pax_global_header00006660000000000000000000000064141250247010014506gustar00rootroot0000000000000052 comment=ddd93aaf26e34986b23c19988392f190672b86a3 InSilicoSeq-1.5.4/000077500000000000000000000000001412502470100136775ustar00rootroot00000000000000InSilicoSeq-1.5.4/.codecov.yml000066400000000000000000000001451412502470100161220ustar00rootroot00000000000000ignore: - "**/test/*.py" # ignore test scripts - "**/app.py" # ignore main application script InSilicoSeq-1.5.4/.github/000077500000000000000000000000001412502470100152375ustar00rootroot00000000000000InSilicoSeq-1.5.4/.github/workflows/000077500000000000000000000000001412502470100172745ustar00rootroot00000000000000InSilicoSeq-1.5.4/.github/workflows/pythonpackage.yml000066400000000000000000000020311412502470100226500ustar00rootroot00000000000000name: CI on: [push] jobs: build: runs-on: ubuntu-latest strategy: max-parallel: 4 matrix: python-version: [3.6, 3.7, 3.8] steps: - uses: actions/checkout@v1 - name: Set up Python ${{ matrix.python-version }} uses: actions/setup-python@v1 with: python-version: ${{ matrix.python-version }} - name: Install dependencies run: | python -m pip install --upgrade pip pip install pipenv pipenv install --dev - name: Test with nose run: | chmod -w data/read_only.fasta pipenv run tests - name: Upload to Codecov uses: codecov/codecov-action@v1.0.2 with: token: ${{ secrets.CODECOV_TOKEN }} file: coverage.xml - name: Test install run: | pip install -e . InSilicoSeq-1.5.4/.gitignore000066400000000000000000000010161412502470100156650ustar00rootroot00000000000000# project specific files *.fastq *.bam *.bam.bai *.bt2 test_runs/ # Byte-compiled / optimized / DLL files __pycache__/ *.pyc # Distribution / packaging *.egg-info/ dist/ build/ # Unit test / coverage reports .cache .coverage coverage.xml # files needed for tests !SRR1660402_mapped.bam !SRR1660402_mapped.bam.bai !ecoli.fasta !ecoli_R*.fastq !ecoli.bam !ecoli.bam.bai !substitutions_test.bam !substitutions_test.bam.bai # virtualenv venv_iss/ # MacOS stuff .DS_Store # fastqc reports *fastqc* # editor configs .vscode/InSilicoSeq-1.5.4/.readthedocs.yml000066400000000000000000000001301412502470100167570ustar00rootroot00000000000000python: install: - requirements: doc/requirements.txt - method: pip InSilicoSeq-1.5.4/CODE_OF_CONDUCT.md000066400000000000000000000062361412502470100165050ustar00rootroot00000000000000# Contributor Covenant Code of Conduct ## Our Pledge In the interest of fostering an open and welcoming environment, we as contributors and maintainers pledge to making participation in our project and our community a harassment-free experience for everyone, regardless of age, body size, disability, ethnicity, gender identity and expression, level of experience, nationality, personal appearance, race, religion, or sexual identity and orientation. ## Our Standards Examples of behavior that contributes to creating a positive environment include: * Using welcoming and inclusive language * Being respectful of differing viewpoints and experiences * Gracefully accepting constructive criticism * Focusing on what is best for the community * Showing empathy towards other community members Examples of unacceptable behavior by participants include: * The use of sexualized language or imagery and unwelcome sexual attention or advances * Trolling, insulting/derogatory comments, and personal or political attacks * Public or private harassment * Publishing others' private information, such as a physical or electronic address, without explicit permission * Other conduct which could reasonably be considered inappropriate in a professional setting ## Our Responsibilities Project maintainers are responsible for clarifying the standards of acceptable behavior and are expected to take appropriate and fair corrective action in response to any instances of unacceptable behavior. Project maintainers have the right and responsibility to remove, edit, or reject comments, commits, code, wiki edits, issues, and other contributions that are not aligned to this Code of Conduct, or to ban temporarily or permanently any contributor for other behaviors that they deem inappropriate, threatening, offensive, or harmful. ## Scope This Code of Conduct applies both within project spaces and in public spaces when an individual is representing the project or its community. Examples of representing a project or community include using an official project e-mail address, posting via an official social media account, or acting as an appointed representative at an online or offline event. Representation of a project may be further defined and clarified by project maintainers. ## Enforcement Instances of abusive, harassing, or otherwise unacceptable behavior may be reported by contacting the project team at [INSERT EMAIL ADDRESS]. All complaints will be reviewed and investigated and will result in a response that is deemed necessary and appropriate to the circumstances. The project team is obligated to maintain confidentiality with regard to the reporter of an incident. Further details of specific enforcement policies may be posted separately. Project maintainers who do not follow or enforce the Code of Conduct in good faith may face temporary or permanent repercussions as determined by other members of the project's leadership. ## Attribution This Code of Conduct is