1.1.1.302: 2,5-diamino-6-(ribosylamino)-4(3H)-pyrimidinone 5'-phosphate reductase
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Reaction
Synonyms
2,5-diamino-6-ribitylamino-4(3H)-pyrimidinone 5'-phosphate synthase, 2,5-diamino-6-ribosylamino-4(3H)-pyrimidinone 5'-phosphate reductase, arfC, MJ0671, MJ0671 pyrimidine reductase, MjaRED, ribD2
ECTree
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Cloned
Cloned on EC 1.1.1.302 - 2,5-diamino-6-(ribosylamino)-4(3H)-pyrimidinone 5'-phosphate reductase
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expressed in Escherichia coli using a synthetic gene, the synthetic open reading frame is optimized for expression in Escherichia coli. The hypothetical open reading frame MJ0671 of Methanocaldococcus jannaschii predicts a protein of 224 amino acid residues. Because MJ0671 contains numerous codons that are poorly translated in Escherichia coli, a synthetic gene is designed that is optimized for the Escherichia coli codon usage. Approximately 31% (69 of 224) of the codons are replaced, and 14 singular restriction sites are introduced. The DNA sequence is assembled from 16 synthetic oligonucleotides by a sequence of eight PCR steps. The synthetic gene is transcribed efficiently in a recombinant Escherichia coli strain, affording approximately 30% of cellular protein
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expression in Escherichia coli