Submission of a local file
(HTML 3.0 or higher)
Submission by pasting sequences:
At most 1000 sequences, at least 50 nucleotides long, at most 10,000,000 nucleotides per submission.
For publications of results, please cite:
Two methods for improving performance of an HMM and their application
for gene finding.
In Proc. of Fifth Int. Conf. on Intelligent Systems for Molecular Biology,
ed. Gaasterland, T. et al., Menlo Park, CA: AAAI Press, 1997, pp. 179-186.
The server has two forms:
You can either submit a local file with sequences in fasta format,
or paste your sequence into the window. Then you select which organism
the sequence is from and what options you would like to use.
Optionally you can also specify known annotation from e.g. database hits.
There are currently models for vertebrate and C. elegans. The vertebrate
model is trained entirely on human genes, but it should work reasonably
well for other vertebrates.
DNA sequences must be in FASTA format which looks like this example:
>SEQ1 Any text following the identifier is ignored
Letters can be upper or lower case. Spaces and other non-letter
characters in the sequence are ignored. Letter U is translated to T.
All letters not equal to A, C, G, T or U are treated as unknown (N).
The sequences can be of any length.
All lines starting with `#' are treated as comment
lines, lines starting with `%%' may contain annotation (see below).
The execution time of the program is roughly proportional to the
Predict splice sites and start/stop codons
associated probabilities. The output
format is GFF
The signal prediction is different from most other predictors of
splice sites and start/stop, in that only signals that fit well into a
whole gene structure is predicted, i.e., the signals are not predicted
from the local sequence alone. This yields fewer predictions and
usually better, however, if there is an error that frameshifts an
actual gene or something like that, the splice sites might be missed
as well as the gene.
The predicted genes in a sequence are the most probable ones according
to the program (or rather the underlying hidden Markov model). It is
possible to also see suboptimal predictions. For instance, to see the
3 most probable predictions, hmmgene is run with `3 best predictions'
instead of `best prediction'.
The program will run approximately 3 times slower in that case.
Because of the slow-down of the program and the large amount of
information produced, it is best to use this option on a region, where
it is likely that there is only one gene. Then it will be possible to
see alternative ways of splicing it together. Although it is quite
possible that real alternative splicing can be predicted in this way,
this has not yet been investigated. Whether a gene is alternatively
spliced or not, it will often be usefull to see the alternative
possibilities that might score almost as well as the best prediction.
If something is known about one or more of the sequences, it can be
specified either in a separate annotation file or in the sequence
file. For instance if it is known that SEQ2 is non-coding from base
number 105 to 443, the annotation file must contain a line of the form
SEQ2 non-coding 105 443
Note that these keywords must appear exactly as written here (lower
An optional + or - at the end of a line indicates direct strand
(the direction of the sequence in the file) or the complementary
The same can be specified in the sequence file by preceeding each line
%% SEQ2 non-coding 105 443
This has to come before the actual sequence in the file, e.g.,
all annotation lines can come in the very beginning.
This is very useful if there are database hits to a sequence or
if repeats are mapped by some other program.
Assume for instance that there is a database hit to base
1503-1594 and alu repeats are found at position 10731-10890 and
13205-13356 in SEQ2. Then one might want to enter the lines
SEQ2 coding 1503 1594 +
SEQ2 non-coding 1503 1594 -
SEQ2 non-coding 10731 10890
SEQ2 non-coding 13205 13356
Here we indicated that the sequence is coding on the direct strand
from 1503 to 1594 and non-coding in this region on the complementary
strand. The two last lines means that the regions are non-coding
on BOTH STRANDS.
Regions specified in the file are not allowed to overlap except
on opposite strands.
If the annotation you give does not conform to the model, the program
will die. This happens for instance if the annotaion you give forces
- a non-consensus start or stop codon.
- a donor different from GT, or a an acceptor different from AG.
- start of an exon less than 25 bases from the beginning
of the sequence or an exon extending closer than 10 bases from
- a stop codon in a coding region.
