LOCUS       sequence-109448-         848 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  N4-R5-YRNRV RNAP mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 848)
  AUTHORS   Meyer AJ, Ellefson JW, Ellington AD
  TITLE     Directed Evolution of a Panel of Orthogonal T7 RNA Polymerase 
            Variants for in Vivo or in Vitro Synthetic Circuitry.
  JOURNAL   ACS Synth Biol. 2014 Oct 7.
  PUBMED    25279711
REFERENCE   2  (bases 1 to 848)
  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: "ACS Synth 
            Biol. 2014 Oct 7."
COMMENT     Sequence Label: pQE-WT-N4-R5-YRNRV
FEATURES             Location/Qualifiers
     source          1..848
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     31..49
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(73..87,88..93,94..110,111..117)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    93..109
                     /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)."
     protein_bind    125..141
                     /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             160..171
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             190..207
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ttgttcatga ccatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctt
       61 cacctcgaga aatcataaaa aatttatttg ctttgtgagc ggataacaat tataatagat
      121 tcaattgtga gcggataaca atttcacaca gaattcatta aagaggagaa attaactatg
      181 agaggatcgc atcaccatca ccatcacgga tccaacacga ttaacatcgc taagaacgac
      241 ttctctgaca tcgaactggc tgctatcccg ttcaacactc tggctgacca ttacggtgag
      301 cgtttagctc gcgaacagtt ggcccttgag catgagtctt acgagatggg tgaagcacgc
      361 ttccgcaaga tgtttgagcg tcaacttaaa gctggtgagg ttgcggataa cgctgccgcc
      421 aagcctctca tcactaccct actccctaag atgattgcac gcatcaacga ctggtttgag
      481 gaagtgaaag ctaagcgcgg caagcgcccg acagccttcc agttcctgca agaaatcaag
      541 ccggaagccg tagcgtacat caccattaag accactctgg cttgcctaac cagtgctgac
      601 aatacaaccg ttcaggctgt agcaagcgca atcggtcggg ccattgagga cgaggctcgc
      661 ttcggtcgta tccgtgacct tgaagctaag cacttcaaga aaaacgttga ggaacaactc
      721 aacaagcgcg tagggcacgt ctacaagaaa gcatttatgc aagttgtcga ggctgacatg
      781 ctctctaagg gtctactcgg tggcgaggcg tggtcttcgt ggcataagga agactctatt
      841 catgtagg
//
