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================================================================== FSA Distance-based multiple sequence alignment ================================================================== FSA is a probabilistic multiple sequence alignment algorithm which uses a "distance-based" approach to aligning homologous protein, RNA or DNA sequences. Much as distance-based phylogenetic reconstruction methods like Neighbor-Joining build a phylogeny using only pairwise divergence estimates, FSA builds a multiple alignment using only pairwise estimations of homology. This is made possible by the sequence annealing technique for constructing a multiple alignment from pairwise comparisons, developed by Ariel Schwartz in "Posterior Decoding Methods for Optimization and Control of Multiple Alignments." Ph.D. Thesis, UC Berkeley. 2007. FSA brings the high accuracies previously available only for small-scale analyses of proteins or RNAs to large-scale problems such as aligning thousands of sequences or megabase-long sequences. FSA introduces several novel methods for constructing better alignments: * FSA uses machine-learning techniques to estimate gap and substitution parameters on the fly for each set of input sequences. This "query-specific learning" alignment method makes FSA very robust: it can produce superior alignments of sets of homologous sequences which are subject to very different evolutionary constraints. * FSA is capable of aligning hundreds or even thousands of sequences using a randomized inference algorithm to reduce the computational cost of multiple alignment. This randomized inference can be over ten times faster than a direct approach with little loss of accuracy. * FSA can quickly align very long sequences using the "anchor annealing" technique for resolving anchors and projecting them with transitive anchoring. It then stitches together the alignment between the anchors using the methods described above. * The included GUI, MAD (Multiple Alignment Display), can display the intermediate alignments produced by FSA, where each character is colored according to the probability that it is correctly aligned. FSA was created by Robert Bradley. It was developed by Robert Bradley, Colin Dewey, Jaeyoung Do, Sudeep Juvekar, Lior Pachter, Adam Roberts, and Michael Smoot, along with assistance from many other people. All have made intellectual contributions and contributed code. Please contact us at fsa@math.berkeley.edu with any questions, feedback, etc. If you use FSA, please cite: Bradley RK, Roberts A, Smoot M, Juvekar S, Do J, Dewey C, Holmes I, Pachter L (2009) Fast Statistical Alignment. PLoS Computational Biology. 5:e1000392. (This manuscript can be found in the doc/ directory.) ================================================================== INSTALLATION FSA is built and installed by running the following commands: ./configure make make install If you wish to install the fsa files in a location other than your system's standard directories (which usually requires root permissions), specify the top-level installation directory with the --prefix option to configure. For example, ./configure --prefix=$HOME specifies that binaries should be installed in $HOME/bin, libraries in $HOME/lib, etc.. If you wish to align long sequences, then you must install MUMmer, which FSA calls to get candidate anchors between sequences. You can download MUMmer from http://mummer.sourceforge.net/. When running configure, you must either have the mummer executable in your path or specify the executable with the --with-mummer option to configure, e.g. ./configure --with-mummer=/usr/local/MUMmer3.20/mummer if it is located at /usr/local/MUMmer3.20/mummer. FSA can also call exonerate to detect remote homology. You can download exonerate from http://www.ebi.ac.uk/~guy/exonerate/. When running configure, you must either have the exonerate executable in your path or specify the executable with the --with-exonerate option to configure, e.g. ./configure --with-exonerate=/usr/local/bin/exonerate if it is located at /usr/local/bin/exonerate. ================================================================== USAGE FSA accepts FASTA-format input files and outputs multi-FASTA alignments by default. The most basic usage is: fsa <mysequences.fa> >myalignedsequences.mfa or fsa --stockholm <mysequences.fa> >myalignedsequences.stk Many options are provided. Please see html/FAQ.html for more information. ================================================================== REFERENCES Source code in 'seq/' and 'util/' is from Ian Holmes's DART library, which is used for input and output routines. FSA's DP code was generated by HMMoC by Gerton Lunter. The 'aligner' example distributed with HMMoC, which implements a learning procedure for gap parameters, was an inspiration for FSA's learning strategies. FSA's banding code is taken directly from the 'aligner' example. The sequence annealing technique for constructing a multiple alignment from pairwise comparisons was developed by Ariel Schwartz. The implementation of sequence annealing in FSA is based on the original implementation in AMAP by Ariel Schwartz and Lior Pachter. The anchor annealing approach used in FSA is modeled after the recursive anchoring strategy used in MAVID by Nicolas Bray and Lior Pachter. The MAD GUI interface to FSA was written by Adam Roberts based on a preliminary version by Michael Smoot. Please see: I. Holmes and R. Durbin. "Dynamic Programming Alignment Accuracy." Journal of Computational Biology. 