LOCUS       sequence-77274-e         954 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 954)
  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 954)
  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: pVRa33_375
FEATURES             Location/Qualifiers
     source          1..954
                     /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             804..821
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(874..892)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      888..935
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
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 atatgaccca ggcagcaagt gatgaaattc
      241 tgattgcacg tattgcacag ggtgatcgtc tggcaatgca ggttctgtat ggtcgtcatc
      301 atgttcgtgt ttatcgtttt ggtctgcgtc tggttcgtga tgaacaggca gcagaagatc
      361 tgattagcga agtttttctg gatgtttggc gtcaggcagg taaatttgaa ggtcgtagcg
      421 cagttagcac ctggctgctg gcaattaccc gttttaaagc actgagcgca ctgcgtcgtc
      481 gtaaagattt tgaactggat gaagatgcag ccaatgcaat tgaagatacc agcgataatc
      541 cggaaaccgt tgttcagaaa aaagatacct ctgaagcact gcgtgaatgt ctgaccggtc
      601 tgagtccgga acatcgtgaa attgttgatc tggtgtacta tcacgagaaa agcgttgaag
      661 aagtggcaga aatcgttggt attccggaaa ataccgttaa aacccgtctg ttttatgccc
      721 gtaaaaaact ggcagaactg ctgaaagcag ccggtgttga acgtggttgg ccgtaaggat
      781 ccaagcttgc ggccgcactc gagcaccacc accaccacca ctgagatccg gctgctaaca
      841 aagcccgaaa ggaagctgag ttggctgctg ccaccgctga gcaataacta gcataacccc
      901 ttggggcctc taaacgggtc ttgaggggtt ttttgctgaa aggaggaact atat
//
