I would like to design a targeting vector to insert right before the stop codon of a gene with the following cassette: SA-T2A-Puro-pA-CAG-GFP-pA.
1) What sequence should I design for the splice acceptor?
2) If the splice donor is GT and the splice acceptor is AT, how does the spliceosome know this particular AT to choose for splicing?
3) Is the splice acceptor/splice donor sequence gene specific?
4) Is there a way to prevent any downstream AT from being chosen as the splice acceptor?
Edited by labstud4, 15 May 2014 - 02:55 PM.