LOCUS       sequence-310901-         833 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 Thr.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 833)
  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 833)
  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-D135T
FEATURES             Location/Qualifiers
     source          1..833
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             11..28
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             659..676
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(729..747)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      743..790
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
ORIGIN
        1 cagcagcggc ctggtgccgc gcggcagcca tatgcaagag ccactggcgg ctggagcggt
       61 aattttacgg cgttttgctt ttaacgctgc ggagcaacta atccgcgata ttaatgacgt
      121 tgccagccag tcgccgtttc gccagatggt cacccccggg ggatatacca tgtcggtggc
      181 gatgaccaac tgtgggcatc tgggctggac gacccatcgg caaggttatc tctattcgcc
      241 cattgatccg caaacaaata aaccgtggcc cgccatgcca cagagttttc ataatttatg
      301 tcaacgtgcg gctacggcgg cgggctatcc agatttccag ccagatgctt gtcttatcaa
      361 ccgctacgct cctggcgcga aactgtcgct gcatcaggat aaaaccgaac cggatctgcg
      421 cgcgccaatt gtttctgttt ctctgggctt acccgcgatt tttcaatttg gcggcctgaa
      481 acgtaatgat ccgctcaaac gtttgttgtt ggaacatggc gatgtggtgg tatggggcgg
      541 tgaatcgcgg ctgttttatc acggtattca accgttgaaa gcggggtttc atccactcac
      601 catcgactgc cgctacaacc tgacattccg tcaggcaggt aaaaaagaat aactcgagca
      661 ccaccaccac caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc
      721 tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag
      781 gggttttttg ctgaaaggag gaactatatc cggattggcg aatgggacgc gcc
//
