FASTM (Nucleotide Databases)

Introduction

FASTM --- compare peptides to a protein sequence database.

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How to use this tool

Running a tool from the web form is a simple multiple steps process, starting at the top of the page and following the steps to the bottom.

Each tool has at least 2 steps, but most of them have more:

  • The first steps are usually where the user sets the tool input (e.g. sequences, databases...)
  • In the following steps, the user has the possibility to change the default tool parameters
  • And finally, the last step is always the tool submission step, where the user can specify a title to be associated with the results and an email address for email notification. Using the submit button will effectively submit the information specified previously in the form to launch the tool on the server

Note that the parameters are validated prior to launching the tool on the server and in the event of a missing or wrong combination of parameters, the user will be notified directly in the form.

Step 1 - Database

Databases

The databases to run the sequence similarity search against. Multiple databases can be used at the same time

Database Name Description Abbreviation
EMBL-Bank EMBL-Bank constitutes Europe's primary nucleotide sequence resource. Main sources for DNA and RNA sequences are direct submissions from individual researchers, genome sequencing projects and patent applications.
EMBL Release The quarterly release of the EMBL nucleotide sequence databank (EMBL-Bank) em_rel
EMBL Environmental Sequences from Environmental Samples. em_rel_env
EMBL EST Environmental em_rel_est_env
EMBL GSS Environmental em_rel_gss_env
EMBL HTC Environmental em_rel_htc_env
EMBL HTG Environmental em_rel_htg_env
EMBL Patent Environmental em_rel_pat_env
EMBL Standard Environmental em_rel_std_env
EMBL TSA Environmental em_rel_tsa_env
EMBL Fungi Sequences from Fungi. em_rel_fun
EMBL EST Fungi em_rel_est_fun
EMBL GSS Fungi em_rel_gss_fun
EMBL HTC Fungi em_rel_htc_fun
EMBL HTG Fungi em_rel_htg_fun
EMBL Patent Fungi em_rel_pat_fun
EMBL Standard Fungi em_rel_std_fun
EMBL STS Fungi em_rel_sts_fun
EMBL TSA Fungi em_rel_tsa_fun
EMBL Human Sequences from Human. em_rel_hum
EMBL EST Human em_rel_est_hum
EMBL GSS Human em_rel_gss_hum
EMBL HTC Human em_rel_htc_hum
EMBL HTG Human em_rel_htg_hum
EMBL Patent Human em_rel_pat_hum
EMBL Standard Human em_rel_std_hum
EMBL STS Human em_rel_sts_hum
EMBL Invertebrate Sequences from invertebrate organisms. em_rel_inv
EMBL EST Invertebrate em_rel_est_inv
EMBL GSS Invertebrate em_rel_gss_inv
EMBL HTC Invertebrate em_rel_htc_inv
EMBL HTG Invertebrate em_rel_htg_inv
EMBL Patent Invertebrate em_rel_pat_inv
EMBL Standard Invertebrate em_rel_std_inv
EMBL STS Invertebrate em_rel_sts_inv
EMBL TSA Invertebrate em_rel_tsa_inv
EMBL Mammal Sequences from mammals, excluding rodents and humans. em_rel_mam
EMBL EST Mammal em_rel_est_mam
EMBL GSS Mammal em_rel_gss_mam
EMBL HTC Mammal em_rel_htc_mam
EMBL HTG Mammal em_rel_htg_mam
EMBL Patent Mammal em_rel_pat_mam
EMBL Standard Mammal em_rel_std_mam
EMBL STS Mammal em_rel_sts_mam
EMBL TSA Mammal em_rel_tsa_mam
EMBL Mouse Sequences from mus musculus. em_rel_mus
EMBL EST Mouse em_rel_est_mus
EMBL GSS Mouse em_rel_gss_mus
EMBL HTC Mouse em_rel_htc_mus
EMBL HTG Mouse em_rel_htg_mus
EMBL Patent Mouse em_rel_pat_mus
EMBL Standard Mouse em_rel_std_mus
EMBL STS Mouse em_rel_sts_mus
EMBL Phage Sequences from bacteria phages. em_rel_phg
EMBL GSS Phage em_rel_gss_phg
EMBL HTG Phage em_rel_htg_phg
EMBL Patent Phage em_rel_pat_phg