adapted from the [Contributor Covenant][homepage], version 1.4, available at [http://contributor-covenant.org/version/1/4][version] [homepage]: http://contributor-covenant.org [version]: http://contributor-covenant.org/version/1/4/ InSilicoSeq-1.5.4/CONTRIBUTING.md000066400000000000000000000017251412502470100161350ustar00rootroot00000000000000# Contributing to InSilicoSeq Thank you for your interest in our software :heart: :tada: ## Conduct We are committed to providing a friendly, safe and welcoming environment for all. If you wish to contribute, you are expected to adhere to a [code of conduct](CODE_OF_CONDUCT.md) ## How to contribute This project is still at an early stage and open to suggestions of any kind. Chat with us by [opening an issue](https://github.com/HadrienG/InSilicoSeq/issues) or fork the repository and [open a pull request](https://github.com/HadrienG/InSilicoSeq/pulls) ## Coding guidelines * Please adhere to [PEP8](https://www.python.org/dev/peps/pep-0008/) * The docstrings should follow the [Google style guide](http://www.sphinx-doc.org/en/stable/ext/napoleon.html) * Your pull request should pass the python3 tests, and be backwards compatible with python2. New code/functionality should come with a test. * Ideally a pull request should solve one problem or add one functionality. InSilicoSeq-1.5.4/Dockerfile000066400000000000000000000002341412502470100156700ustar00rootroot00000000000000FROM python:3 LABEL maintainer "Hadrien Gourlé " WORKDIR /usr/src/app COPY . . RUN pip install --no-cache-dir . CMD [ "iss" ] InSilicoSeq-1.5.4/LICENSE000066400000000000000000000020601412502470100147020ustar00rootroot00000000000000MIT License Copyright (c) 2016 Hadrien Gourlé Permission is hereby granted, free of charge, to any person obtaining a copy of this software and associated documentation files (the "Software"), to deal in the Software without restriction, including without limitation the rights to use, copy, modify, merge, publish, distribute, sublicense, and/or sell copies of the Software, and to permit persons to whom the Software is furnished to do so, subject to the following conditions: The above copyright notice and this permission notice shall be included in all copies or substantial portions of the Software. THE SOFTWARE IS PROVIDED "AS IS", WITHOUT WARRANTY OF ANY KIND, EXPRESS OR IMPLIED, INCLUDING BUT NOT LIMITED TO THE WARRANTIES OF MERCHANTABILITY, FITNESS FOR A PARTICULAR PURPOSE AND NONINFRINGEMENT. IN NO EVENT SHALL THE AUTHORS OR COPYRIGHT HOLDERS BE LIABLE FOR ANY CLAIM, DAMAGES OR OTHER LIABILITY, WHETHER IN AN ACTION OF CONTRACT, TORT OR OTHERWISE, ARISING FROM, OUT OF OR IN CONNECTION WITH THE SOFTWARE OR THE USE OR OTHER DEALINGS IN THE SOFTWARE. InSilicoSeq-1.5.4/MANIFEST.in000066400000000000000000000000511412502470100154310ustar00rootroot00000000000000include README.md include iss/profiles/* InSilicoSeq-1.5.4/Pipfile000066400000000000000000000006111412502470100152100ustar00rootroot00000000000000[[source]] url = "https://pypi.python.org/simple" verify_ssl = true name = "pypi" [packages] future = "*" numpy = "*" scipy = "*" biopython = "==1.78" joblib = "*" pysam = "==0.15.4" requests = "*" urllib3 = ">=1.26.5" [dev-packages] nose = "*" codecov = "*" "pep8" = "*" pycodestyle = "*" [scripts] iss = "python -m iss" tests = "nosetests --with-coverage --cover-package=iss --cover-xml" InSilicoSeq-1.5.4/Pipfile.lock000066400000000000000000000457561412502470100161620ustar00rootroot00000000000000{ "_meta": { "hash": { "sha256": "9325d4461986d06c73ae5a49a7cef5fff6276280f76e9c9141433fc884c0f395" }, "pipfile-spec": 6, "requires": {}, "sources": [ { "name": "pypi", "url": "https://pypi.python.org/simple", "verify_ssl": true } ] }, 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[![doi](https://img.shields.io/badge/doi-10.1093%2Fbioinformatics%2Fbty630-blue.svg)](https://doi.org/10.1093/bioinformatics/bty630) [![LICENSE](https://img.shields.io/badge/license-MIT-lightgrey.svg)](LICENSE) InSilicoSeq is a sequencing simulator producing realistic Illumina reads. Primarily intended for simulating metagenomic samples, it can also be used to produce sequencing data from a single genome. InSilicoSeq is written in python, and use kernel density estimators to model the read quality of real sequencing data. InSilicoSeq support substitution, insertion and deletion errors. If you don't have the use for insertion and deletion error a basic error model is provided. ## Installation Insilicoseq is Available in [bioconda](https://bioconda.github.io/). To install with conda: ```shell conda install -c bioconda insilicoseq ``` Or with pip: ```shell pip install InSilicoSeq ``` _Note:_ Insilicoseq requires python >= 3.5 Alternatively, with docker: ```shell docker pull hadrieng/insilicoseq:latest ``` For more installation options, please refer to the full [documentation](http://insilicoseq.readthedocs.io) ## Usage InSilicoSeq has two subcommands: `iss generate` to generate Illumina reads and `iss model` to create an error model from which the reads will take their characteristics. InSilicoSeq comes with pre-computed error models that should be sufficient for most use cases. ### Generate reads with a pre-computed error model for generating 1 million reads modelling a MiSeq instrument: ```shell curl -O -J -L https://osf.io/thser/download # download the