LOCUS       sequence-75542-e         950 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 950)
  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 950)
  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: pVRa01_3473
FEATURES             Location/Qualifiers
     source          1..950
                     /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"
     CDS             785..802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(855..873)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      869..916
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
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 gtttgaaaaa agtgatgaac agctgatcga
      241 aaaagccctg aaaggtcata aaaaaagctg gtttagcctg ctgaaacgct atgaaaaacc
      301 ggtgtatcat tatggtctgc gtatgaccgg taataatgca gatgcactgg atctgctgca
      361 ggatgttttt attagcgtgt ttcgtaatct ggcaacctat cgtggtgatg gtagctttaa
      421 aggttggctg tttcgtattg cccattatcg ttgcatcgaa ttttatcgtc gtaaacgtcc
      481 gatgcagggt tttgatgatg caccggatat tccgtgtgat accgcaccgg ttgatcagta
      541 tattgaaaac acccagcgta atgcacagct ggttagcgcc atgcaacaac tgccgctggc
      601 acagaaaagc gttattgaac tgaaattttt cggccacttc acctttgaag aaattgcaga
      661 acagaccggt atgagcgtta ataccgcaaa aagccgtctg tatagcgcac tgagcaaaat
      721 gaaaaccctg atggaagtgg attacgttga actgtaagga tccaagcttg cggccgcact
      781 cgagcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa aggaagctga
      841 gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt
      901 cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
//