EMBL Standard Phage em_rel_std_phg
EMBL Plant Sequences from plants. em_rel_pln
EMBL EST Plant em_rel_est_pln
EMBL GSS Plant em_rel_gss_pln
EMBL HTC Plant em_rel_htc_pln
EMBL HTG Plant em_rel_htg_pln
EMBL Patent Plant em_rel_pat_pln
EMBL Standard Plant em_rel_std_pln
EMBL STS Plant em_rel_sts_pln
EMBL TSA Plant em_rel_tsa_pln
EMBL Prokaryote Sequences from prokaryotes. em_rel_pro
EMBL EST Prokaryote em_rel_est_pro
EMBL GSS Prokaryote em_rel_gss_pro
EMBL HTC Prokaryote em_rel_htc_pro
EMBL HTG Prokaryote em_rel_htg_pro
EMBL Patent Prokaryote em_rel_pat_pro
EMBL Standard Prokaryote em_rel_std_pro
EMBL STS Prokaryote em_rel_sts_pro
EMBL Rodent Sequences from rodents, but not mouse. em_rel_rod
EMBL EST Rodent em_rel_est_rod
EMBL GSS Rodent em_rel_gss_rod
EMBL HTC Rodent em_rel_htc_rod
EMBL HTG Rodent em_rel_htg_rod
EMBL Patent Rodent em_rel_pat_rod
EMBL Standard Rodent em_rel_std_rod
EMBL STS Rodent em_rel_sts_rod
EMBL TSA Rodent em_rel_tsa_rod
EMBL Synthetic Sequences from synthetic constructs. em_rel_syn
EMBL Patent Synthetic em_rel_pat_syn
EMBL Standard Synthetic em_rel_std_syn
EMBL Transgenic Sequences from transgenic contructs. em_rel_tgn
EMBL Standard Transgenic em_rel_std_tgn
EMBL GSS Transgenic em_rel_gss_tgn
EMBL Unclassified Sequences from unspecified origin. em_rel_unc
EMBL EST Unclassified em_rel_est_unc
EMBL Patent Unclassified em_rel_pat_unc
EMBL Standard Unclassified em_rel_std_unc
EMBL Viral Sequences from Viruses. em_rel_vrl
EMBL EST Viral em_rel_est_vrl
EMBL GSS Viral em_rel_gss_vrl
EMBL HTG Viral em_rel_htg_vrl
EMBL Patent Viral em_rel_pat_vrl
EMBL Standard Viral em_rel_std_vrl
EMBL TSA Viral em_rel_tsa_vrl
EMBL Vertebrate Sequences from vertebrates, excluding human, mouse and rodents. em_rel_vrt
EMBL EST Vertebrate em_rel_est_vrt
EMBL GSS Vertebrate em_rel_gss_vrt
EMBL HTC Vertebrate em_rel_htc_vrt
EMBL HTG Vertebrate em_rel_htg_vrt
EMBL Patent Vertebrate em_rel_pat_vrt
EMBL Standard Vertebrate em_rel_std_vrt
EMBL STS Vertebrate em_rel_sts_vrt
EMBL TSA Vertebrate em_rel_tsa_vrt
EMBL Updates Daily updates to the quarterly EMBL nucleotide sequence databank (EMBL-Bank) release emnew
EMBL Coding Sequence The nucleotide sequences of the coding sequence (CDS) features in the EMBL nucleotide sequence databank (EMBL-Bank). emblcds
Others
EMBL Expressed Sequence Tag em_rel_est
EMBL Genome Survey Sequence em_rel_gss
EMBL High Throughput cDNA em_rel_htc
EMBL High Throughput Genome em_rel_htg
EMBL Patent em_rel_pat
EMBL Standard em_rel_std
EMBL Sequence Tagged Site em_rel_sts
EMBL Transcriptome Shotgun Assembly em_rel_tsa
EMBL Release and Updates emall
EMBL Vectors Sequencing vectors extracted from the EMBL nucleotide sequence databank (EMBL-Bank) emvec
IMGT
IMGT/LIGM-DB The immunoglobulins and T cell receptors (LIGM-DB) section of the international immunogenetics (IMGT) database imgtligm
IMGT/HLA The human major histocompatibility complex (HLA) section of the the international immunogenetics (IMGT) database imgthla
IPD-KIR Human Killer-cell Immunoglobulin-like Receptors (KIR) sequence in the Immuno Polymorphism Database (IPD) ipdkir
IPD-MHC Major Histocompatibility Complex (MHC) section of the Immuno Polymorphism Database (IPD) ipdmhc
Patents
NR Patent DNAs Level-1 Non-redundant Patent DNAs sequences Level 1 covering patent data from the EMBL-Bank nrnl1
NR Patent DNAs Level-2 Non-redundant Patent DNAs sequences Level 2 covering patent data from the EMBL-Bank nrnl2
Structure
Nucleotide Structure Sequences Nucleotide sequences from structures described in the Brookhaven Protein Data Bank(PDB) pdbna

