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Then the MSA was manually refined, taking into account the structural superimposition of the templates.
The resulting alignment was manually refined taking into account the predicted secondary structure of CAG and the description of the secondary structure of the crystal.
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Automatic alignments were then manually refined using Bioedit 7.0.9.0 [ 58], taking into consideration only the conserved regions of the proteins for further analyses.
Each trans-axial region-of-interest (ROI) was visually reviewed and manually refined if necessary.
Alignments were manually refined using phylogenetically conserved secondary structures.
Nucleotide sequences were aligned with Clustal W version 1.4 [39] and manually refined using MacClade v4.05 software.
Amino acid sequences were aligned with ClustalX (version 1.83) [42] and manually refined with GeneDoc (version 2.6) [43].
Alignments were performed using ClustalX [56] with default parameter values, and manually refined in GeneDoc where necessary.
All the alignments were edited and manually refined using the ED program from the MUST package [47].
The final alignment has been manually refined at the variable N-terminus and used in the subsequent analysis.
Models were manually refined with ViTO [23] using the sequence alignment of AmVKR, AgVKR, TcVKR, SpVKR and SmVKR.
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