NADPH biosensor-based identification of an alcohol dehydrogenase variant with improved catalytic properties caused by a single charge reversal at the protein surface

AMB Express

Table 2 Oligonucleotides used in this study

Oligonucleotide	Sequence (5′ → 3′) and properties
pSenSox-Lbadh⁻fw	CAATTTCACACAGGAAACAGGCGGCCGCATGTCTAACCGTTTGGATG
pSenSox-Lbadh⁻rv	CTCTCATCCGCCAAAACAGAGAATTCCTATTGAGCAGTGTAGCC
pSenSox-Lbadh^Library sequencing fw	TAATCATCGGCTCGTATAATGTGTG
pSenSox-Lbadh^Library sequencing rv	GCTTCTGCGTTCTGATTTAATCTG
Lbadh-mutagenesis A412G fw	GATGAAGATGGTTGGACCGAACTGTTTGATGCAACC
Lbadh-mutagenesis A412G rv	GGTTGCATCAAACAGTTCGGTCCAACCATCTTCATC
pASK-IBA5plus-Lbadh^K71E sequencing fw	GAAATAATTTTGTTTAACTTTAAGAAGG
pASK-IBA5plus-Lbadh^K71E sequencing rv	CCATTTTTCACTTCACAGGTCAAGC

Overlapping regions required for Gibson assembly in oligonucleotides used for cloning of the Lbadh^Library genes into pSenSox cut with EcoRI and HindIII are shown in bold