LOCUS       sequence-75547-e         952 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 952)
  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 952)
  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: pVRa02_2817
FEATURES             Location/Qualifiers
     source          1..952
                     /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             812..829
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(882..900)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      896..943
                     /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 gagcgaacag ctgaccgatc aggttctggt
      241 tgaacgtgtt cagaaaggtg atcagaaagc atttaatctg ctggttgtgc gctatcagca
      301 taaagttgca agcctggtta gccgttatgt tccgagcggt gatgttccgg atgttgttca
      361 agaagcattc attaaagcat atcgtgccct ggatagcttt cgtggtgata gcgcctttta
      421 tacctggctg tatcgtattg cagtgaatac cgccaaaaat tatctggttg cacagggtcg
      481 tcgtccgcct agcagtgatg ttgatgcaat tgaagccgaa aactttgaaa gcggtggtgc
      541 actgaaagaa attagcaatc cggaaaatct gatgctgagc gaagaactgc gtcagattgt
      601 ttttcgtacc attgaaagcc tgccggaaga tctgcgtatg gcaattaccc tgcgtgaact
      661 ggatggtctg agctatgaag aaattgcagc cattatggat tgtccggttg gcaccgttcg
      721 tagccgtatt tttcgtgcac gtgaagccat tgataataaa gttcagccgc tgattcgtcg
      781 ttaaggatcc aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
      841 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      901 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag ga
//
