LOCUS       sequence-109635-         818 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  N4-R5-RNRV RNAP mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 818)
  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 818)
  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-RNRV
FEATURES             Location/Qualifiers
     source          1..818
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     29..47
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(71..85,86..91,92..108,109..115)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    91..107
                     /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    123..139
                     /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             158..169
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             188..205
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 gttcatgacc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
       61 cctcgagaaa tcataaaaaa tttatttgct ttgtgagcgg ataacaatta taatagattc
      121 aattgtgagc ggataacaat ttcacacaga attcattaaa gaggagaaat taactatgag
      181 aggatcgcat caccatcacc atcacggatc caacacgatt aacatcgcta agaacgactt
      241 ctctgacatc gaactggctg ctataccgtt caacactctg gctgaccatt acggtgagcg
      301 tttagctcgc gaacagttgg cccttgagca tgagtcttac gagatgggtg aagcacgctt
      361 ccgcaagatg tttgagcgtc aacttaaagc tggtgaggtt gcggataacg ctgccgccaa
      421 gcctctcatc actaccctac tccctaagat gattgcacgc atcaacgact ggtttgagga
      481 agtgaaagct aagcgcggca agcgcccgac agccttccag ttcctgcaag aaatcaagcc
      541 ggaagccgta gcgtacatca ccattaagac cactctggct tgcctaacca gtgctgacaa
      601 tacaaccgtt caggctgtag caagcgcaat cggtcgggcc attgaggacg aggctcgctt
      661 cggtcgtatc cgtgaccttg aagctaagca cttcaagaaa aacgttgagg aacaactcaa
      721 caagcgcgta gggcacgtct acaagaaagc atttatgcaa gttgtcgagg ctgacatgct
      781 ctctaagggt ctactcggtg gcgaggcgtg gtcttcgt
//
