Construction of synthetic DNA arrays for RMCE cassettes. (A) Repeat DNA elements were amplified by rolling circle amplification (RCA). Such amplification generates RCA products of 2–5 kb in size. (B) RCA products were assembled into long arrays in pYB using in vivo recombination in yeast. Repeat-specific hooks in pYB were released by MluI digestion removing the pUC linker. RCA-amplified repeats and linearized vector were cotransformed into VL6-48 yeast cells. End-to-end homologous recombination of RCA products followed by interaction of the recombined fragments with the vector allows recovery of large repeat arrays in yeast. (C) Synthetic arrays used for analysis.
