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Synonymous substitution rates between individual syntenic gene pairs were calculated within the SynMap utility for aligned coding sequences of gene pairs guided by the alignment of the translated coding sequences of gene pairs by nwalign (http://pypi.python.org/pypi/nwalign/).org/pypi/nwalign/
CDS sequences from each single-copy family were aligned, guided by MUSCLE alignments of protein sequences and concatenated to one super gene for each species.
Third, guided by the alignment information to the reference genome, the PE multipositional de Bruijn graph is less affected by regionally low read coverage that often gives rise to incomplete paths in the conventional de Bruijn graph.
Multiple nucleotide alignments were built guided by previous alignments of amino acid sequences using Muscle [ 24] and Pal2Nal [ 25].
Selection of the CEL6A sites for engineering was based on knowledge of the enzyme structure and guided by sequence alignments.
The COI coding DNA sequence alignments were guided by pre-aligned COI protein sequences by using MEGA3 software [4].
RNI was first aligned to Homo_7 using these methods, and adjusted manually, and then aligned to the full alignment of all human PRAMEs guided by the Homo_7 alignment.
The analysis was performed on nucleotide sequence alignments of core regions, with gaps removed, of human and three closely related vertebrate orthologs, guided by protein sequence alignments.
The divide-and-conquer strategy has also been applied to classify protein sequences based on conserved profiles, with MSA of proteins guided by multiple-profile alignments [ 30].
Since EC3 loop contains a motif similar to ion channel selectivity filter, we generated an alignment guided by presence of similar motifs to generate Model 3 (using alignment in Fig. S2C).
The sequences in each nucleotide sequence dataset were aligned manually, and alignment was guided by the corresponding computer-generated amino acid sequence alignment.
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