example data iss generate --genomes SRS121011.fasta --model miseq --output miseq_reads ``` where `genomes.fasta` should be replaced by a (multi-)fasta file containing the reference genome(s) from which the simulated reads will be generated. InSilicoSeq comes with 3 error models: `MiSeq`, `HiSeq` and `NovaSeq`. If you have built your own model, pass the `.npz` file to the `--model` argument to simulate reads from your own error model. For 10 million reads and a custom error model: ```shell curl -O -J -L https://osf.io/thser/download # download the example data iss generate -g SRS121011.fasta -n 10m --model my_model.npz --output /path/to/my_reads ``` granted you have built `my_model.npz` with `iss model` (see [below](#create-your-own-error-model)) For more examples and a full list of options, please refer to the full [documentation](http://insilicoseq.readthedocs.io) ### Generate reads without input genomes We can download some for you! InSilicoSeq can download random genomes from the ncbi using the infamous [eutils](https://www.ncbi.nlm.nih.gov/books/NBK25501/) The command ```shell iss generate --ncbi bacteria -u 10 --model MiSeq --output ncbi_reads ``` will generate 1 million reads from 10 random bacterial genomes. For more examples and a full list of options, please refer to the full [documentation](http://insilicoseq.readthedocs.io) ### Create your own error model If you do not wish to use the pre-computed error models provided with InSilicoSeq, it is possible to create your own. Say you have a reference metagenome called `genomes.fasta`, and read pairs `reads_R1.fastq.gz` and `reads_R2.fastq.gz` Align you reads against the reference: ```shell bowtie2-build genomes.fasta genomes bowtie2 -x genomes -1 reads_R1.fastq.gz -2 reads_R2.fastq.gz | \ samtools view -bS | samtools sort -o genomes.bam samtools index genomes.bam ``` then build the model: ```shell iss model -b genomes.bam -o genomes ``` which will create a `genome.npz` file containing your newly built model ## License Code is under the [MIT](LICENSE) license. ## Issues Found a bug or have a question? Please open an [issue](https://github.com/HadrienG/InSilicoSeq/issues) ## Contributing We welcome contributions from the community! See our [Contributing](CONTRIBUTING.md) guidelines ## Citation If you use our software, please cite us! > Gourlé H, Karlsson-Lindsjö O, Hayer J and Bongcam+Rudloff E, Simulating Illumina data with InSilicoSeq. _Bioinformatics_ (2018) doi:10.1093/bioinformatics/bty630 InSilicoSeq-1.5.4/data/000077500000000000000000000000001412502470100146105ustar00rootroot00000000000000InSilicoSeq-1.5.4/data/abundance.txt000066400000000000000000000001201412502470100172620ustar00rootroot00000000000000genome_A 0.2 genome_T 0.2 genome_GC 0.4 genome_ATCG 0.1 genome_TA 0.1 InSilicoSeq-1.5.4/data/bad_abundance.txt000066400000000000000000000001151412502470100200740ustar00rootroot00000000000000genome_A 0.2 genome_T 0.2 genome_GC0.4 genome_ATCG 0.1 genome_TA 0.1 InSilicoSeq-1.5.4/data/draft.fasta000066400000000000000000000002151412502470100167260ustar00rootroot00000000000000>contig_1 length=29 AAAAAAAAAAAAAAAAAAAAAAAAAAAAA >contig_2 length=12 AAATTTCCCCCC >contig_3 length=37 CCCCCCCCCCAAAAAAAAAATTTTTTTTTTGGGGGGG InSilicoSeq-1.5.4/data/ecoli.bam000066400000000000000000000013201412502470100163600ustar00rootroot00000000000000BCM 0EEY̪ZA[Ik@h45"aNƫ̧pINײtM%cb8c%\Len_ p.ejO+(!4*O[29WF߽\GMKg ;NNn/ҏ:]/_ HBCN@rn6fO&m8l<D8&H"\Ԡ/`Ջz>>!!8G, l2 ;>jcJ5!L=rn/sh0 qQtd#gu1ԑGvv ZmeV6XEŊr>dVYzdI){O6m8o|QCzRȝb5j2}4pQ4 ]Ȗ2e1?Ƭ9V/x5V9˛ ӟ]4~ư#QASӤɃ/\swÚerneh${X}k2u1r8y9֘9Uޣ{`xlNC_002695.1 Escherichia coli O157:H7 str. Sakai chromosome, 1330 first bases AGCTTTTCATTCTGACTGCAACGGGCAATATGTCTCTGTGTGGATTAAAAAAAGAGTCTCTGACAGCAGC TTCTGAACTGGTTACCTGCCGTGAGTAAATTAAAATTTTATTGACTTAGGTCACTAAATACTTTAACCAA TATAGGCATAGCGCACAGACAGATAAAAATTACAGAGTACACAACATCCATGAAACGCATTAGCACCACC ATTACCACCACCATCACCACCACCATCACCATTACCATTACCACAGGTAACGGTGCGGGCTGACGCGTAC AGGAAACACAGAAAAAAGCCCGCACCTGACAGTGCGGGCTTTTTTTTCGACCAAAGGTAACGAGGTAACA ACCATGCGAGTGTTGAAGTTCGGCGGTACATCAGTGGCAAATGCAGAACGTTTTCTGCGGGTTGCCGATA TTCTGGAAAGCAATGCCAGGCAGGGGCAGGTGGCCACCGTCCTCTCTGCCCCCGCCAAAATCACCAACCA CCTGGTGGCGATGATTGAAAAAACCATTAGCGGCCAGGATGCTTTACCCAATATCAGCGATGCCGAACGT ATTTTTGCCGAACTTCTGACGGGACTCGCCGCCGCCCAGCCGGGATTCCCGCTGGCGCAATTGAAAACTT TCGTCGACCAGGAATTTGCCCAAATAAAACATGTCCTGCATGGCATTAGTTTGTTAGGGCAGTGCCCGGA TAGCATTAACGCTGCGCTGATTTGCCGTGGCGAGAAAATGTCGATCGCCATTATGGCCGGCGTATTAGAA GCGCGCGGTCACAACGTTACCGTTATCGATCCGGTCGAAAAACTGCTGGCAGTGGGGCATTACCTCGAAT CTACTGTCGATATTGCAGAGTCCACCCGCCGTATTGCGGCAAGTCGTATTCCGGCTGATCACATGGTGCT GATGGCAGGTTTCACCGCCGGTAATGAAAAAGGCGAACTGGTGGTACTTGGACGCAACGGTTCCGACTAC TCCGCGGCGGTGCTGGCTGCCTGTTTACGCGCCGATTGTTGCGAGATTTGGACGGACGTTGACGGGGTAT ATACCTGCGACCCGCGTCAGGTGCCCGATGCGAGGTTGTTGAAATCGATGTCCTACCAGGAAGCGATGGA GCTTTCCTACTTCGGCGCTAAAGTTCTTCACCCCCGCACCATTACCCCCATCGCCCAGTTCCAGATCCCT TGCCTGATTAAAAATACCGGAAATCCTCAAGCTCCAGGTACGCTCATTGGTGCCAGTCGTGATGAAGACG AATTACCGGTCAAGGGCATTTCCAATCTGAATAATATGGCAATGTTCAGCGTTTCCGGCCCGGGGATGAA InSilicoSeq-1.5.4/data/ecoli.npz000066400000000000000000000466301412502470100164450ustar00rootroot00000000000000PK0lfK2 LK\ model.npyPZ\nnj_TR_ wK)Ng$: (+!SPK0lfK7qEXread_length.npyPZ\nni_TR_ wK)Ng$: (KPK0lfKcinsert_size.npy5Ww8^ )De$ EEJ[А2"~e-2BFLB[^~_^{u]wy~ι<\gD\5^v[EZi1i6.n.淯;XY/zw5w^{<(Wvok<<[%΢sU?jDl"-Hr^v+\q(̌VW$i/TJ#\ 1sjx1>s_< 7! 