Table 1.

Sequence of ALKBH2 clones identified following methylating agent selection

Clone_selection	Non-synonimous Coding	Ori			Downstream of ori
1_MMS	G50D G64D	T113C	G189A	C409T
2_MMS	H144R	T202A	T589C		G646T	C754T
3_MMS	H144Y	C238T			C643T
4_MMS	A53T P55L	T170C	G325A	T479A	G755A
5_MMS	P27L P207S	G512A
6_MMS	H51L T121N	C328T	G592A		T840C
1_MNNG	A78V L138I R193S D201G P207L S237C	G338A
2_MNNG	P180L	G53A	G95T	G373A
3_MNNG	A97T	G53A	C198T	G477A	T687A
4_MNNG	H144Y N231D	G180A
5_MNNG	R58W A219D_P229L K255E	G44A	C116T	G180A	T666A, C667T	G850A
6_MNNG	R58W A118T	G258A			A696G
Total mutations	26 (22 new)	25			9
Expected without selection	10	1.1			5.1
Enrichment through positive selection (fold)	2.2 (footnote 2)	23 (footnote 2)			1.8 (footnote 3)
MMS	10	13			4
Expected without selection	5	0.55			2.55
Enrichment through positive selection (fold)	2.0	23			1.6
MNNG	16 (12 new)	12			5
Expected without selection	5	0.55			2.55
Enrichment through positive selection (fold)	3.2 (2.4 new)	22			2.0

Footnote 1:

Total bases: 795 × 12= 9546;

Expected frequency of bp substitutions (without selection) ~1.5/kb × 9.55kb =14

Expected frequency of amino acid substitutions (~30% synonymous (23)) ~ 10

Mutations present before randomization are underlined

Footnote 2

Total bases: 613 × 12= 7356;

Expected frequency of base-pair substitutions (without selection) ~0.15/kb ×7.36kb=1.1

Footnote 3:

Total bases: 280 × 12 =3360

Expected frequency of base-pair substitutions (without selection) ~1.5/kb × 3.4 kb= 5.1

For MMS and MNNG selections, comprising 6 and 6 clones respectively, each of the expected values calculated for the total needs to be divided by half.