Question: Has anyone run Phobius locally? Error - could not read provided fasta sequences
gravatar for s.kyungyong64
2.3 years ago by
Berkeley, USA
s.kyungyong6410 wrote:


I am currently trying to run Phobius ( in LInux to predict signal peptides in some proteins. I am not sure if anyone has run this recently, but please help me if you have!

After typing in the absolute path to the directory in

my $PHOBIUS_DIR = "/full/path/to/phobius";

In that directory, I typed in the command below. test.fasta existed in the same folder. Then, it returned

perl -short test.fasta

Phobius ver 1.01

(c) 2004 Lukas Kall, Anders Krogh, Erik Sonnhammer


Could not read provided fasta sequence at line 408.

Test.fasta looks like just usual a fasta file with >seq1 \n sequences

sequence • 1.5k views
ADD COMMENTlink modified 10 months ago by h.mon26k • written 2.3 years ago by s.kyungyong6410

I met the same problem, any solution so far?

ADD REPLYlink written 2.2 years ago by Guangyou Duan30

In my case, Duan's solution worked for me. I copied decodeanhmm.Linux_i686 from tmhmm-2.0c/bin to the Phobius folder as decodeanhmm.

ADD REPLYlink written 2.1 years ago by yongjoon0

You shouldn't need to alter the line:

my $PHOBIUS_DIR = "$FindBin::RealBin";

The FindBin module function is to locates the full path to the script bin directory, to allow the use of paths relative to the bin directory, so there should be no need to hard-code paths into the script.

ADD REPLYlink written 10 months ago by h.mon26k
gravatar for Guangyou Duan
2.2 years ago by
Guangyou Duan30 wrote:

It could be the problem of decodeanhmm, in my case I copy a new decodeanhmm from the installation directory of tmhmm-2.0c and it works.

ADD COMMENTlink written 2.2 years ago by Guangyou Duan30

This fixed it for me -


would result in an error:

 -bash: ./decodeanhmm: No such file or directory

(how is that even possible? the file is right there!), so I downloaded this copy:

and tried to run and got output immediately, instead of the above 'could not read provided fasta file'

In my case this was causing problems in the RGaugury pipeline, where the files with RLK and RLP candidate genes were empty because of phobius not working properly.

Edit: I now figured out what the No such file or directory error means. If you use the strings tool on the decodeanhmm binary that comes with phobius, you'll see this file pop up:


On my system that file doesn't exist, which causes the error. The decodeanhmm in the ParseTargs package doesn't have that string in its header, so it doesn't look for that file. The decodeanhmm.64bit file that comes with phobius also doesn't have that file, maybe renaming decodeanhmm.64bit to decodeanhmm (overwriting the 'original') fixes this problem too?

ADD REPLYlink modified 3 months ago • written 3 months ago by Philipp Bayer6.1k
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