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Gb JDJ!q 3Zc qq"ӟ q ӟ*4BΌ@ę8F98C/bB\Œ.Ee@ĕF5鯓BȌ#&B VB#BwI q!eF!~B<"!B<!e$qxO24!ag!g"!^b%HWkxo0xo3;x1B|HLj2g_0_37B|G?1ψ2o0BMѿش!4'#IikD:?CdlJeY! /Ay|=:a.p~8±tYsam滬 F9Bu4?X\ֱpc::e`catZP/R3RbϮvmbFDrΏrrٖ/.+73,O3y 9S@ZnG!")47dpXrHE3E RE(M(rG T?˱ɑ6rn; \bbb␻7w*.ʮTq1U ;^Or"~Gz |(c}O$)3 _ž4J_O~Я~'/aɊV1/il*|IvKq.-Ot2q2Wrr*2Wʜ"rNu5»jwRnm[s=i\4; 972j&5e.r3ׂcûZw)5mm˹qW{t`#;8w6WN溲Ӎsw\/vzs׼r3?)g0sCqsj5L9%3.RHF3]xv'ph5I9Kag gqWr̥n5U9~2ؙyzxWyWrg]r̅<økrf1lsS\vq޵kr1B¸H9K[2ˍV(g%sYyqZcn=;8o d޵Y[6ۍv(g'syq^cn?;84:a(c]ͻN($=qYc<;8_4]a*k]וs|[;35/PK0lfK8tins_reverse.npyuexKmR{@ ViJ mڳIBwuwwwwܝ/s+=ɼo_Y8zqǜ4_ktOI϶B'm ̠>ϮvmbFDrΏrrٖ/.+73,O3y 9S@ZnG!")47dpXrHE3E RE(M(rG T?˱ɑ6rn; \bbb␻7w*.ʮTq1U ;^Or"~Gz |(c}O$)3 _ž4J_O~Я~'/aɊV1/il*|IvKq.-Ot2q2Wrr*2Wʜ"rNu5»jwRnm[s=i\4; 972j&5e.r3ׂcûZw)5mm˹qW{t`#;8w6WN溲Ӎsw\/vzs׼r3?)g0sCqsj5L9%3.RHF3]xv'ph5I9Kag gqWr̥n5U9~2ؙyzxWyWrg]r̅<økrf1lsS\vq޵kr1B¸H9K[2ˍV(g%sYyqZcn=;8o d޵Y[6ۍv(g'syq^cn?;84:a(c]ͻN($=qYc<;8_4]a*k]וs|[;35/PK0lfK8tdel_forward.npyuexKmR{@ ViJ 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*subst_choices_forward.npyPK0lfKG@:.subst_choices_reverse.npyPK0lfK8t3ins_forward.npyPK0lfK8tX9ins_reverse.npyPK0lfK8t>del_forward.npyPK0lfK8tDdel_reverse.npyPK G;JInSilicoSeq-1.5.4/data/ecoli_R1.fastq000066400000000000000000000013171412502470100173070ustar00rootroot00000000000000@read_1_1 perfect i_size=10 AGCTTCTGAACTGGTTACCT + IIIIIIIIIIIIIIIIIIII @read_2_1 subst_C_19 i_size=10 ATAGGCATAGCGCACAGAtA + IIIIIIIIIIIIIIIIIIII @read_3_1 low_q i_size=10 CTGCCGTGAGTAAATTAAAA + ???????????????????? @read_4_1 ins_9 i_size=10 GATCCCTTtGCCTGATTAAA + IIIIIIIIIIIIIIIIIIII @read_5_1 perfect i_size=10 TACCCAATATCAGCGATGCC + IIIIIIIIIIIIIIIIIIII @read_6_1 perfect i_size=10 AGCTTCTGAACTGGTTACCT + IIIIIIIIIIIIIIIIIIII @read_7_1 subst_C_19 i_size=10 ATAGGCATAGCGCACAGAtA + IIIIIIIIIIIIIIIIIIII @read_8_1 low_q i_size=10 CTGCCGTGAGTAAATTAAAA + ???????????????????? @read_9_1 ins_9 i_size=10 GATCCCTTtGCCTGATTAAA + IIIIIIIIIIIIIIIIIIII @read_10_1 perfect i_size=10 TACCCAATATCAGCGATGCC + IIIIIIIIIIIIIIIIIIII InSilicoSeq-1.5.4/data/ecoli_R2.fastq000066400000000000000000000013311412502470100173040ustar00rootroot00000000000000@read_1_2 subst_T_20 i_size=10 AAGTCAATAAAATTTTAATa + IIIIIIIIIIIIIIIIII++ @read_2_2 perfect i_size=10 ATGGATGTTGTGTACTCTGT + IIIIIIIIIIIIIIIIII++ @read_3_2 low_q i_size=10 TAAAGTATTTAGTGACCTAA + ??????????????????55 @read_4_2 perfect i_size=10 GCGTACCTGGAGCTTGAGGA + IIIIIIIIIIIIIIIIII++ @read_5_2 deletion_4 i_size=10 TCCGTCAGAAGTTCGGCAAA + IIIIIIIIIIIIIIIIII++ @read_6_2 subst_T_20 i_size=10 AAGTCAATAAAATTTTAATa + IIIIIIIIIIIIIIIIII++ @read_7_2 perfect i_size=10 ATGGATGTTGTGTACTCTGT + IIIIIIIIIIIIIIIIII++ @read_8_2 low_q i_size=10 TAAAGTATTTAGTGACCTAA + ??????????????????55 @read_9_2 perfect i_size=10 GCGTACCTGGAGCTTGAGGA + IIIIIIIIIIIIIIIIII++ @read_10_2 deletion_4 i_size=10 TCCGTCAGAAGTTCGGCAAA + IIIIIIIIIIIIIIIIII++ InSilicoSeq-1.5.4/data/empty_file000066400000000000000000000000001412502470100166560ustar00rootroot00000000000000InSilicoSeq-1.5.4/data/genomes.fasta000066400000000000000000000067701412502470100172770ustar00rootroot00000000000000>genome_A length=1460 AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA AAAAAAAAAAAAAAAAAAAA >genome_T length=640 TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT TTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTTT >genome_GC length=600 GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC GCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGCGC >genome_ATCG length=425 ATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCG ATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCG ATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCG ATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCG ATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCGATCG ATCGATCGATCGATCGATCG >genome_TA length=300 TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA TATATATATATATATATATATATATATATATATATATATATATATATATATATATATATA InSilicoSeq-1.5.4/data/read_only.fasta000066400000000000000000000054461412502470100176150ustar00rootroot00000000000000>NC_002695.1 Escherichia coli O157:H7 str. Sakai chromosome, 1330 first bases AGCTTTTCATTCTGACTGCAACGGGCAATATGTCTCTGTGTGGATTAAAAAAAGAGTCTCTGACAGCAGC TTCTGAACTGGTTACCTGCCGTGAGTAAATTAAAATTTTATTGACTTAGGTCACTAAATACTTTAACCAA TATAGGCATAGCGCACAGACAGATAAAAATTACAGAGTACACAACATCCATGAAACGCATTAGCACCACC ATTACCACCACCATCACCACCACCATCACCATTACCATTACCACAGGTAACGGTGCGGGCTGACGCGTAC AGGAAACACAGAAAAAAGCCCGCACCTGACAGTGCGGGCTTTTTTTTCGACCAAAGGTAACGAGGTAACA ACCATGCGAGTGTTGAAGTTCGGCGGTACATCAGTGGCAAATGCAGAACGTTTTCTGCGGGTTGCCGATA TTCTGGAAAGCAATGCCAGGCAGGGGCAGGTGGCCACCGTCCTCTCTGCCCCCGCCAAAATCACCAACCA CCTGGTGGCGATGATTGAAAAAACCATTAGCGGCCAGGATGCTTTACCCAATATCAGCGATGCCGAACGT ATTTTTGCCGAACTTCTGACGGGACTCGCCGCCGCCCAGCCGGGATTCCCGCTGGCGCAATTGAAAACTT TCGTCGACCAGGAATTTGCCCAAATAAAACATGTCCTGCATGGCATTAGTTTGTTAGGGCAGTGCCCGGA TAGCATTAACGCTGCGCTGATTTGCCGTGGCGAGAAAATGTCGATCGCCATTATGGCCGGCGTATTAGAA GCGCGCGGTCACAACGTTACCGTTATCGATCCGGTCGAAAAACTGCTGGCAGTGGGGCATTACCTCGAAT CTACTGTCGATATTGCAGAGTCCACCCGCCGTATTGCGGCAAGTCGTATTCCGGCTGATCACATGGTGCT GATGGCAGGTTTCACCGCCGGTAATGAAAAAGGCGAACTGGTGGTACTTGGACGCAACGGTTCCGACTAC TCCGCGGCGGTGCTGGCTGCCTGTTTACGCGCCGATTGTTGCGAGATTTGGACGGACGTTGACGGGGTAT ATACCTGCGACCCGCGTCAGGTGCCCGATGCGAGGTTGTTGAAATCGATGTCCTACCAGGAAGCGATGGA