LOCUS       sequence-152936-         967 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express wild type AlkB protein in E. coli BL21(DE3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 967)
  AUTHORS   Zheng G, Qin Y, Clark WC, Dai Q, Yi C, He C, Lambowitz AM, Pan T
  TITLE     Efficient and quantitative high-throughput tRNA sequencing.
  JOURNAL   Nat Methods. 2015 Sep;12(9):835-7. doi: 10.1038/nmeth.3478. Epub 
            2015 Jul 27.
  PUBMED    26214130
REFERENCE   2  (bases 1 to 967)
  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"; doi: 
            "10.1038/nmeth.3478"; journalName: "Nat Methods"; date: "2015-09"; 
            volume: "12"; issue: "9"; pages: "835-7"
COMMENT     Sequence Label: pET30a-AlkB
FEATURES             Location/Qualifiers
     source          1..967
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(36..53)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(684..701)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(711..728)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             747..769
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    784..808
                     /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)."
     primer_bind     complement(808..827)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(809..827)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(911..930)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 gatacttctt tnggctttgt tgcagccgga tctcagtggt ggtggtggtg gtgctcgagt
       61 tattcttttt tacctgcctg acggaatgtc aggttgtagc ggcagtcgat ggtgagtgga
      121 tgaaaccccg ctttcaacgg ttgaataccg tgataaaaca gccgcgattc accgccccat
      181 accaccacat cgccatgttc caacaacaaa cgtttgagcg gatcattacg tttcaggccg
      241 ccaaattgaa aaatcgcggg taagcccaga gaaacagaaa caattggcgc gcgcagatcc
      301 ggttcgtctt tatcctgatg cagcgacagt ttcgcgccag gagcgtagcg gttgataaga
      361 caagcatctg gctggaaatc tggatagccc gccgccgtag ccgcacgttg acataaatta
      421 tgaaaactct gtggcatggc gggccacggt ttatttgttt gcggatcaat gggcgaatag
      481 agataacctt gccgatgggt cgtccagccc agatgcccac agttggtcat cgccaccgac
      541 atggtatatc ccccgggggt gaccatctgg cgaaacggcg actggctggc aacgtcatta
      601 atatcgcgga ttagttgctc cgcagcgtta aaagcaaaac gccgtaaaat taccgctcca
      661 gccgccagtg gctcttgcat atggctgccg cgcggcacca ggccgctgct gtgatgatga
      721 tgatgatggc tgctgcccat ggtatatctc cttcttaaag ttaaacaaaa ttatttctag
      781 aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt tcgcgggatc
      841 gagatctcga tcctctacgc cggacgcatc gtggccggca tcaccggcgc cacaggtgcg
      901 gttgctggcg cctatatcgc cgacatcacc gatggggnaa gatcgggctc gccacttcgg
      961 gctcatg
//