Default value is: EMBL TSA Vertebrate [em_rel_tsa_vrt]

Step 2 - Sequence

Sequence Input Window

The input set of peptide or nucelotide sequence fragments are described using a modified fasta sequence format. This comprises a fasta header line with an identifier for the set of sequences and optionally a description, followed by the individual sequences each starting on a newline and separated with commas. Partially formatted sequences are not accepted. Adding a return to the end of the sequence may help certain applications understand the input. Note that directly using data from word processors may yield unpredictable results as hidden/control characters may be present.

Sequence File Upload

The input file containing the set of peptide or nucelotide sequence fragments to search with uses a modified fasta sequence format. This comprises a fasta header line with an identifier for the set of sequences and optionally a description, followed by the individual sequences each starting on a newline and separated with commas. Word processors files may yield unpredictable results as hidden/control characters may be present in the files. It is best to save files with the Unix format option to avoid hidden Windows characters.

Sequence Type

Indicates if the query sequence is protein, DNA or RNA. Used to force FASTA to interpret the input sequence as specified type of sequence (via. the '-p', '-n' or '-U' options), this prevents issues when using nucleotide sequences that contain many ambiguous residues.

Type Abbreviation
DNA dna
RNA rna

Default value is: DNA [dna]

Step 3 - Parameters

Program

The FASTA program to be used for the Sequence Similarity Search

Program Name Description Abbreviation
FASTM Compare short peptides to a protein sequence database. fastm

Default value is: FASTM [fastm]

Matrix

The comparison matrix to be used to score alignments when searching the database

Matrix Name Abbreviation
N/A none
Additional information

Match/mismatch__scores

Specify match/mismatch scores for DNA comparisons. The default is "+5/-4". "+3/-2" can perform better in some cases.

Match/mismatch_scores Abbreviation
+2/-2 +2/-2
+5/-4 +5/-4
+3/-2 +3/-2
N/A none

Default value is: +2/-2

Gap Open Penalty

Score for the first residue in a gap.

Default value is: -14

Additional information

Gap Extend Penalty

Score for each additional residue in a gap.

Default value is: -4

Additional information

KTUP

FASTA uses a rapid word-based lookup strategy to speed the initial phase of the similarity search. The KTUP is used to control the sensitivity of the search. Lower values lead to more sensitive, but slower searches.

Default value is: 2

Expectation Upper Limit

Limits the number of scores and alignments reported based on the expectation value. This is the maximum number of times the match is expected to occur by chance.

Default value is: 10

Expectation Lower Limit

Limit the number of scores and alignments reported based on the expectation value. This is the minimum number of times the match is expected to occur by chance. This allows closely related matches to be excluded from the result in favor of more distant relationships.

