Adding T7 promoters

[Rich Dudley]

We want to add a T7 promoter to a fusion vector which does not contain a
T7 promoter, so that we may make RNA.  The RNA would then be injected
into Xenopus oocytes.

There are 2 restriction sites, between which we could ligate a linker
containing a consensus T7 promoter.  Since we have some leeway in the
exact placement of the promoter, is there any particular advantage in
having the first transcribed base in frame with the ATG?  The ATG will
be the 5th codon from the first transcribed base in the scheme I am
planning to use; unfortunately, I can't make the start codon the first
transcribed base (but I don't expect a little 5' UTR to be a problem).

Thanks!
rich

--- --- --- -- -- -- --- --- ---
Richard J. Dudley (rdudley+ at pitt.edu)
Research Specialist V
Dept. of Cell Biology and Physiology
University of Pittsburgh
http://www.cbp.pitt.edu
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