TABLE 1

Sequences for one of the two oligonucleotides used in the construction of each of the CYP2A13 mutations

For each mutation, the second oligonucleotide used was a perfect complement of the oligonucleotide sequence shown. Bold indicates changes from the CYP2A13 sequence. Underline indicates the location of the desired mutation. Italics indicate changes that alter a restriction site and were used to facilitate identification of plasmids containing the desired mutation.


Mutation

Sequence (5′ to 3′)

Restriction Site Altered
L110V CC ACC TTC GAC TGG GTC TTC AAA GGC TAT GGC Delete SapI
A117V GGC TAT GGC GTG GTC TTC AGC AAC GGG Add BbsI
S208I CGC ATG ATG CTG GGA ATC TTC CAG TTC ACG GCA ACC Delete HindIII
A213S GGA AGC TTC CAG TTC ACG TCG ACC TCC ACG GGG CAG C New SalI
F300I CC CTG AAC CTC TTC ATT GCG GGT ACC GAG ACC GTG AGC ACC Add KpnI
A301G CC CTG AAC CTC TTC TTT GGG GGT ACC GAG ACC GTG AGC Add KpnI
M365V C CAA AGA TTT GGA GAC GTC CTC CCC ATG GGT TTG G Add AatII
L366I C CAA AGA TTT GGA GAC ATG ATC CCG ATG GGT TTG GCC C Delete NcoI
G369S GGA GAC ATG CTC CCC ATG AGC TTA GCC CAC AGG GTC AAC AAG G Add Bpu1102I
H372R
CCC ATG GGT TTG GCC CGC AGG GTT AAC AAG GAC ACC AAG TTT CG
Add HpaI