LOCUS       sequence-77152-e         963 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 963)
  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 963)
  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: pVRa16_3622
FEATURES             Location/Qualifiers
     source          1..963
                     /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             828..845
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(898..916)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      912..959
                     /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 atatgcagcg taccaatagc caggatgttc
      241 tgagcacccg tgaaagccag ctgcaggcat tactgctgca gggtctggca ggcgatacct
      301 ttgcatatcg tcagtttctg accgcactgg cagcacatat tcgtggtttt ctgcgtcgtc
      361 gtctgccgca gcatccggca gaagttgaag atctgctgca agaagttctg ctggcagttc
      421 ataatgcacg tcatacctat caggcacgtc agccgctgac cgcatgggtt caggcaattg
      481 cacgttataa actggcagat catctgcgta gccatgcacg tcgtgaagca cgtcatgatc
      541 tgctggatga tgatagcgaa ctgtttgcag caagtgatga acagcctgca caggcaagcc
      601 gtgatctggg taaactgctg ggtcagctgc cggatcgtca gcgtctgccg attgttcatg
      661 ttaaactgga aggtctgagc gttgaagaaa ccgcacagat taccggtctg agcagcagcg
      721 cagttaaagt tggtattcat cgtggtctga aagcactggg caaactgatt cgtggtaaag
      781 gtcatgatga agatcgttaa ggatccaagc ttgcggccgc actcgagcac caccaccacc
      841 accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg
      901 ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc
      961 tga
//