GCTTTCCTACTTCGGCGCTAAAGTTCTTCACCCCCGCACCATTACCCCCATCGCCCAGTTCCAGATCCCT TGCCTGATTAAAAATACCGGAAATCCTCAAGCTCCAGGTACGCTCATTGGTGCCAGTCGTGATGAAGACG AATTACCGGTCAAGGGCATTTCCAATCTGAATAATATGGCAATGTTCAGCGTTTCCGGCCCGGGGATGAA >NC_002695.1 Escherichia coli O157:H7 str. Sakai chromosome, 1330 first bases AGCTTTTCATTCTGACTGCAACGGGCAATATGTCTCTGTGTGGATTAAAAAAAGAGTCTCTGACAGCAGC TTCTGAACTGGTTACCTGCCGTGAGTAAATTAAAATTTTATTGACTTAGGTCACTAAATACTTTAACCAA TATAGGCATAGCGCACAGACAGATAAAAATTACAGAGTACACAACATCCATGAAACGCATTAGCACCACC ATTACCACCACCATCACCACCACCATCACCATTACCATTACCACAGGTAACGGTGCGGGCTGACGCGTAC AGGAAACACAGAAAAAAGCCCGCACCTGACAGTGCGGGCTTTTTTTTCGACCAAAGGTAACGAGGTAACA ACCATGCGAGTGTTGAAGTTCGGCGGTACATCAGTGGCAAATGCAGAACGTTTTCTGCGGGTTGCCGATA TTCTGGAAAGCAATGCCAGGCAGGGGCAGGTGGCCACCGTCCTCTCTGCCCCCGCCAAAATCACCAACCA CCTGGTGGCGATGATTGAAAAAACCATTAGCGGCCAGGATGCTTTACCCAATATCAGCGATGCCGAACGT ATTTTTGCCGAACTTCTGACGGGACTCGCCGCCGCCCAGCCGGGATTCCCGCTGGCGCAATTGAAAACTT TCGTCGACCAGGAATTTGCCCAAATAAAACATGTCCTGCATGGCATTAGTTTGTTAGGGCAGTGCCCGGA TAGCATTAACGCTGCGCTGATTTGCCGTGGCGAGAAAATGTCGATCGCCATTATGGCCGGCGTATTAGAA GCGCGCGGTCACAACGTTACCGTTATCGATCCGGTCGAAAAACTGCTGGCAGTGGGGCATTACCTCGAAT CTACTGTCGATATTGCAGAGTCCACCCGCCGTATTGCGGCAAGTCGTATTCCGGCTGATCACATGGTGCT GATGGCAGGTTTCACCGCCGGTAATGAAAAAGGCGAACTGGTGGTACTTGGACGCAACGGTTCCGACTAC TCCGCGGCGGTGCTGGCTGCCTGTTTACGCGCCGATTGTTGCGAGATTTGGACGGACGTTGACGGGGTAT ATACCTGCGACCCGCGTCAGGTGCCCGATGCGAGGTTGTTGAAATCGATGTCCTACCAGGAAGCGATGGA GCTTTCCTACTTCGGCGCTAAAGTTCTTCACCCCCGCACCATTACCCCCATCGCCCAGTTCCAGATCCCT TGCCTGATTAAAAATACCGGAAATCCTCAAGCTCCAGGTACGCTCATTGGTGCCAGTCGTGATGAAGACG AATTACCGGTCAAGGGCATTTCCAATCTGAATAATATGGCAATGTTCAGCGTTTCCGGCCCGGGGATGAA InSilicoSeq-1.5.4/data/substitutions_test.bam000066400000000000000000000012411412502470100212650ustar00rootroot00000000000000BCMj0)>qU խ୤]6Y%}=:ŁB%*e[U{>l;d- U!bRL$Iw}?I&قk5v g9Wz~5tۚFxS{g[x,ĢLpxO٧#r dwcB.V#BCj@/KrIKuP]}N"8>qD9ӳ^[~_C.;@"nq ODZ=ѵs} ] |w#`=(XWbWId&y9>ʰ/Y}˱>6u~8)uZb k* v%8!mMR,]\]#LGI1 Vok|9w:1E>V}_o]Fy"zͫ5'([8" U-W y/v()~LDL-!YR6`#@BUtrFE;eWBCInSilicoSeq-1.5.4/data/substitutions_test.bam.bai000066400000000000000000000001401412502470100220140ustar00rootroot00000000000000BAIIJ InSilicoSeq-1.5.4/doc/000077500000000000000000000000001412502470100144445ustar00rootroot00000000000000InSilicoSeq-1.5.4/doc/Makefile000066400000000000000000000011431412502470100161030ustar00rootroot00000000000000# Minimal makefile for Sphinx documentation # # You can set these variables from the command line. SPHINXOPTS = SPHINXBUILD = python -msphinx SPHINXPROJ = InSilicoSeq SOURCEDIR = . BUILDDIR = _build # Put it first so that "make" without argument is like "make help". help: @$(SPHINXBUILD) -M help "$(SOURCEDIR)" "$(BUILDDIR)" $(SPHINXOPTS) $(O) .PHONY: help Makefile # Catch-all target: route all unknown targets to Sphinx using the new # "make mode" option. $(O) is meant as a shortcut for $(SPHINXOPTS). %: Makefile @$(SPHINXBUILD) -M $@ "$(SOURCEDIR)" "$(BUILDDIR)" $(SPHINXOPTS) $(O)InSilicoSeq-1.5.4/doc/conf.py000066400000000000000000000114331412502470100157450ustar00rootroot00000000000000#!/usr/bin/env python3 # -*- coding: utf-8 -*- # # InSilicoSeq documentation build configuration file, created by # sphinx-quickstart on Tue May 30 11:45:01 2017. # # This file is execfile()d with the current directory set to its # containing dir. # # Note that not all possible configuration values are present in this # autogenerated file. # # All configuration values have a default; values that are commented out # serve to show the default. # If extensions (or modules to document with autodoc) are in another directory, # add these directories to sys.path here. If the directory is relative to the # documentation root, use os.path.abspath to make it absolute, like shown here. # # import os # import sys # sys.path.insert(0, os.path.abspath('.')) from iss.version import __version__ # -- General configuration ------------------------------------------------ # If your documentation needs a minimal Sphinx version, state it here. # # needs_sphinx = '1.0' # Add any Sphinx extension module names here, as strings. They can be # extensions coming with Sphinx (named 'sphinx.ext.*') or your custom # ones. extensions = [ 'sphinx.ext.autodoc', 'sphinx.ext.napoleon', 'sphinx.ext.viewcode'] # Add any paths that contain templates here, relative to this directory. templates_path = ['_templates'] # The suffix(es) of source filenames. # You can specify multiple suffix as a list of string: # # source_suffix = ['.rst', '.md'] source_suffix = '.rst' # The master toctree document. master_doc = 'index' # General information about the project. project = 'InSilicoSeq' copyright = '2017, Hadrien Gourle' author = 'Hadrien Gourle' # The version info for the project you're documenting, acts as replacement for # |version| and |release|, also used in various other places throughout the # built documents. # # The short X.Y version. version = __version__[:-2] # The full version, including alpha/beta/rc tags. release = __version__ # The language for content autogenerated by Sphinx. Refer to documentation # for a list of supported languages. # # This is also used if you do content translation via gettext catalogs. # Usually you set "language" from the command line for these cases. language = None # List of patterns, relative to source directory, that match files and # directories to ignore when looking for source files. # This patterns