For some reason the probability of the final exon is sometimes
larger than 1. Usually it is not very much. I can't find the
error. Please let me know if it happens in other cases.
If no start codon or stop codon is predicted for a gene (e.g.
begins and ends with an intron) the frame information and
scores might be wrong.
HMMgene can in principle predict a gene with a stop codon in frame,
if splicing happens in the middle of it. I have not yet seen
any examples though.
The output is a prediction of partial or complete genes in the
sequences.It is in
which is a sequence
annotation format developed with gene finding in mind. It is very
simple and therefore it is easy to develop programs in perl or awk to
post-process the output. The following is an
example of the form it takes with hmmgene.
Note that hmmgene only predicts coding regions. That is, the first
exon (`firstex' below) is only the coding part of the first coding
exon and similarly for the last exon (`lastex' below). Below a `gene'
therefore means the region of the gene from start to stop codon.
SEQ1 HMMgene1.1 firstex 692 702 0.347 + 2 bestparse:cds_1
SEQ1 HMMgene1.1 exon_1 2473 2711 0.421 + 1 bestparse:cds_1
SEQ1 HMMgene1.1 exon_2 2897 3081 0.544 + 0 bestparse:cds_1
SEQ1 HMMgene1.1 exon_3 10376 10563 0.861 + 2 bestparse:cds_1
SEQ1 HMMgene1.1 exon_4 11841 11891 0.857 + 2 bestparse:cds_1
SEQ1 HMMgene1.1 exon_5 12387 12483 0.993 + 0 bestparse:cds_1
SEQ1 HMMgene1.1 exon_6 13076 13211 0.970 + 1 bestparse:cds_1
SEQ1 HMMgene1.1 exon_7 13332 13415 0.926 + 1 bestparse:cds_1
SEQ1 HMMgene1.1 exon_8 13515 13603 1.000 + 0 bestparse:cds_1
SEQ1 HMMgene1.1 exon_9 14180 14235 1.000 + 2 bestparse:cds_1
SEQ1 HMMgene1.1 exon_10 14321 14408 0.999 + 0 bestparse:cds_1
SEQ1 HMMgene1.1 exon_11 14483 14579 0.877 + 1 bestparse:cds_1
SEQ1 HMMgene1.1 exon_12 14697 14764 0.639 + 0 bestparse:cds_1
SEQ1 HMMgene1.1 exon_13 14901 15030 0.835 + 1 bestparse:cds_1
SEQ1 HMMgene1.1 lastex 15643 15704 0.987 + 0 bestparse:cds_1
SEQ1 HMMgene1.1 CDS 692 15704 0.132 + . bestparse:cds_1
(the real list is tab separated)
- Sequence identifier
- Program name
- Prediction (see table below for the meaning).
- Score between 0 and 1
- Strand: $+$ for direct and $-$ for complementary
- Frame (for exons it is the position of the donor in the frame)
- Group to which prediction belong. If several CDS's are found
they will be called cds_1, cds_2, etc.
`bestparse:' is there because alternative predictions will
also be available (see below).
The score that comes with all the exons as well as the entire gene
`CDS' above) is a probability, so a value close to one means that
the program is fairly certain. (See `Known Bugs'.)
The program also outputs some comment lines which are preceeded by
|firstex || The coding part of the first coding exon starting with the first base of the start codon.
|exon_N || The N'th predicted internal coding exon.
|lastex || The coding part of the last coding exon ending with the last base of the stop codon.
|singleex || The coding part of an exon in a gene with only one coding exon.
|CDS || Coding region composed of the exon predictions prior to this line.
|START || Predicted start codon with position of first and last base (only with signal option).
|STOP || Predicted stop codon with position of first and last base (only with signals option).
|DON || Predicted donor site with position of the base before and after the splice site (only with signal option).
|ACC || Predicted acceptor site with position of the base before and after the splice site (only signal option).