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primers's Issues

Rename some of the parameters

opt_tm: float = 62.0
opt_gc: float = 0.5
opt_len: int = 22
penalty_tm: float = 1.0
penalty_gc: float = 3.0
penalty_len: float = 1.0
penalty_tm_diff: float = 1.0
penalty_dg: float = 2.0
penalty_offtarget: float = 20.0

to

optimal_tm: float = 62.0
optimal_gc: float = 0.5
optimal_len: int = 22
penalty_tm_delta: float = 1.0 # delta from optimal
penalty_gc_delta: float = 3.0
penalty_len_delta: float = 1.0
penalty_dg_delta: float = 2.0
penalty_offtarget: float = 20.0
penalty_pair_tm_delta: float = 1.0 # delta of tm between the pair's primers

released package on pypi don't contain `requirements.txt` which makes installation fail if not using `whl` file

to reproduce

wget https://pypi.io/packages/source/p/primers/primers-0.5.4.tar.gz
tar zxvf primer*
cd primers-0.5.4
ls

got me

LICENSE
PKG-INFO
primers
primers.egg-info
README.md
setup.cfg
setup.py
tests

you see there is no requirements.txt, but setup.py reads it.

thus got me

......
  Installing build dependencies ... done
  Running command Getting requirements to build wheel
  <string>:3: DeprecationWarning: pkg_resources is deprecated as an API. See https://setuptools.pypa.io/en/latest/pkg_resources.html
  Traceback (most recent call last):
    File "/tmp/primers-0.5.4/.venv/lib/python3.11/site-packages/pip/_vendor/pyproject_hooks/_in_process/_in_process.py", line 353, in <module>
      main()
    File "/tmp/primers-0.5.4/.venv/lib/python3.11/site-packages/pip/_vendor/pyproject_hooks/_in_process/_in_process.py", line 335, in main
      json_out['return_val'] = hook(**hook_input['kwargs'])
                               ^^^^^^^^^^^^^^^^^^^^^^^^^^^^
    File "/tmp/primers-0.5.4/.venv/lib/python3.11/site-packages/pip/_vendor/pyproject_hooks/_in_process/_in_process.py", line 118, in get_requires_for_build_wheel
      return hook(config_settings)
             ^^^^^^^^^^^^^^^^^^^^^
    File "/tmp/pip-build-env-hoe006rw/overlay/lib/python3.11/site-packages/setuptools/build_meta.py", line 325, in get_requires_for_build_wheel
      return self._get_build_requires(config_settings, requirements=['wheel'])
             ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
    File "/tmp/pip-build-env-hoe006rw/overlay/lib/python3.11/site-packages/setuptools/build_meta.py", line 295, in _get_build_requires
      self.run_setup()
    File "/tmp/pip-build-env-hoe006rw/overlay/lib/python3.11/site-packages/setuptools/build_meta.py", line 487, in run_setup
      super().run_setup(setup_script=setup_script)
    File "/tmp/pip-build-env-hoe006rw/overlay/lib/python3.11/site-packages/setuptools/build_meta.py", line 311, in run_setup
      exec(code, locals())
    File "<string>", line 10, in <module>
  FileNotFoundError: [Errno 2] No such file or directory: 'requirements.txt'
  error: subprocess-exited-with-error
......

the reason why I don't use wheel is that I am packing it to bioconda conda channel. patching it as temp workaround

.seq property of reverse primer is str type while forward primer is Bio.Seq type

This is not a dealbreaker but having the reverse complement method

def _rc(seq: str) -> str:
return str means that the underlying 'seq' property for the reverse primer is also a string. This differs from the seq property of the forward primer, which is a Bio.Seq type.
I think the seq properties for both primers should be either a str of Bio.Seq type.
Not the end of the world but has made it a little frustrating dealing with this property.

Error when running off-target example

I am getting the following error when trying to run the off-target example:

% primers AATGAGACAATAGCACACACAGCTAGGTCAGCATACGAAA -t ggaattacgtAATGAGACAATAGCACACACAGCTAGGTCAGCATACGAAAggaccagttacagga
Traceback (most recent call last):
File "/opt/anaconda3/bin/primers", line 8, in
sys.exit(run())
File "/opt/anaconda3/lib/python3.9/site-packages/primers/main.py", line 19, in run
fwd, rev = primers(
File "/opt/anaconda3/lib/python3.9/site-packages/primers/primers.py", line 258, in primers
fwd_primers = _primers(
File "/opt/anaconda3/lib/python3.9/site-packages/primers/primers.py", line 314, in _primers
ot = offtargets(seq, offtarget_check)
File "/opt/anaconda3/lib/python3.9/site-packages/primers/offtargets.py", line 34, in offtargets
for m in mutate(check_seq[s : s + 10]):
File "/opt/anaconda3/lib/python3.9/site-packages/primers/offtargets.py", line 29, in mutate
for m in mutate_map[c]:
KeyError: 'g'

Standalone tool to evaluate PCR quality

It would be great to be able to score a PCR based on the primers that I specify. Something like this:

primers -t {template} -oligo1 {oligo1_seq} -oligo2 {oligo2_seq}

Thanks,
Joe

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