LOCUS       sequence-77258-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_1643
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 atatgagcca gagcattacc gttagctggt
      241 caaccgttga tgcacgttgt ccggaagcaa gcgttcaggt tgataaactg agcaatcatg
      301 atctgattct gcgttgtcag gttggtctgc gtccggatcg tgttgcattt gcagaactgc
      361 tgcgtcgtta tcagacccag gttgatcgtg ttctgtatca tctggcaccg gattggagcg
      421 atcgtgcaga tctggcacaa gaagtttgga ttcgcgttta tcgcaatatt aatcgtctgc
      481 aagagccgag caaatttcgt ggttggctga gccgtattgc aaccaacctg ttttatgatg
      541 aactgcgtaa acgtaaacgt gttgttagtc cgctgagcct ggatgcaccg cgtagcgttg
      601 atgatggtga aatggattgg gaaattgccg gtgatacacc gggtccggaa gaggaactga
      661 ccacccgtga attttatgaa cagctgcgtg aagcaattgc cgatctgccg gaagtttttc
      721 gtaccaccat tgttctgcgt gaaattgaag gtatggccta tgaagaaatc gcagagatta
      781 ccggtgttag cctgggcacc gttaaaagtc gtattgcacg tgcccgtagt cgcctgcaga
      841 cccagctgca gacctatctg gatgcataag gatccaagct tgcggccgca ctcgagcacc
      901 accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaa
//
