LOCUS       sequence-75548-e         966 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 966)
  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 966)
  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: pVRa03_1198
FEATURES             Location/Qualifiers
     source          1..966
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     53..72
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        53..71
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    72..96
                     /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             111..133
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             152..169
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             179..202
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             770..787
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(840..858)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      854..901
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 gccggccacg atgcgtccgg cgtagaggat cgagatctcg atcccgcgaa attaatacga
       61 ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat tttgtttaac
      121 tttaagaagg agatatacca tgggcagcag ccatcatcat catcatcaca gcagcggcct
      181 ggaagttctg ttccaggggc cccatatgat caacgaagat aaaatccgta aagcctgtag
      241 cagcgatcgt gaacgtggtt ttcgtctgct ggttgatagc tttcaagaac cgatctataa
      301 ctatattcgt cgtatggttg tgagccatga agatgcagaa gatgtgctgc aagaagtttt
      361 tattcgcgtt tttcgtcacc tggatcagtt tcgtaatgaa agcagcctga gcacctggat
      421 ttatcgtatt gcaaccaatg aatgtctgcg tctgctgaat agccgtaaag aagaagttat
      481 ttcagccgaa gatgtccaag aggaactgat gagcaaactg aaagcaagcg attatgtgga
      541 ttatgaagat gaactggccg tgaaattcca agaagcaatt ctgaccctgc cggaaaaaca
      601 gcgtattgtt tttaacctgc gctattatga cgagctggaa tatgaggata ttgcacgtgt
      661 tctggatagc aaagttgaaa ccctgaaagt gaactatcac tacgccaaag acaaaatcaa
      721 agaatatatc ctgaatcgtt aaggatccaa gcttgcggcc gcactcgagc accaccacca
      781 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
      841 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      901 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cggcgcatta
      961 agcgcg
//
