Effect of deletions and point mutations on
ARS309 function. Lines represent the sequences present
in plasmids. Position 1 is the first adenine of the HpaI
site; position 349 is the cytidine of the EcoRI site.
Solid boxes show the 10 of 11 matches to the ACS at positions 31–41,
159–169, 161–171, and 231–241. The open box marks the position of
the 9 of 11 match at positions 62–72. The stippled boxes show the
positions of the 9 of 11 matches at positions 75–85 and 94–104. ×
indicates point mutations altering the ACS matches (see Table 1). Boxes
above the line indicate that the T-rich strand of the ACS is in the top
strand, and boxes below the line indicate the T-rich strand of the ACS
is in the lower strand. HFT: +, high-frequency transformation; −, no
high-frequency transformation. Stability: Numbers show percentage of
plasmid bearing cells in a logarithmic-phase culture under selection
(mean ± SEM). (A) Full-length wild-type
ARS309 (base pairs 1–349). (B) Deletion
mutant containing base pairs 1–227. (C) Deletion mutant
containing base pairs 1–147. (D) Deletion mutant
containing base pairs 54–349. (E) RsaI
subclone (base pairs 57–135). (F) Triple point mutation
altering all four 10 of 11 matches (see Table 1). (G)
Mutation altering the 9 of 11 match at positions 62–72 (see Table 1).
(H) Mutation altering the two 9 of 11 matches at
positions 75–85 and 94–104 (see Table 1).