LOCUS       sequence-77254-e         946 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 946)
  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 946)
  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: pVRa20_992
FEATURES             Location/Qualifiers
     source          1..946
                     /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             822..839
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(892..910)
                     /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 atatgaatga aaccgatcct gatctggaac
      241 tgctgaaacg tattggtaat aatgatgcac aggccgttaa agaaatggtt acccgtaaac
      301 tgcctcgtct gctggcactg gcaagtcgcc tgctgggtga tgcagatgaa gcacgtgata
      361 ttgcacaaga aagttttctg cgcatttgga aacaggcagc aagctggcgt agcgaacagg
      421 cacgttttga tacctggctg catcgtgttg cactgaatct gtgttatgat cgtctgcgtc
      481 gtcgtaaaga acatgtgccg gttgatagcg aacatgcctg tgaagcactg gatacccgtc
      541 cggcaccgga tgaacagctg gaagcaagcg cacagagccg tcgtatggca caggcactgg
      601 atcagctgcc ggatcgtcag cgtgaagcaa ttgttctgca gtattatcaa gaactgagca
      661 ataccgaagc agcagcactg atgcaaatta gcgttgaagc cctggaaagc ctgctgagcc
      721 gtgcacgtcg taatctgcgt agccatctgg ccgaagcacc gggtgcagat ctgagcggtc
      781 gtcgcaaacc gtaaggatcc aagcttgcgg ccgcactcga gcaccaccac caccaccact
      841 gagatccggc tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc
      901 aataactagc ataacccctt ggggcctcta aacgggtctt gagggg
//
