LOCUS       sequence-77259-e         947 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 947)
  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 947)
  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: pVRa25_1645
FEATURES             Location/Qualifiers
     source          1..947
                     /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             897..914
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
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 atatgaatcg tagcctgagc attccgattc
      241 cgagccagca gggtgttgtt ccgaaaagcg gtgttagtcc ggaaaaactg agcaattatg
      301 atctgattct gcgttgtcag gcaggtagca aaccggaacg tgcagcattt gttgaactgc
      361 tgaaacgtta tcagagccat gttgatcgtc tgctgtatca tctggcaccg gattggcagg
      421 atcgtagcga tctgagccaa gaagtttgga ttcgcgttta tcgtaatctg cagcgtctga
      481 atgatccgca gaaatttaaa ggttggctga gccgtattat caccaacctg ttttatgatg
      541 aactgcgtaa acgtaaacgt gttcgtcgtc cgctgagcct ggataatccg tttcagaccc
      601 aggatggtga agttgcatgg gatgttgcaa gtgatgatcc gagtccggat gatgatctgg
      661 caacccaaga attttatgaa catctgcgtg aagcaattgc agaactgccg gaagtttttc
      721 gtaccaccat tgtgctgcgt gaaattgaag gtctggccta tgaagaaatt gccgagatta
      781 ccggtgttag cctgggcacc gtaaaaagcc gtattgcacg tgcccgtgcc aaactgcaag
      841 agatgctgca gccgtatctg gcagattaag gatccaagct tgcggccgca ctcgagcacc
      901 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaa
//
