Coding

Part:BBa_K3930032

Designed by: Thomas Gaudin   Group: iGEM21_Toulouse_INSA-UPS   (2021-10-08)
Revision as of 12:49, 15 October 2021 by Ambre-Jousselin (Talk | contribs)


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 adenylyltransferase , which inactivates spectinomycin by adding a 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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