Difference between revisions of "Part:BBa K3930020"
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<partinfo>BBa_K3930020 SequenceAndFeatures</partinfo> | <partinfo>BBa_K3930020 SequenceAndFeatures</partinfo> | ||
− | This cassette | + | This cassette enables selection of yeast transformants upon addition of G418 into the media. The G418 sequence is codon optimized for expression into <i>S. cerevisiae</i>. |
<html> | <html> | ||
<h2>Introduction</h2> | <h2>Introduction</h2> | ||
− | <p>The NeoR cassette is composed | + | <p>The NeoR cassette is composed of the promoter CYC1, the resistance gene <i>neoR</i> and the transcription terminator of CYC1. The resistance gene is based on part (BBa_K1313004), but it was codon optimized for expression into <i>S. cerevisiae</i>. Indeed, this part was characterized for bacterial selection, but the <i>neoR</i> gene has a broad range of action against plenty of different aminoglycosides </p> |
− | <p>This gene codes for a | + | <p>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. </p> |
<h3>Part characterization</h3> | <h3>Part characterization</h3> | ||
− | <p> The part (BBa_K3930020) was used to select | + | <p> 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 eucaryote organism). The Toulouse INSA UPS team managed to select transformants with this selection marker (for more details, check the (BBa_K3930002) part page) </p> |
</html> | </html> |
Revision as of 15:13, 15 October 2021
G418 selective cassette
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 873
- 1000INCOMPATIBLE 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 eucaryote organism). The Toulouse INSA UPS team managed to select transformants with this selection marker (for more details, check the (BBa_K3930002) part page)