LOCUS       sequence-77255-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: pVRa20_2913
FEATURES             Location/Qualifiers
     source          1..947
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     59..78
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        59..77
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    78..102
                     /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             117..139
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             158..175
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..208
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             788..805
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(858..876)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      872..919
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaatta
       61 atacgactca ctatagggga attgtgagcg gataacaatt cccctctaga aataattttg
      121 tttaacttta agaaggagat ataccatggg cagcagccat catcatcatc atcacagcag
      181 cggcctggaa gttctgttcc aggggcccca tatggaacgt gatcatgaag cactgctggt
      241 tgcacgtgtt gcagatggtg atcgtgcagc atttggtgaa ctgctgaaaa cccatcagca
      301 gccgctggtt cgttatgcac gtcgtatgct gagcgatccg ggtgcagcag atgatgttgt
      361 tcaagaaacc tttctgcgtc tgtggacccg tgcaggtagc tataaaagca gcagcgcacg
      421 tctgaccacc tggctgcata atattgcaca taatctgtgc gtggatagct ttcgtcgtaa
      481 tgcccgtctg agctttaccg atgatgaaag ccagctggaa gttaccaccg caggtccgga
      541 aggtgatgca gaaagcaccg atagcgcaca tcgtgttcgt accgcactgg aaggtctgcc
      601 ggaacgtcag cgtagcgcac tgctgatgtg tcattatcag ggtctgagca atcgtgaagc
      661 agcaaccgtt ctggatgtta gcgttgatgc actggaaagc ctgctggcac gtgcgcgtcg
      721 tcgtctgcgt gaagaactgc tgtcacatgt tgaaagctaa ggatccaagc ttgcggccgc
      781 actcgagcac caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc
      841 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
      901 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggattgg
//