Default value is: 0 (default) [0]

Strand

For nucleotide sequences specify the sequence strand to be used for the search. By default both upper (provided) and lower (reverse complement of provided) strands are used, for single stranded sequences searching with only the upper or lower strand may provide better results.

Value
none
both
top
bottom

Default value is: both

Histogram

Turn on/off the histogram in the FASTA result. The histogram gives a qualitative view of how well the statistical theory fits the similarity scores calculated by the program.

Default value is: no [false]

Filter

Filter regions of low sequence complexity. This can avoid issues with low complexity sequences where matches are found due to composition rather then meaningful sequence similarity. However in some cases filtering also masks regions of interest and so should be used with caution.

Value Description
none No filtering of the query sequence.
dust Uses the DUST filter (Tatusov and Lipman) to mask simple repeats in DNA/RNA sequences.

Default value is: none

Statistical Estimates

The statistical routines assume that the library contains a large sample of unrelated sequences. Options to select what method to use include regression, maximum likelihood estimates, shuffles, or combinations of these.

Name Description Value
Regress Uses a weighted regression of average score vs library sequence length. 1
MLE Uses Maximum Likelihood Estimates of Lambda and K. 2
Altshul-Gish Uses Altschul-Gish parameters (Altschul and Gish, 1996). 3
Regress/shuf. Estimate the statistical parameters from shuffled copies of each library sequence using the Regress method above. 11
MLE/shuf. Estimate the statistical parameters from shuffled copies of each library sequence using the Maximum Likelihood Estimates method above. 12

Default value is: Regress [1]

Scores

Maximum number of match score summaries reported in the result output.

Default value is: 50

Alignments

Maximum number of match alignments reported in the result output.

Default value is: 50

Sequence Range

Specify a range or section of the input sequence to use in the search. Example: Specifying '34-89' in an input sequence of total length 100, will tell FASTA to only use residues 34 to 89, inclusive.

Default value is: START-END

Database Range

Specify the sizes of the sequences in a database to search against. For example: 100-250 will search all sequences in a database with length between 100 and 250 residues, inclusive.

Default value is: START-END

HSPs

Turn on/off the display of all significant alignments between query and library sequence.

Default value is: no [false]

Score Format

Different score report formats.

Name Description Value
Default Default FASTA score format default
-m 9 -- with coordinates scores and %identity To extend scores report with coordinates scores and %identity. 9
-m 9C -- with CIGAR alignment To display an alignment code in CIGAR format. 9C
-m 9c -- with encoded alignment To extend scores report with coordinate, %identity and encoded alignment details. 9c
-m 9i -- with identity and length To extend scores report with %identity and length only. 9i

Default value is: Default [default]

Step 4 - Submission

Job title

It's possible to identify the tool result by giving it a name. This name will be associated to the results and might appear in some of the graphical representations of the results.

Email Notification

Running a tool is usually an interactive process, the results are delivered directly to the browser when they become available. Depending on the tool and its input parameters, this may take quite a long time. It's possible to be notified by email when the job is finished by simply ticking the box "Be notified by email". An email with a link to the results will be sent to the email address specified in the corresponding text box. Email notifications require valid email addresses.

Email Address

If email notification is requested, then a valid Internet email address in the form joe@example.org must be provided. This is not required when running the tool interactively (The results will be delivered to the browser window when they are ready).

References

Getting more from less: algorithms for rapid protein identification with multiple short peptide sequences.
(2002 Feb) Molecular & cellular proteomics : MCP 1 (2) :139-47
Rapid and sensitive sequence comparison with FASTP and FASTA.
(1990) Methods in enzymology 183 :63-98
Improved tools for biological sequence comparison.
(1988 Apr) Proceedings of the National Academy of Sciences of the United States of America 85 (8) :2444-8

Contact details

Support:

For Support on this service: Please contact EBI support at http://www.ebi.ac.uk/support/

The Author:

William R. Pearson (email: wrp@virginia.edu)
Department of Biochemistry
Box 440, Jordan Hall
U. of Virginia
Charlottesville, VA