LOCUS       sequence-77270-e         951 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 951)
  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 951)
  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: pVRa31_2963
FEATURES             Location/Qualifiers
     source          1..951
                     /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             750..767
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(820..838)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      834..881
                     /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 atatgagcga tagcaatgat attgaactgg
      241 cagccaatgg tgataaacat gcatttcgtc gtctgtatga acgttatgtt cgtggtgttt
      301 atggtatggt ttttcgtctg accgcagatc gtagcctggc agatgatctg gttcaagagg
      361 tttttgttca ggcatggcgt agcctgccga aatttgatgg tcgtgcaaaa tttagcacct
      421 ggctgtttag cctggccaat aatgttggta ttaccgaaat gcgtaaacag agccgttggt
      481 ttaaacgtaa tgaacgtgcc attgatcaca atacccagga tgaaaccgat agctcagata
      541 ttgatctggg tgaactggat aaactgatta ttcgtctgcc ggaacagacc cgtcaggtgt
      601 ttgttctgca tgcactgcag gattatagcc atcagcagat tgccgatctg ctgcatattg
      661 cagaaggcac cagcaaaagc cagttttttc gtgctaaaaa actgatgcaa gaatggctgt
      721 aaggatccaa gcttgcggcc gcactcgagc accaccacca ccaccactga gatccggctg
      781 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
      841 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
      901 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg g
//
