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 allows the selection of Yeast transformant upon addition of G418 into the media of culture. The sequence of the gene carrying the resistance is codon optimized for an expression into S.cerevisiae.

Introduction

The NeoR cassette is composed by the promoter CYC1, the resistance gene neoR and the transcription terminator of CYC1. The resistance gene is based on the part (BBa_K1313004), but it was codon optimized for expression into S.cerevisiae. Indeed, this part was characterized for a 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 inactivate the neomycin by adding a phosphate on it. Therefore, this cassette aload the selection of Yeast transformant upon addition of neomycin into the media of culture.

Part characterization

The part (BBa_K3930020) was used to select the right genomic integration of the part (BBa_K3930002). Transformants were plated onto YPD + 400 μg.ml-1 G418 (an analogue of neomycin for eucaryote organism) Petri dish. The Toulouse INSA UPS team manage to select a transformant with this selection marker (for more details, check the (BBa_K3930002) part page)