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             186..209
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             846..863
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(916..934)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 cggtgatgcc ggccacgatg cgtccggcgt agaggatcga gatctcgatc ccgcgaaatt
       61 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      121 gtttaacttt aagaaggaga tataccatgg gcagcagcca tcatcatcat catcacagca
      181 gcggcctgga agttctgttc caggggcccc atatggttgc agaacaggtt ggtcgtagtc
      241 cgcctgatcc ggttcgtgat aattcattta aacgtgaact ggttaccctg attccgcatc
      301 tgcgtgcatt tgcacgtacc ctgaccggtg atccgaccgc agcagatgat ctggcacagg
      361 atgcaatgat gaaagcatgg gatgcacgtg ccagctttca gctgggcacc aatatgaaag
      421 cctggacctt tatgattctg cgcaatcagt tctatagcga aaaacgtcgt agctggcgtc
      481 agagccaact ggatcaagaa gcagcagaac gcaccctggt tgcagttgat gatccggaag
      541 caccggttgc actggatgaa ctgcgtcagg gtctgagcat gctgcctgcc gaacagcgtg
      601 aagcactgat tctggttggt gccggtggtt ttgcctatga agaagccgct gaaatttgta
      661 attgtgcagt tggcaccgtt aaaagccgtg ttagccgtgc acgtcgtgcc ctgcaggcaa
      721 ttctggaaag cggtgcctat gatcgtgatg gtgcagcagc cggtgatgca atgcgtgcca
      781 ttctggcaga tgcagaacgt ctgagcgcac tgcgttaagg atccaagctt gcggccgcac
      841 tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg
      901 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
      961 tcttg
//
