LOCUS       sequence-77256-e         942 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 942)
  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 942)
  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_1147
FEATURES             Location/Qualifiers
     source          1..942
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     58..77
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        58..76
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    77..101
                     /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             116..138
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             157..174
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             184..207
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             736..753
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(806..824)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      820..867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa
       61 tacgactcac tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt
      121 ttaactttaa gaaggagata taccatgggc agcagccatc atcatcatca tcacagcagc
      181 ggcctggaag ttctgttcca ggggccccat atgccgctgg ataccgatct gcatacccgt
      241 tttagcgcac tgctgcaaca gcatcgtgca attgttctga aagttgcagc aagctattgt
      301 cgtgatcgtg atgaccgtga tgaactggca caagaaattg caacccagct gtggcgtgca
      361 tttccgggtt atgatccgag ccgtcgtttt agcacctgga tgtatcgtat tgcactgaat
      421 gttgcaatta gcgatctgcg tggtcgtcgt gcagccggtg aaccgattgc actggatgca
      481 gttgttgaac gtattgcaga tccgtgtgca catgatccgg ttcgtgaaca gcaggttgca
      541 gcactgtatg catttattgc acatcagctg cctctggaac gtgccctgct gctgctgtat
      601 ctggatcagc atagctatcg tgaaattgca gatgttctgg gtattagcga aaccaatgtt
      661 gccaccaaac tgagccgtct gaaaagccgt attcgtgcag aactgtaagg atccaagctt
      721 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
      781 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      841 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
      901 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt gg
//
