From Tol2Kit
Revision as of 03:22, 23 March 2007 by Chi-bin (talk | contribs) (New page: === Construction details === p5E-CMV/SP6 was made by PCR amplification of the sCMV and SP6 promoters from pCS2+, using primers that added att sites, followed by a BP reaction. This 5' el...)
Construction details
p5E-CMV/SP6 was made by PCR amplification of the sCMV and SP6 promoters from pCS2+, using primers that added att sites, followed by a BP reaction.
This 5' element can be used to build pCS2-like constructs, either to make capped mRNA for injection, or to drive expression from the CMV promoter after DNA injection.
The insert has been completely sequenced, revealing two point mutations (T889C and G1508T) in the CMV promoter; however, DNA injection of constructs made with this clone show that the CMV appears to be functional.
Crudely annotated sequence
FASTA file with the full-length sequence as well as sequences of individual components:
p5E-CMV-SP6 sequence
Crude map
Screenshot from Sequencher showing locations of components:
