Primers Design For Yeast Integration

From 2007.igem.org

(Difference between revisions)
Line 3: Line 3:
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