Difference between revisions of "Part:BBa K5143005"
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<h1>Description</h1> | <h1>Description</h1> | ||
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| − | This backbone plasmid was linearized by PCR and used to clone various genetic elements with a view to integrating them into the V chromosome of Saccharomyces cerevisiae BY4741 yeast, which lacks the ura3 gene. To do this, it has homology sequences at the locus of interest (accession number GSE94851 on SGD), so that the recombination cassette can be integrated there (release of the cassette of interest by XhoI digestion of the plasmid). <br> | + | This backbone plasmid was linearized by PCR and used to clone various genetic elements with a view to integrating them into the V chromosome of <i>Saccharomyces cerevisiae</i> BY4741 yeast, which lacks the ura3 gene. To do this, it has homology sequences at the locus of interest (accession number GSE94851 on SGD), so that the recombination cassette can be integrated there (release of the cassette of interest by XhoI digestion of the plasmid). <br> |
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| − | In addition, this backbone plasmid possesses a high-copy-number colE1 origin of replication, a selection marker for Beta-lactamase production conferring ampicillin resistance (to select colonies following transformation into E. coli DHA alpha), an auxotrophy marker ura3 (to select yeasts that have integrated the fragment of interest) and the homogeneity zones described above. <br> | + | In addition, this backbone plasmid possesses a high-copy-number colE1 origin of replication, a selection marker for Beta-lactamase production conferring ampicillin resistance (to select colonies following transformation into <i>E. coli</i> DHA alpha), an auxotrophy marker ura3 (to select yeasts that have integrated the fragment of interest) and the homogeneity zones described above. <br> |
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| − | It was used to create 3 plasmids: <a href="https://parts.igem.org/Part:BBa_K5143025">plasmid D</a> | + | It was used to create 3 plasmids: <a href="https://parts.igem.org/Part:BBa_K5143025">plasmid D</a> and <a href="https://parts.igem.org/Part:BBa_K5143027">plasmid venus/ruby</a> <br><br> |
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This part has been synthesized in order to have the URA3 gene and the Ura3'/Ura5' homologies region. | This part has been synthesized in order to have the URA3 gene and the Ura3'/Ura5' homologies region. | ||
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Latest revision as of 13:01, 28 September 2024
backbone for integrating genetic elements into the Saccharomyces cerevisiae genome
Description
This backbone plasmid was linearized by PCR and used to clone various genetic elements with a view to integrating them into the V chromosome of Saccharomyces cerevisiae BY4741 yeast, which lacks the ura3 gene. To do this, it has homology sequences at the locus of interest (accession number GSE94851 on SGD), so that the recombination cassette can be integrated there (release of the cassette of interest by XhoI digestion of the plasmid).
In addition, this backbone plasmid possesses a high-copy-number colE1 origin of replication, a selection marker for Beta-lactamase production conferring ampicillin resistance (to select colonies following transformation into E. coli DHA alpha), an auxotrophy marker ura3 (to select yeasts that have integrated the fragment of interest) and the homogeneity zones described above.
It was used to create 3 plasmids: plasmid D and plasmid venus/ruby
Construction
This part has been synthesized in order to have the URA3 gene and the Ura3'/Ura5' homologies region.