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Local conservation was avoided by dividing the alignment length of each aligned sequence by its total length.

A stepwise approach allows progressive addition of each aligned cluster to the seed alignment.

First, an alignment score is calculated for each aligned column by a method similar to that used in the CLUSTALX package [42].

Final assembling of each aligned group was performed using profile alignment generating a final dataset of 6,244 sites.

This approach formally considers the relative evolutionary distance between the member genomes and yields a single value for each aligned position in a multiple sequence alignment.

For each aligned base of the consensus sequence, the alignment algorithm generates [49] a quality score (pSNP).

A large set of random alignments was simulated using all curated domains, each aligned with 100 domains from different superfamilies.

Each aligned read was then consolidated into a single genomic alignment, including intronic splice gaps.

In the given multiple motif alignment, we computed the motif conservation score (MCS) at each aligned position by taking the average of the aligned affinity scores.

Genes common to all three C. raciborskii strains were each aligned as orthologous clusters using Muscle [ 47] and the alignments assessed for synonymous, non-synonymous and indel mutations.

Each aligned sequence read is considered one hit at the 5′ end of its alignment to the reference assembly.

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