ClustalW is a widely used system for aligning any number of homologous nucleotide or protein sequences. For multi-sequence alignments, ClustalW uses progressive alignment methods. In these, the most similar sequences, that is, those with the best alignment score are aligned first. Then progressively more distant groups of sequences are aligned until a global alignment is obtained. This heuristic approach is necessary because finding the global optimal solution is prohibitive in both memory and time requirements. ClustalW performs very well in practice. The algorithm starts by computing a rough distance matrix between each pair of sequences based on pairwise sequence alignment scores. These scores are computed using the pairwise alignment parameters for DNA and protein sequences. Next, the algorithm uses the neighbor-joining method with midpoint rooting to create a guide tree, which is used to generate a global alignment (alternatively, a guide tree in Newick format can be provided). The guide tree serves as a rough template for clades that tend to share insertion and deletion features. This generally provides a close-to-optimal result, especially when the data set contains sequences with varied degrees of divergence, so the guide tree is less sensitive to noise.
Higgins D., Thompson J., Gibson T. Thompson J. D., Higgins D. G., Gibson T. J.
CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice.
Nucleic Acids Res. 22:4673-4680. (1994)