Questions about pIRES-hrGFP1a - (Jul/10/2011 )

Hi every body,

Plasmid pIRES-hrGFP1a is a product of Stratagen. My link

In this vector there is a MCS for inserting the gene before a FLAg tag.

In the vector manual you will see this sentence: The presence of stop codons in-frame with the 3× FLAG tag must be considered when inserting genes into the MCS. it says you have to put your insert which has the stop codon before 3× FLAG tag. If so, translation will be terminated before Flag tag, how the FLAG tag peptide can be fused by our interested protein as there is a stop codon before the FLAG tag?

I need the squence of this vector but the company provides the map, not the sequence. Does anyone have the sequence information of vector pIRES-hrFGP-1a?

Are you sure that's what they mean? It sounds like just to remind you there is a stop codon at SrfI site, this needs to be removed during the cloning.