Primers Design For Yeast Integration
From 2007.igem.org
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2XORE Fv Hind CGCGGATCCATCAGTATTACCCTCGACCTC | 2XORE Fv Hind CGCGGATCCATCAGTATTACCCTCGACCTC | ||
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2XORE Rv Bam CCCAAGCTTTGTTTGCGTGTCTATAGAAG | 2XORE Rv Bam CCCAAGCTTTGTTTGCGTGTCTATAGAAG | ||
+ | |||
2XOREint Fv TCTAGATCGTCCACAAAACCGTGTATTCCCTCAGAAGTTGTACGGCCCATTGTTTGCGTGTCTATAGAAG | 2XOREint Fv TCTAGATCGTCCACAAAACCGTGTATTCCCTCAGAAGTTGTACGGCCCATTGTTTGCGTGTCTATAGAAG | ||
+ | |||
2XOREint Rv CAGAAGAGAAGAGATGAGCAAAGGAGACAGAACAAGAGTAGCAGAAAGTCATCGATGAATTCGAGCTCG | 2XOREint Rv CAGAAGAGAAGAGATGAGCAAAGGAGACAGAACAAGAGTAGCAGAAAGTCATCGATGAATTCGAGCTCG |
Revision as of 18:27, 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