Primers Design For Yeast Integration
From 2007.igem.org
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First we cloned 2XORE into Hind-Bam E.R. site of p2UG25 plasmid that has natMX4 resistence, so we create a 2XORE-natMX4 cassette,that after we can integrate in the Pho4's ORF. | First we cloned 2XORE into Hind-Bam E.R. site of p2UG25 plasmid that has natMX4 resistence, so we create a 2XORE-natMX4 cassette,that after we can integrate in the Pho4's ORF. | ||
| - | 2XORE Fv Hind | + | 2XORE Fv Hind CGCGGATCCATCAGTATTACCCTCGACCTC |
| - | 2XORE Rv Bam | + | 2XORE Rv Bam CCCAAGCTTTGTTTGCGTGTCTATAGAAG |
| - | 2XOREint Fv | + | 2XOREint Fv TCTAGATCGTCCACAAAACCGTGTATTCCCTCAGAAGTTGTACGGCCCATTGTTTGCGTGTCTATAGAAG |
| - | 2XOREint Rv | + | 2XOREint Rv CAGAAGAGAAGAGATGAGCAAAGGAGACAGAACAAGAGTAGCAGAAAGTCATCGATGAATTCGAGCTCG |
Revision as of 18:26, 18 October 2007
To create our circuite in yeast cell, we have to integrate '2XORE' promoter instead of Pho4 endogenous promoter
First we cloned 2XORE into Hind-Bam E.R. site of p2UG25 plasmid that has natMX4 resistence, so we create a 2XORE-natMX4 cassette,that after we can integrate in the Pho4's ORF.
2XORE Fv Hind CGCGGATCCATCAGTATTACCCTCGACCTC 2XORE Rv Bam CCCAAGCTTTGTTTGCGTGTCTATAGAAG
2XOREint Fv TCTAGATCGTCCACAAAACCGTGTATTCCCTCAGAAGTTGTACGGCCCATTGTTTGCGTGTCTATAGAAG 2XOREint Rv CAGAAGAGAAGAGATGAGCAAAGGAGACAGAACAAGAGTAGCAGAAAGTCATCGATGAATTCGAGCTCG
