Is my cDNA clone sense or antisense with respect to the gene?

cDNA is made from RNA derived from a tissue of interest. The cDNA is double-stranded (thus containing sense and antisense strands with respect to the gene of interest). Each clone is inserted into a specific vector. Orientation can be determined by looking at the cloning sites listed in genbank (or the details of our clone query). Genbank lists sequences from 5' to 3'. An example is below: Clone 4504850 (BC014748) is cloned into pCMVSPORT6 at the SalI and NotI sites of the vector. The 5' end of the insert sits at the SalI site and the 3' end sits at the NotI site. By looking at the vector map, you can see that the CMV promoter in the vector is closest to the SalI site. The promoter should then theoretically read through the insert from 5'-3' end. If the complete protein coding sequence is present in the insert, the clone should theoretically express this protein in mammalian cells. (pCMVSPORT6 is a mammalian expression vector). The CMV promoter reads through the sense strand. **NOTE: We do not test our clones for expression so we do not guarantee protein expression.

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