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Alignments were manually refined using phylogenetically conserved secondary structures.
All the alignments were edited and manually refined using the ED program from the MUST package [47].
Nucleotide sequences were aligned with Clustal W version 1.4 [39] and manually refined using MacClade v4.05 software.
Ambiguously aligned regions were automatically detected and removed using the program Gblocks [31] and this selection was manually refined using the program ED.
Clusters were manually refined using K lustaviewa software.
Alignments were manually refined using BioEdit sequence editing software [ 43].
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PAT subgroup alignments were manually curated, and refined using UGENE (http://genome.unipro.ru) and Jalview [44].
The model of nsp4C was then manually built and refined using the programs O [44] and CNS [45].
The TEM8 model was further built manually with COOT [37] and refined using REFMAC [38] from the CCP4 suite [39].
The model of the N terminus (L24EVG to FIET135) of TraO/VirB9 was fitted first manually and then it was refined using the Flex-EM software (Topf et al, 2008; Pandurangan and Topf, 2012).
The model was refined using AutoBuster and manually rebuilt in COOT 0.6.2 and O 10.0.3.
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