Coding

Part:BBa_K3930025

Designed by: Thomas Gaudin   Group: iGEM21_Toulouse_INSA-UPS   (2021-10-08)
Revision as of 20:58, 16 October 2021 by ThomasG (Talk | contribs)


Nourseothricin 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 710
    Illegal AgeI site found at 428
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 168

Introduction

The NsrR cassette is composed of the promoter TEF1, the resistance gene nsrR and the transcription terminator of CYC1. The resistance gene is based on part (BBa_K3629011), but it was codon optimized for expression into S. cerevisiae.

This gene codes for a nourseothricin acetyltransferase , which inactivates the nourseothricin by adding an acetyl group. Therefore, this cassette allows the selection of yeast transformants upon addition of nourseothricin into the media.

Part characterization

The part (BBa_K3930025) was used to select for genomic integration of part (BBa_K36290003). Transformants were plated onto YPD Petri plates containing 250 μg.ml-1 nourseothricin. The Toulouse INSA UPS team managed to select transformants with this selection marker (for more details, check the (BBa_K36290003) part page)


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