CAG Promoter (pEMS1277)

CAG Description:

The CAG promoter is generally reported to have a wide expression throughout cell types and development (resources). To act as a positive control and to generate new ubiquitously expressing resources, the Pleiades Promoter Project isolated the CAG promoter from the InvivoGen pDRIVE-CAG vector and introduced it into the multiple cloning site (MCS) of Pleiades expression vector pEMS1307 (EGFP reporter flanked by full frt sites) to make plasmid pEMS1277 with CAG driving EGFP. The CAG-EGFP (pEMS1277) was introduced by homologous recombination as single copy insertion into the Hprt1b-m3 locus in the X Chromosome of mouse embryonic stem cells (ESCs) mEMS1218. The resulting PCR validated four ESC lines were mEMS2077, mEMS2079, mEMS2084, and mEMS2096. Chimeras were generated using mEMS2077 this cell line were used to generate the images showing ubiquitous expression as expected of this positive control.

In a parallel experiment, also to generate a positive control, the CAG promoter was introduced into the MCS of Pleiades expression vector pEMS1157 (EGFP reporter flanked by minimal frt sites) to make plasmid pEMS1157 with CAG driving EGFP.

Note: a promoterless negative control (pEMS1306) has also been generated.

CAG Promoter Resources:

CAG-EGFP
	plasmid pEMS1277 (in backbone pEMS1307)
	ESCs mEMS2077, mEMS2079, mEMS2084, and mEMS2096
	chimeras generated using mEMS2077
	images from mEMS2077
	expression ubiquitous
Resource Files
	CAG Sequence	Jones lab
	CAG MiniPromoter Design	Wasserman lab
	CAG pEMS1277 Vector NTI file  (Requires VectorNTI Sofware)	Holt lab
	CAG pEMS1277 Vector Map	Holt lab
	CAG pEMS1277 Genotyping Assay	Simpson lab
	CAG pEMS1277 ESC description	Simpson lab
	CAG pEMS1277 Mouse description	Simpson lab

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