Coding

Part:BBa_K3930032

Designed by: Thomas Gaudin   Group: iGEM21_Toulouse_INSA-UPS   (2021-10-08)


Spectinomycine 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 1032
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 7

Introduction

The SpecR cassette is composed of the promoter pAadA and the resistance gene specR (aadA). The resistance gene is based on part (BBa_K1323012), but it was codon optimized for expression into S. elongatus.

This gene codes for an aminoglycoside adenyltransferase, which inactivates spectinomycin by adding an adenyl group. Therefore, this cassette allows the selection of yeast transformants upon addition of spectinomycin into the media.

Part characterization

The part (BBa_K3930032) was used to select for genomic integration of part (BBa_K3930026). Transformants were plated onto BG11 Petri plates containing 10 μg.ml-1 spectinomycin. The Toulouse INSA UPS team managed to select transformants with this selection marker (for more details, check the (BBa_K3930026) part page).


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