FASTM (Nucleotide Databases)
Introduction
FASTM --- compare peptides to a protein sequence database.
- FASTA Website
- Download Software
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
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
