LOCUS       sequence-89356-e        1116 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Expresses an insect mitochondrial arginyl-tRNA synthetase in E.coli.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 1116)
  AUTHORS   Leisinger AK, Janzen DH, Hallwachs W, Igloi GL
  TITLE     Amino acid discrimination by the nuclear encoded mitochondrial 
            arginyl-tRNA synthetase of the larva of a bruchid beetle (Caryedes 
            brasiliensis) from northwestern Costa Rica.
  JOURNAL   Insect Biochem Mol Biol. 2013 Dec;43(12):1172-80.
  PUBMED    24446543
REFERENCE   2  (bases 1 to 1116)
  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: "Insect 
            Biochem Mol Biol."; date: "2013-12"; volume: "43(12)"; pages: 
            "1172-80"
COMMENT     Sequence Label: pET-Cb2
FEATURES             Location/Qualifiers
     source          1..1116
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     63..82
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        63..81
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    82..106
                     /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             121..143
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             151..477
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             499..516
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             526..543
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             550..594
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     CDS             610..624
                     /codon_start=1
                     /product="enterokinase recognition and cleavage site"
                     /label=enterokinase site
                     /translation="DDDDK"
ORIGIN
        1 ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atcgatctcg atcccgcgaa
       61 attaatacga ctcactatag gggaattgtg agcggataac aattcccctc tagaaataat
      121 tttgtttaac tttaagaagg agatatacat atgagcgata aaattattca cctgactgac
      181 gacagttttg acacggatgt actcaaagcg gacggggcga tcctcgtcga tttctgggca
      241 gagtggtgcg gtccgtgcaa aatgatcgcc ccgattctgg atgaaatcgc tgacgaatat
      301 cagggcaaac tgaccgttgc aaaactgaac atcgatcaaa accctggcac tgcgccgaaa
      361 tatggcatcc gtggtatccc gactctgctg ctgttcaaaa acggtgaagt ggcggcaacc
      421 aaagtgggtg cactgtctaa aggtcagttg aaagagttcc tcgacgctaa cctggccggt
      481 tctggttctg gccatatgca ccatcatcat catcattctt ctggtctggt gccacgcggt
      541 tctggtatga aagaaaccgc tgctgctaaa ttcgaacgcc agcacatgga cagcccagat
      601 ctgggtaccg acgacgacga caaggccatg gcaactaaac taaaattgta cctaggaagg
      661 aaggtgatcg actgtctaca caaatacaca aaattttcgt tagcagagct ccaaccgctg
      721 attcaaatag gaaatgcaga tgagagtaaa atttccatga gtttaccttt aaacgatttg
      781 gaaaggcagt taggtgtaac gaacataacc gaggttttaa agattcagcc tgatgatatc
      841 ataaaaaaca tttacctcgt aagagatcgc gcaaacagga ggatatcatt tgaaatagat
      901 aaaactatat ttatcaaaga tgtgattgaa aactgtgcat ttccagagtt aaatttcagc
      961 ccaaaaaata ttgttgttga cttcagttca ccaaatgttg ctaaaccatt tcacttagga
     1021 cacctcaggt ctactattat aggaaatttt ataagcaacc tcaatagttt tttaaggcac
     1081 aaagtagtaa gattaaatta tgtgggaaat tggggg
//
