Part:BBa_K2799001

nat1

This BioBrick contains the coding region of the nat1 gene encoding nourseothricin acetyltransferase, which confers resistance to nourseothricin (NTC). The sequence was kindly provided by Jakob Rendsvig and is codon-optimized for expression in Aspergillus oryzae.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Contribution:

The DTU Biobuilders demonstrated the usage of nat1 as a selection marker for A. oryzae by transforming A. oryzae RIB40 with the plasmid pDIV079 containing nat1 under the control of the strong constitutive tef1 promoter. Platings on different concentrations of NTC, 100 µg/ml, 200 µg/ml, 400 µg/ml, 800 µg/ml, showed signs of selection. This was especially true when replica-plated onto the same concentrations. Replatings of transformants identified as positive by replica-plating exhibited significantly faster growth than WT A. oryzae RIB40 when plated onto plates containing 800 µg/ml.