LOCUS       sequence-75549-e         975 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 975)
  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 975)
  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: pVRa03_1244
FEATURES             Location/Qualifiers
     source          1..975
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     55..74
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        55..73
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    74..98
                     /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             113..135
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             154..171
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             181..204
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             802..819
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(872..890)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      886..933
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 atgccggcca cgatgcgtcc ggcgtagagg atcgagatct cgatcccgcg aaattaatac
       61 gactcactat aggggaattg tgagcggata acaattcccc tctagaaata attttgttta
      121 actttaagaa ggagatatac catgggcagc agccatcatc atcatcatca cagcagcggc
      181 ctggaagttc tgttccaggg gccccatatg gcagataaac tgctgattag cgaagaagaa
      241 ctgctggaag aactgcgtga tccggcacgt cgtcgtatgg catttgaaaa agttgtgcgt
      301 ctgtataacc gcaaactgta ttggcagatt cgtcgcatgg ttctgagcca tgatgatacc
      361 gatgatctgc tgcagaacac ctttatgaaa gcatggtcag cactggatgg ttttcgtggt
      421 gaagcaaaac tgagcacctg gctgtatcgt attgcaatgt atgaaagcct gaattttctg
      481 aaaaaaaaga aaattgaaga tgatatgagc attagcatta ccgatgagaa tagctatctg
      541 ctggataatc tgaccggtga tgtgtatttt gatggtgatg aagccgaaat cacctttcag
      601 aaagcaattc tggaactgcc ggataaacag cgtctggttt ttaatctgcg ctattatgat
      661 gagatgccgt atgaaaaaat ggcagaaatt accggcacca gcgaaggtgc actgaaagca
      721 agctatcacc atgcagttaa aaaactggaa cgttttctga gtccggatga ttaaggatcc
      781 aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc tgctaacaaa
      841 gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc ataacccctt
      901 ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat atccggattg
      961 gcgaatggga cgcgc
//