also effect to html_static_path and html_extra_path exclude_patterns = ['_build', 'Thumbs.db', '.DS_Store'] # The name of the Pygments (syntax highlighting) style to use. pygments_style = 'sphinx' # If true, `todo` and `todoList` produce output, else they produce nothing. todo_include_todos = False # -- Options for HTML output ---------------------------------------------- # The theme to use for HTML and HTML Help pages. See the documentation for # a list of builtin themes. # html_theme = 'alabaster' # Theme options are theme-specific and customize the look and feel of a theme # further. For a list of options available for each theme, see the # documentation. # # html_theme_options = {} # Add any paths that contain custom static files (such as style sheets) here, # relative to this directory. They are copied after the builtin static files, # so a file named "default.css" will overwrite the builtin "default.css". html_static_path = ['_static'] # -- Options for HTMLHelp output ------------------------------------------ # Output file base name for HTML help builder. htmlhelp_basename = 'InSilicoSeqdoc' # -- Options for LaTeX output --------------------------------------------- latex_elements = { # The paper size ('letterpaper' or 'a4paper'). # # 'papersize': 'letterpaper', # The font size ('10pt', '11pt' or '12pt'). # # 'pointsize': '10pt', # Additional stuff for the LaTeX preamble. # # 'preamble': '', # Latex figure (float) alignment # # 'figure_align': 'htbp', } # Grouping the document tree into LaTeX files. List of tuples # (source start file, target name, title, # author, documentclass [howto, manual, or own class]). latex_documents = [ (master_doc, 'InSilicoSeq.tex', 'InSilicoSeq Documentation', 'Hadrien Gourlé', 'manual'), ] # -- Options for manual page output --------------------------------------- # One entry per manual page. List of tuples # (source start file, name, description, authors, manual section). man_pages = [ (master_doc, 'insilicoseq', 'InSilicoSeq Documentation', [author], 1) ] # -- Options for Texinfo output ------------------------------------------- # Grouping the document tree into Texinfo files. List of tuples # (source start file, target name, title, author, # dir menu entry, description, category) texinfo_documents = [ (master_doc, 'InSilicoSeq', 'InSilicoSeq Documentation', author, 'InSilicoSeq', 'One line description of project.', 'Miscellaneous'), ] InSilicoSeq-1.5.4/doc/index.rst000066400000000000000000000025361412502470100163130ustar00rootroot00000000000000.. InSilicoSeq documentation master file, created by sphinx-quickstart on Tue May 30 11:45:01 2017. You can adapt this file completely to your liking, but it should at least contain the root `toctree` directive. InSilicoSeq ============ InSilicoSeq is a sequencing simulator producing realistic Illumina reads. Primarily intended for simulating metagenomic samples, it can also be used to produce sequencing data from a single genome. InSilicoSeq is written in python, and use kernel density estimators to model the read quality of real sequencing data. InSilicoSeq supports substitution, insertion, deletion errors, and models gc bias and insert size distribution. Details ------- * **Authors:** Hadrien Gourlé, Juliette Hayer, Oskar E. Karlsson and Erik Bongcam-Rudloff * **Contact:** `hadrien.gourle@slu.se `_ * **GitHub:** `HadrienG/InSilicoSeq `_ * **License:** MIT * **Article:** `10.1093/bioinformatics/bty630 `_ Contents -------- .. toctree:: :maxdepth: 2 :glob: iss/install iss/generate iss/model iss/iss Indices and tables ------------------ * :ref:`genindex` * :ref:`modindex` * :ref:`search` .. |date| date:: .. |time| date:: %H:%M *This documentation was generated on* |date| *at* |time|. 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To install InSilicoSeq, type the following in your terminal: .. code-block:: bash pip install InSilicoSeq It will install InSilicoSeq as well as the following dependencies: * biopython * joblib * numpy * pysam * scipy Other installation options ^^^^^^^^^^^^^^^^^^^^^^^^^^ * With conda: .. code-block:: bash conda install -c bioconda insilicoseq * Upgrading InSilicoSeq to the latest version: .. code-block:: bash pip install --upgrade InSilicoSeq * If you don't have administration rights on your machine: .. code-block:: bash pip3 install --user InSilicoSeq * if you wish to install InSilicoSeq at a custom location (i.e with a module system): .. code-block:: bash prefix="/path/to/install/prefix" pip install --install-option="--prefix=/$prefix" InSilicoSeq then add ``$prefix/bin`` to your ``PATH``, and ``$prefix/lib/python3.X/site-packages`` to your ``PYTHONPATH`` (replacing python3.X with your python version) .. _using_docker: Using docker ----------- If you wish to use InSilicoSeq using docker .. code-block:: bash docker pull hadrieng/insilicoseq:latest To use InSilicoSeq with docker, you need to provide a `volume` to the ``docker run`` command. Given with the ``-v`` option, the volume is your way of exchanging data (in this case, your input and output files) with the docker container. .. code-block:: bash docker run -v /Users/hadrien/data:/mnt/data -it --rm \ hadrieng/insilicoseq iss generate \ --genomes /mnt/data/genomes.fasta -m miseq \ -o /mnt/data/reads The above command will mount the local folder ``/Users/hadrien/data`` onto ``/mnt/data`` on the docker side. The output reads will be located in ``/Users/hadrien/data`` when InSilicoSeq has finished running. InSilicoSeq-1.5.4/doc/iss/iss.error_models.rst000066400000000000000000000012521412502470100212650ustar00rootroot00000000000000iss\.error\_models package ========================== Submodules ---------- iss\.error\_models\.basic module -------------------------------- .. automodule:: iss.error_models.basic :members: :undoc-members: :show-inheritance: iss\.error\_models\.cdf module ------------------------------ .. automodule:: iss.error_models.cdf :members: :undoc-members: :show-inheritance: iss\.error\_models\.kde module ------------------------------ .. automodule:: iss.error_models.kde :members: :undoc-members: :show-inheritance: Module contents --------------- .. automodule:: iss.error_models :members: :undoc-members: :show-inheritance: InSilicoSeq-1.5.4/doc/iss/iss.rst000066400000000000000000000017041412502470100165740ustar00rootroot00000000000000iss package =========== Subpackages ----------- .. toctree:: iss.error_models Submodules ---------- iss\.abundance module --------------------- .. automodule:: iss.abundance :members: :undoc-members: :show-inheritance: iss\.app module --------------- .. automodule:: iss.app :members: :undoc-members: :show-inheritance: iss\.bam module --------------- .. automodule:: iss.bam :members: :undoc-members: :show-inheritance: iss\.generator module --------------------- .. automodule:: iss.generator :members: :undoc-members: :show-inheritance: iss\.modeller module -------------------- .. automodule:: iss.modeller :members: :undoc-members: :show-inheritance: iss\.util module ---------------- .. automodule:: iss.util :members: :undoc-members: :show-inheritance: Module contents --------------- .. automodule:: iss :members: :undoc-members: :show-inheritance: InSilicoSeq-1.5.4/doc/iss/model.rst000066400000000000000000000044531412502470100171020ustar00rootroot00000000000000.. _model: Error models ============ Pre-built models ---------------- Available models ^^^^^^^^^^^^^^^^ +------------+-------------+ | Model name | Read length | +============+=============+ | MiSeq | 300 bp | +------------+-------------+ | HiSeq | 125 bp | +------------+-------------+ | NovaSeq | 150 bp | +------------+-------------+ .. figure:: qualities.png Average per base quality profiles for the prebuilt error models Qualities ^^^^^^^^^ The prebuilt models were built with the following commands: .. code-block:: bash megahit -1 reads_R1.fastq -2 reads_R2.fastq -o asm bowtie2-build asm/final.contigs.fa miseq_asm/final.contigs bowtie2 -x asm/final.contigs -1 reads_R1.fastq \ -2 reads_R2.fastq | samtools view -bS | samtools sort -o mapping.bam samtools index mapping.bam iss model -b mapping.bam -o name_of_model The MiSeq model was built from an ocean metagenomics project in Kenya (sample accession number ERR1912174). The HiSeq and NovaSeq models were built from human run obtained via basespace. Creating an Error Model ----------------------- If you do not wish to use the pre-computed error models provided with InSilicoSeq, it is possible to create your own. InSilicoSeq creates error models from .bam files. The input bam file should be a set of reads aligned against a reference genome or metagenome. Given you have two read files, `reads_R1.fastq.gz` and`reads_R2.fastq.gz`, and a reference metagenome `ref.fasta`: Align you reads against the reference ^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^ .. code-block:: bash bowtie2-build ref.fasta ref bowtie2 -x ref -1 reads_R1.fastq.gz \ -2 reads_R2.fastq.gz | samtools view -bS | samtools sort -o ref.bam samtools index ref.bam Build the model ^^^^^^^^^^^^^^^ .. code-block:: bash iss model -b ref.bam -o my_model which will create a `my_model.npz` file containing your newly built model Full list of options -------------------- --bam ^^^^^ aligned reads from which the model will be inferred (Required) --model ^^^^^^^ Error model to build. If not specified, using kernel density estimation (default: kde). Can be 'kde' or 'cdf' --output ^^^^^^^^ Output file path and prefix (Required) --quiet ^^^^^^^ Disable info logging --debug ^^^^^^^ Enable debug logging InSilicoSeq-1.5.4/doc/iss/modules.rst000066400000000000000000000001071412502470100174420ustar00rootroot00000000000000InSilicoSeq =========== .. toctree:: :maxdepth: 4 iss setup InSilicoSeq-1.5.4/doc/iss/qualities.png000066400000000000000000016621251412502470100177650ustar00rootroot00000000000000PNG  IHDR qyiCCPkCGColorSpaceGenericRGB8U]hU>sg#$Sl4t? % V46nI6"dΘ83OEP|1Ŀ (>/ % (>P苦;3ie|{g蹪X-2s=+WQ+]L6O w[C{_F qb Uvz?Zb1@/zcs>~if,ӈUSjF 1_Mjbuݠpamhmçϙ>a\+5%QKFkm}ۖ?ޚD\!~6,-7SثŜvķ5Z;[rmS5{yDyH}r9|-ăFAJjI.[/]mK 7KRDrYQO-Q||6 (0 MXd(@h2_f<:”_δ*d>e\c?~,7?& ك^2Iq2"y@g|U\ @IDATxE,9HňNL" b,zSQ3+*ʝN3AE0 1 ( $I,zz{wgh\SOVZr\~{2zhy뭷d…<Ύ{rqIjժPX8v=N;Aɺu*9#f̝;W^~e_ve=ܲe#ȸq㬿*U$M4C9DFQG! 