LOCUS       sequence-77128-e         960 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 960)
  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 960)
  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: pVRa11_987
FEATURES             Location/Qualifiers
     source          1..960
                     /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             825..842
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(895..913)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      909..956
                     /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 atatgatgag cgatagtccg cagaaactgg
      241 gtcgtaatga atggaatgcc tatatggata aagtgaaagc caaagatcgt gaagcctttg
      301 catttgtgtt tcgtttttat gcaccgaaac tgaaacagtt cgcctataaa catgtgggta
      361 atgaacaggt tgccatggaa atggttcaag aaaccatggc caccgtttgg cagaaagcac
      421 atctgtatga tggtaaaaaa agcgcactga gcacctggat ttataccatt attcgcaacc
      481 tgtgctttga tctgctgcgt aaacagaaag gtaaagaact gcatatccac tccgatgata
      541 tttggccgag cgaatattat ccgcctgata tggttgatca ctatagtccg gaacaggata
      601 tgctgaaaga acaggtggtg aaatttctgg atatcctgcc gaaaaatcag cgtgatgttc
      661 tgcaggcagt ttatctggaa gaactgccgc atcagcaggt tgcagaactg tttgatattc
      721 cgctgggcac cgttaaaagc cgtctgcgtc tggcagttga aaaactgcgt catagcatgc
      781 ataccgaaca gctgtaagga tccaagcttg cggccgcact cgagcaccac caccaccacc
      841 actgagatcc ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg
      901 agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga
//
