LOCUS       sequence-60730-e         906 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 906)
  AUTHORS   Moroz OV, Moroz YS, Wu Y, Olsen AB, Cheng H, Mack KL, McLaughlin JM,
            Raymond EA, Zhezherya K, Roder H, Korendovych IV
  TITLE     A Single Mutation in a Regulatory Protein Produces Evolvable 
            Allosterically Regulated Catalyst of Nonnatural Reaction.
  JOURNAL   Angew Chem Int Ed Engl. 2013 Apr 29. doi: 10.1002/anie.201302339.
  PUBMED    23630096
REFERENCE   2  (bases 1 to 906)
  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: "Angew Chem 
            Int Ed Engl. 2013 Apr 29. doi: 10.1002/anie.201302339."
COMMENT     Sequence Label: pMCSG49-AlleyCat7
FEATURES             Location/Qualifiers
     source          1..906
                     /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             151..168
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             193..213
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             512..529
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(582..600)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      596..643
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(767..786)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc gatctcgatc ccgcgaaatt
       61 aatacgactc actatagggg aattgtgagc ggataacaat tcccctctag aaataatttt
      121 gtttaacttt aagaaggaga tatacatatg caccatcatc atcatcattc ttctggtgta
      181 gatctgggta ccgagaacct gtacttccaa tccatgaaag atacagatag cgaagaagaa
      241 cttagagaac agttccgtgt ggaagacaag gatggtaatg gttacattag tgctgcagaa
      301 cttcgtattg tgatgacaaa tcgtggggag aagctaacag atgaagaagt tgatgaactg
      361 attagggaaa cagacattga tggtgatggt caagtaaact atgaagagtt tgtacaacgt
      421 atgacagcga agtgaaaggg tgatccggca ttggaagtgg ataacggatc cgaattcgag
      481 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
      541 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
      601 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
      661 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      721 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      781 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      841 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      901 gatggt
//
