Difference between revisions of "Part:BBa K3930020"
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<partinfo>BBa_K3930020 short</partinfo> | <partinfo>BBa_K3930020 short</partinfo> | ||
+ | <span class='h3bb'>Sequence and Features</span> | ||
+ | <partinfo>BBa_K3930020 SequenceAndFeatures</partinfo> | ||
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. | 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. | ||
+ | <html> | ||
+ | <h2>Introduction</h2> | ||
+ | <p>The NeoR cassette is composed by the promoter CYC1, the resistance gene <i>neoR</i> and the transcription terminator of CYC1. The resistance gene is based on the part (BBa_K1313004), but it was codon optimized for expression into <i>S.cerevisiae</i>. Indeed, this part was characterized for a 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 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. </p> | ||
+ | <h3>Part characterization</h3> | ||
+ | |||
+ | <p> 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) </p> | ||
+ | |||
+ | </html> | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
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Revision as of 06:40, 12 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 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)