1998, 5 (3):493-504. G.A. Lunter. "HMMoC - a Compiler for Hidden Markov Models." Bioinformatics. 2007, 23 (18):2485-2487. A.S. Schwartz. "Posterior Decoding Methods for Optimization and Control of Multiple Alignments." Ph.D. Thesis, UC Berkeley. 2007. A.S. Schwartz and L. Pachter. "Multiple Alignment by Sequence Annealing." Bioinformatics. 2007, 23 (2):e24-e29. N. Bray and L. Pachter "MAVID: Constrained Ancestral Alignment of Multiple Sequences." Genome Research. 2004, 14:693-699. ================================================================== COPYRIGHT Copyright (C) 2008. The Regents of the University of California (Regents). All Rights Reserved. Permission to use, copy, modify, and distribute this software and its documentation for educational, research, and not-for-profit purposes, without fee and without a signed licensing agreement, is hereby granted, provided that the above copyright notice, this paragraph and the following two paragraphs appear in all copies, modifications, and distributions. Contact The Office of Technology Licensing, UC Berkeley, 2150 Shattuck Avenue, Suite 510, Berkeley, CA 94720-1620, (510) 643-7201 for commercial licensing opportunities. Created by Robert K. Bradley, Departments of Math and Molecular and Cell Biology, University of California, Berkeley. IN NO EVENT SHALL REGENTS BE LIABLE TO ANY PARTY FOR DIRECT, INDIRECT, SPECIAL, INCIDENTAL, OR CONSEQUENTIAL DAMAGES, INCLUDING LOST PROFITS, ARISING OUT OF THE USE OF THIS SOFTWARE AND ITS DOCUMENTATION, EVEN IF REGENTS HAS BEEN ADVISED OF THE POSSIBILITY OF SUCH DAMAGE. REGENTS SPECIFICALLY DISCLAIMS ANY WARRANTIES, INCLUDING, BUT NOT LIMITED TO, THE IMPLIED WARRANTIES OF MERCHANTABILITY AND FITNESS FOR A PARTICULAR PURPOSE. THE SOFTWARE AND ACCOMPANYING DOCUMENTATION, IF ANY, PROVIDED HEREUNDER IS PROVIDED "AS IS". REGENTS HAS NO OBLIGATION TO PROVIDE MAINTENANCE, SUPPORT, UPDATES, ENHANCEMENTS, OR MODIFICATIONS. ================================================================== LICENSE Please see the file LICENSE distributed with FSA. FSA is free software; you can redistribute it and/or modify it under the terms of the GNU General Public License as published by the Free Software Foundation; either version 3 of the License, or (at your option) any later version. FSA is distributed in the hope that it will be useful, but WITHOUT ANY WARRANTY; without even the implied warranty of MERCHANTABILITY or FITNESS FOR A PARTICULAR PURPOSE. See the GNU General Public License for more details. You should have received a copy of the GNU General Public License along with FSA; if not, write to the Free Software Foundation, Inc., 51 Franklin St, Fifth Floor, Boston, MA 02110-1301 USA ================================================================== VERSION HISTORY 1.01, 7/10/2008 -- Initial import to Git (Robert Bradley) 1.02, 7/21/08 -- Faster anchor annealing (Robert Bradley) -- GUI added to package (Adam Roberts) 1.03, 7/25/08 -- Improved GUI features (Adam Roberts) 1.04, 7/28/08 -- Moved to autotools (Colin Dewey) 1.05, 8/21/08 -- Optional anchor annealing in protein space (Robert Bradley) -- Reduced GUI memory usage (Adam Roberts) -- Many accuracy measures for GUI (Adam Roberts and Robert Bradley) 1.06, 8/29/08 -- Homology no longer forced at anchor boundaries (Robert Bradley) -- Scripts for evaluating alignment accuracies according to FSA's model (Robert Bradley) 1.07, 9/2/08 -- GUI bugfix for displaying extremely certain alignments (Robert Bradley) (Alignment inference is unaffected.) 