LOCUS       sequence-152945-         855 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Express mutant AlkB protein (D135S) in E. coli BL21(DE3).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 855)
  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 855)
  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-D135S
FEATURES             Location/Qualifiers
     source          1..855
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     CDS             complement(29..46)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(677..694)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(704..721)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             740..762
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    777..801
                     /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(801..820)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(802..820)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 tttcgggctt tgttgcagcc ggatctcagt ggtggtggtg gtggtgctcg agttattctt
       61 ttttacctgc ctgacggaat gtcaggttgt agcggcagtc gatggtgagt ggatgaaacc
      121 ccgctttcaa cggttgaata ccgtgataaa acagccgcga ttcaccgccc cataccacca
      181 catcgccatg ttccaacaac aaacgtttga gcggatcatt acgtttcagg ccgccaaatt
      241 gaaaaatcgc gggtaagccc agagaaacag aaacaattgg cgcgcgcaga tccggttcag
      301 atttatcctg atgcagcgac agtttcgcgc caggagcgta gcggttgata agacaagcat
      361 ctggctggaa atctggatag cccgccgccg tagccgcacg ttgacataaa ttatgaaaac
      421 tctgtggcat ggcgggccac ggtttatttg tttgcggatc aatgggcgaa tagagataac
      481 cttgccgatg ggtcgtccag cccagatgcc cacagttggt catcgccacc gacatggtat
      541 atcccccggg ggtgaccatc tggcgaaacg gcgactggct ggcaacgtca ttaatatcgc
      601 ggattagttg ctccgcagcg ttaaaagcaa aacgccgtaa aattaccgct ccagccgcca
      661 gtggctcttg catatggctg ccgcgcggca ccaggccgct gctgtgatga tgatgatgat
      721 ggctgctgcc catggtatat ctccttctta aagttaaaca aaattatttc tagaggggaa
      781 ttgttatccg ctcacaattc ccctatagtg agtcgtatta atttcgcggg atcgagatct
      841 cgatcctcta cgccg
//
