TABLE 5.

Degenerate primers used for constructing the library comprising combinations of beneficial mutations

Primer	Oligonucleotide sequences (5′−3′) a , b
pET-ChPPK-F	CATGATTATCTGTGGCGTCATTATGTG
pET-ChPPK-R	TGCATCCATTGCCTGAAAAACAATCAG
PPK-219–346-F	TCAGGCAATGGATGCAG SAGGTAAAGATGGTA
PPK-219–346-R1	ACAGATAATCATGCT YCAGTTCAWWTTTASACGGAAC
PPK-219–346-R2	ACAGATAATCATGTG MCAGTTCAWWTTTASACGGAAC

^a Bold-marked bases in the primers indicate positions that contain the degenerate codon.

^b S = G, C (equimolar amounts); Y = C, T (equimolar amounts); W = A, T; M = A, C (equimolar amounts).