Part:BBa_K3930020

G418 selective cassette 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
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 873
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 722
  Illegal SapI.rc site found at 932
  

This cassette enables selection of yeast transformants upon addition of G418 into the media. The G418 sequence is codon optimized for expression into S. cerevisiae.

Introduction

The neoR cassette is composed of the promoter CYC1, the resistance gene neoR and the transcription terminator of CYC1. The resistance gene is based on part (BBa_K1313004), but it was codon optimized for expression into S. cerevisiae. Indeed, this part was characterized for bacterial selection, but the neoR gene has a broad range of action against plenty of different aminoglycosides.

This gene codes for a neomycin phosphotransferase II, which inactivates neomycin by adding a phosphate. Therefore, this cassette allows the selection of yeast transformants upon addition of neomycin into the media.

Part characterization

The part (BBa_K3930020) was used to select for genomic integration of part (BBa_K3930002). Transformants were plated onto YPD Petri plates containing 400 μg.ml-1 G418 (a neomycin analogue for eukaryotic organism). The Toulouse INSA UPS team managed to select transformants with this selection marker (for more details, check the (BBa_K3930002) part page).