LOCUS       sequence-311018-         832 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  A plasmid for expression in E. coli to produce His-tag fusion 
            protein AlkB with Asp at position 135 mutated to Gly.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 832)
  AUTHORS   Wang Y, Katanski CD, Watkins C, Pan JN, Dai Q, Jiang Z, Pan T
  TITLE     A high-throughput screening method for evolving a demethylase enzyme
            with improved and new functionalities.
  JOURNAL   Nucleic Acids Res. 2021 Mar 18;49(5):e30. doi: 10.1093/nar/gkaa1213.
  PUBMED    33337498
REFERENCE   2  (bases 1 to 832)
  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: "Nucleic 
            Acids Res."; date: "2021-03-18"; volume: "49(5):e30. doi"; pages: " 
            10.1093/nar/gkaa1213"
COMMENT     Sequence Label: AlkB-D135G
FEATURES             Location/Qualifiers
     source          1..832
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             2..19
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             650..667
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(720..738)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      734..781
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cctggtgccg cgcggcagcc atatgcaaga gccactggcg gctggagcgg taattttacg
       61 gcgttttgct tttaacgctg cggagcaact aatccgcgat attaatgacg ttgccagcca
      121 gtcgccgttt cgccagatgg tcacccccgg gggatatacc atgtcggtgg cgatgaccaa
      181 ctgtgggcat ctgggctgga cgacccatcg gcaaggttat ctctattcgc ccattgatcc
      241 gcaaacaaat aaaccgtggc ccgccatgcc acagagtttt cataatttat gtcaacgtgc
      301 ggctacggcg gcgggctatc cagatttcca gccagatgct tgtcttatca accgctacgc
      361 tcctggcgcg aaactgtcgc tgcatcagga taaaggggaa ccggatctgc gcgcgccaat
      421 tgtttctgtt tctctgggct tacccgcgat ttttcaattt ggcggcctga aacgtaatga
      481 tccgctcaaa cgtttgttgt tggaacatgg cgatgtggtg gtatggggcg gtgaatcgcg
      541 gctgttttat cacggtattc aaccgttgaa agcggggttt catccactca ccatcgactg
      601 ccgctacaac ctgacattcc gtcaggcagg taaaaaagaa taactcgagc accaccacca
      661 ccaccactga gatccggctg ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac
      721 cgctgagcaa taactagcat aaccccttgg ggcctctaaa cgggtcttga ggggtttttt
      781 gctgaaagga ggaactatat ccggattggc gaatgggacg cgccctgtag cg
//
