Removing CMV promoter from pcdna3.1 - Need ideas for the same (Oct/09/2006 )

Hi,
i am doing a TA cloning with insert into pcdna 3.1 V5/hisTOPO. my insert already contains a promoter and reporter gene of my interest.
i.e
CMV promotor>'my insert(promoter+reporter gene)'>V5-tag>resistance gene

my first question is can CMV promotor influence the expression of my insert?
my second question gas anyone tried to remove the CMV promotor sequence from pcdna3.1

I have tried looking for rest. sites to cleave them off but could not find suitable sites without affecting other regions of the vector.
could anyone has some idea to inactivate the CMV promoter.
thx
Raj

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i think it should b possible to remove the CMV from the vector. use bgl2 and bamh1.

Not sure how CMV may/may not influence expression of insert.

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yes its possible, but my insert also has these sites so i cant use them...i need the tags too so i like to stick with pcDNA3.1 v5 topo.
i hesitate to cleave th vector with AatII, which has 5 sites in the CMV promoter region, but the TATAA and CAAAT boxes will be still intact after religation. i am not sure that these will still drive the promoter?
any other ideas?

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y dont u put ur promoter + transgene into a different vector like pGEM or pBS or any other vector that u have.

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