FASTS/TFASTSv3(1) General Commands Manual FASTS/TFASTSv3(1) NAME fasts3, fasts3_t - compare several short peptide sequences against a pro- tein database using a modified fasta algorithm. tfasts3, tfasts3_t - compare short pepides against a translated DNA data- base. DESCRIPTION fasts3 and tfasts3 are designed to compare set of (presumably non-contigu- ous) peptides to a protein (fasts3) or translated DNA (tfasts3) database. fasts3/tfasts3 are designed particularly for short peptide data from mass- spec analysis of protein digests. Unlike the traditional fasta3 search, which uses a protein or DNA sequence, fasts3 and tfasts3 work with a query sequence of the form: >tests from mgstm1 MLLE, MILGYW, MGADP, MLCYNP This sequence indicates that four peptides are to be used. When this sequence is compared against mgstm1.aa (included with the distribution), the result is: testf MILGYW----------MLLE------------MGDAP----------- :::::: :::: ::::: GT8.7 MPMILGYWNVRGLTHPIRMLLEYTDSSYDEKRYTMGDAPDFDRSQWLNEK 10 20 30 40 50 testf -------------------------------------------------- GT8.7 FKLGLDFPNLPYLIDGSHKITQSNAILRYLARKHHLDGETEEERIRADIV 60 70 80 90 100 20 testf ------------MLCYNP :::::: GT8.7 ENQVMDTRMQLIMLCYNPDFEKQKPEFLKTIPEKMKLYSEFLGKRPWFAG 110 120 130 140 150 Options fasts3 and tfasts3 can accept a query sequence from the unix "stdin" data stream. This makes it much easier to use fasta3 and its relatives as part of a WWW page. To indicate that stdin is to be used, use "-" or "@" as the query sequence file name. -b # number of best scores to show (must be < -E cutoff) -d # number of best alignments to show ( must be < -E cutoff) -D turn on debugging mode. Enables checks on sequence alphabet that cause problems with tfastx3, tfasty3, tfasta3. -E # Expectation value limit for displaying scores and alignments. Ex- pectation values for fasts3 and tfasts3 are not as accurate as those for the other fasta3 programs. -H turn off histogram display -i compare against only the reverse complement of the library sequence. -L report long sequence description in alignments -m 0,1,2,3,4,5,6,9,10 alignment display options -N # break long library sequences into blocks of # residues. Useful for bacterial genomes, which have only one sequence entry. -N 2000 works well for well for bacterial genomes. -O file send output to file -q/-Q quiet option; do not prompt for input -R file save all scores to statistics file -S # offset substitution matrix values by a constant # -s name specify substitution matrix. BLOSUM50 is used by default; PAM250, PAM120, and BLOSUM62 can be specified by setting -s P120, P250, or BL62. With this version, many more scoring matrices are available, including BLOSUM80 (BL80), and MDM_10, MDM_20, MDM_40 (M10, M20, M40). Alternatively, BLASTP1.4 format scoring matrix files can be specified. -T # (threaded, parallel only) number of threads or workers to use (set by default to 4 at compile time). -t # Translation table - tfasts3 can use the BLAST tranlation tables. See http://www.ncbi.nih.gov/htbin-post/Taxonomy/wprintgc?mode=c/. -w # line width for similarity score, sequence alignment, output. -x "#,#" offsets query, library sequence for numbering alignments -z # Specify statistical calculation. Default is -z 1, which uses regres- sion against the length of the library sequence. -z 0 disables sta- tistics. -z 2 uses the ln() length correction. -z 3 uses Altschul and Gish's statistical estimates for specific protein BLOSUM scoring matrices and gap penalties. -z 4: an alternate regression method. -Z db_size Set the apparent database size used for expectation value calcula- tions. -3 (TFASTS3 only) use only forward frame translations Environment variables: FASTLIBS location of library choice file (-l FASTLIBS) SMATRIX default scoring matrix (-s SMATRIX) SRCH_URL the format string used to define the option to re-search the data- base. REF_URL the format string used to define the option to lookup the library sequence in entrez, or some other database. AUTHOR Bill Pearson wrp@virginia.EDU local FASTS/TFASTSv3(1)
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