LOCUS       sequence-77257-e         930 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 930)
  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 930)
  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: pVRa22_4450
FEATURES             Location/Qualifiers
     source          1..930
                     /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             741..758
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(811..829)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      825..872
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
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 atatgccgca gcagaccgat ctgcatgcac
      241 gttttagcgc actgatgcag cagcatcgtg gtattgttct gaaagttgca gcaagctatt
      301 gtcgtgatcc ggatgatcgt gcagatctgg cacaggatat tgcaacccat ctgtggcgtg
      361 catttccgag ctatgatccg catcgtcgtt ttagcacctg gatgtatcgt attgcactga
      421 atgttgcaat tagcgatctg cgtagccgtc gtcatgcacc gagcgaagtt ctggatgttg
      481 gcaccctggt tgatagcatt gcagatccgc atgctcgcga tccggcacgt gaacatcagc
      541 tgaccgcact gtataccttt attgcacagc tgcctcgtct ggaacgtgcc ctgatgctgc
      601 tgtatctgga tgatcatagt tatcgtgaaa ttgcagatgt gctgggtatt agcgaaacca
      661 atgttgccac caaactgagc cgtctgaaag cacgtattcg tgcagaactg taaggatcca
      721 agcttgcggc cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag
      781 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
      841 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg
      901 cgaatgggac gcgccctgta gcggcgcatt
//