1.08, 9/15/08 -- Miscellaneous compilation fixes (Robert Bradley) -- Fix for learning on highly-conserved alignments (Robert Bradley) 1.09, 10/22/08 -- Much faster sequence annealing (Robert Bradley, Colin Dewey & Lior Pachter) -- Better memory usage for long sequences (Robert Bradley) -- Optional Mercator constraints (Robert Bradley and Colin Dewey) -- Optional anchoring with exonerate (Robert Bradley) -- GUI can export TIFF images (Robert Bradley) -- gapcleaner fix (Robert Bradley) -- Parallelization mode (Jaeyoung Do and Colin Dewey) -- Database mode (Jaeyoung Do and Colin Dewey) -- Added submitted manuscript as manual. (The below versions are labeled as "major.minor.bugfix") 1.10.0, 10/30/08 -- --nucprot option and utility programs translate and prot2codon (Robert Bradley & Colin Dewey) -- Bugfixes for anchoring with both Mercator & exonerate (Robert Bradley & Colin Dewey) -- Bugfixes for parallelization & database modes (Jaeyoung Do & Colin Dewey) 1.11.0, 11/11/08 -- housekeeping tasks (Robert Bradley): - gaps stripped from input sequences - error thrown if 2 alignable sequence are left unaligned due to RAM constraints -- hardmasked sequence is left unaligned by default (Robert Bradley) 1.11.1, 11/13/08 -- Bugfixes for anchoring with both Mercator & exonerate (Robert Bradley & Colin Dewey) -- Compilation fixes for g++ 4.4 (Robert Bradley) 1.11.2, 11/14/08 -- Bugfix for GUI compilation (Colin Dewey) 1.12.0, 11/22/08 -- Perl scripts to interact with FSA-Mercator whole-genome alignments (Robert Bradley & Colin Dewey) -- Switch to using only ungapped exonerate anchors (Robert Bradley & Colin Dewey) 1.12.1, 11/27/08 -- Source code documentation updated for Doxygen compatibility (Robert Bradley) 1.13.0, 1/9/09 -- C++ tools for working with FSA-Mercator whole-genome alignments (Robert Bradley & Colin Dewey) (Note that the deprecated Perl scripts had a serious bug for - strand extraction!) -- Bugfix for softmasking with exonerate (Robert Bradley & Colin Dewey) -- Completely re-written sequence and alignment representation code (Robert Bradley) -- Use a model with 2 sets, rather than 1 set, of indel states by default -- Turn iterative refinement on by default with an unlimited number of steps 1.13.1, 1/9/09 -- Bugfix and better automated options for --nucprot (Robert Bradley) 1.13.2, 1/12/09 -- Compilation fixes for g++ 4.4 (Robert Bradley) 1.13.3, 1/14/09 -- Bugfix for alphabet case-sensitivity during learning (Robert Bradley) 1.13.4, 2/6/09 -- Bugfix for % ID calculation for sliced alignments (Robert Bradley) -- More intuitive default anchoring options (Colin Dewey) -- Bugfix for hardmasked error after compilation without MUMmer on OS X 10.4 (Robert Bradley) 1.14.0, 3/11/09 -- Added experimental --tree-weights option (Robert Bradley) -- Added experimental --load-probs option (Robert Bradley) -- Changed default behavior when two sequences have no detectable homology; see new option --require-homology (Robert Bradley) -- New utility program slice_fasta to extract subsequences from FASTA files (Robert Bradley). 1.14.1, 3/20/09 -- Changed Mercator-related short options to use upper-case letters (Robert Bradley). -- Mercator slicing stuff now assumes a + strand if strand is unspecified; previously implicitly assumed - (Robert Bradley) -- Tiny bugfix for map_gff_coords: --force-entry was accidentally on by default (Robert Bradley) -- Bugfix for GUI compilation on Linux (Colin Dewey) 1.14.2, 3/23/09 -- Updated FSA preprint (Robert Bradley). 1.14.3, 3/31/09 -- Bugfix for recognizing DNA sequences with a lot of hardmasked sequence as protein (Robert Bradley) 1.14.4, 4/2/09 -- Compilation bugfixes for some Linux systems where floor isn't declared (Robert Bradley) -- Added FSA preprint accepted by PLoS Computational Biology (Robert Bradley) 1.14.5, 4/14/09 -- Handle "NA" fields in Mercator maps (Robert Bradley) -- Bugfix for automatic available RAM detection for some Linux distributions (Colin Dewey) -- Parallelization and database modes are disabled by default (Colin Dewey) -- Bugfix for strange behavior where all sequences are left unaligned under some older compilers (Colin Dewey) 1.15.0, 6/12/09 -- Options to use k-mer similarities to choose sequence pairs for alignment, rather than just randomly (Robert Bradley & Colin Dewey) * New options include --mst-min, --mst-max, --mst-palm, --degree and --kmer. * Deprecated --training-number and --training-fraction options. -- Memory usage in --maxsn mode reduced by ~2X (Robert Bradley & Colin Dewey) The above two changes result in significantly-better large (> 100 sequences) alignments. -- New program slice_fasta_gff (Robert Bradley) -- Compiler flags for 64-bit systems to ensure that all available RAM is used if requested (Robert Bradley) -- Published manuscript (http://www.ploscompbiol.org/article/info:doi/10.1371/journal.pcbi.1000392) included in doc/ -- Bugfix for automatic setting of --maxram for available RAM assessment (Colin Dewey) 1.15.1, 7/15/09 -- Refactored alignment code to speed up the FSA-Mercator tools (Robert Bradley) 1.15.2, 7/16/09 -- Minor compilation fix for g++ 4.1.2 (Robert Bradley) 1.15.3, 10/14/09 -- Minor bugfix for running in --anchored --translated mode (Robert Bradley) 1.15.4, 8/5/09 -- Minor bugfix for Mercator constraints which are all Ns in one species (Robert Bradley) 1.15.5, 8/10/09 -- Use tr1/unordered_set for annealing if available (Robert Bradley) 1.15.6, 6/22/11 -- Minor compilation fix for gcc 4.5.2 (Robert Bradley) 1.15.7, 1/30/12 -- Minor compilation fix for gcc 4.6 (Robert Bradley)
Source: README, updated 2012-01-30