To make primers at the ends of the coding sequence, the neighbouring noncoding sequence are required. Two options are available:To give the upstream and downstream sequence separately.
Add to the following textarea the list of mutations desired, such as A13P. If more complex mutations, such as degenerate codons are to be use mark the destination codon in square brakets after the AA residue and its number, e.g. A13[NNK]. For mutations to multiple amino acids (e.g. A13GP) please specify the required codon for now. If considering degenerate codons and are not sure which to pick, try using the helper tool.