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InSilicoSeq-1.5.4/iss/abundance.py000066400000000000000000000221021412502470100167640ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from Bio import SeqIO from scipy import stats import os import sys import logging import numpy as np def parse_abundance_file(abundance_file): """Parse an abundance or coverage file The abundance/coverage file is a flat file of the format "genome_idabundance" or "genome_idcoverage" Args: abundance_file (string): the path to the abundance file Returns: dict: genome_id as keys, abundance as values """ logger = logging.getLogger(__name__) abundance_dic = {} try: assert os.stat(abundance_file).st_size != 0 f = open(abundance_file, 'r') except (IOError, OSError) as e: logger.error('Failed to open file:%s' % e) sys.exit(1) except AssertionError as e: logger.error('File seems empty: %s' % abundance_file) sys.exit(1) else: with f: for line in f: try: genome_id = line.split()[0] abundance = float(line.split()[1]) except IndexError as e: logger.error('Failed to read file: %s' % e) sys.exit(1) else: abundance_dic[genome_id] = abundance logger.debug('Loaded abundance/coverage file: %s' % abundance_file) return abundance_dic def uniform(record_list): """Generate uniform abundance distribution from a number of records Args: record_list (list): a list of record.id Returns: dict: a dictionary with records as keys, abundance as values """ abundance_dic = {} n_records = len(record_list) for record in record_list: abundance_dic[record] = 1 / n_records return abundance_dic def halfnormal(record_list): """Generate scaled halfnormal abundance distribution from a number of records Args: record_list (list): a list of record.id Returns: dict: a dictionary with records as keys, abundance as values """ abundance_dic = {} n_records = len(record_list) dist = stats.halfnorm.rvs(loc=0.00, scale=1.00, size=n_records) dist_scaled = dist / sum(dist) for record, abundance in zip(record_list, dist_scaled): abundance_dic[record] = abundance return abundance_dic def exponential(record_list): """Generate scaled exponential abundance distribution from a number of records Args: record_list (list): a list of record.id Returns: dict: a dictionary with records as keys, abundance as values """ abundance_dic = {} n_records = len(record_list) dist = np.random.exponential(size=n_records) dist_scaled = dist / sum(dist) for record, abundance in zip(record_list, dist_scaled): abundance_dic[record] = abundance return abundance_dic def lognormal(record_list): """Generate scaled lognormal abundance distribution from a number of records Args: record_list (list): a list of record.id Returns: dict: a dictionary with records as keys, abundance as values """ abundance_dic = {} n_records = len(record_list) dist = np.random.lognormal(size=n_records) dist_scaled = dist / sum(dist) for record, abundance in zip(record_list, dist_scaled): abundance_dic[record] = abundance return abundance_dic def zero_inflated_lognormal(record_list): """Generate scaled zero-inflated lognormal abundance distribution from a number of records Args: record_list (list): a list of record.id Returns: dict: a dictionary with records as keys, abundance as values """ abundance_dic = {} n_records = len(record_list) zero_inflated = stats.bernoulli.rvs(p=0.2, size=n_records) dist = (1 - zero_inflated) * np.random.lognormal(size=n_records) dist_scaled = dist / sum(dist) for record, abundance in zip(record_list, dist_scaled): abundance_dic[record] = abundance return abundance_dic def to_coverage(total_n_reads, species_abundance, read_length, genome_size): """Calculate the coverage of a genome in a metagenome given its size and abundance Args: total_n_reads (int): total amount of reads in the dataset species_abundance (float): abundance of the species, between 0 and 1 read_length (int): length of the reads in the dataset genome_size (int): size of the genome Returns: float: coverage of the genome """ n_reads = total_n_reads * species_abundance coverage = (n_reads * read_length) / genome_size return coverage def coverage_scaling(total_n_reads, abundance_dic, genome_file, read_length): """Scale coverage distribution according to the n_reads parameter Args: total_n_reads (int): total amount of reads to simulate abundance_dic (dict): a dictionary with records as keys, coverage as values genome_file (str): path to input fasta file containing genomes read_length (int): length of the reads in the dataset Returns: dict: scaled coverage dictionary """ total_reads = 0 f = open(genome_file, 'r') # re-opens the file with f: fasta_file = SeqIO.parse(f, 'fasta') for record in fasta_file: try: species_coverage = abundance_dic[record.id] except KeyError as e: logger.error( 'Fasta record not found in abundance file: %s' % e) sys.exit(1) genome_size = len(record.seq) reads_g = species_coverage * genome_size / read_length / 2 total_reads += reads_g scale_factor = total_n_reads / total_reads for key in abundance_dic: abundance_dic[key] *= scale_factor return abundance_dic def to_file(abundance_dic, output, mode="abundance"): """Write the abundance dictionary to a file Args: abundance_dic (dict): the abundance dictionary output (str): the output file name """ logger = logging.getLogger(__name__) if mode == "abundance": output_abundance = output + '_abundance.txt' else: output_abundance = output + '_coverage.txt' try: f = open(output_abundance, 'w') except PermissionError as e: logger.error('Failed to open output file: %s' % e) sys.exit(1) else: with f: for record, abundance in abundance_dic.items(): f.write('%s\t%s\n' % (record, abundance)) def draft(genomes, draft, distribution, output, mode="abundance"): """Computes the abundance dictionary for a mix of complete and draft genomes Args: genomes (list): list of all input records draft (list): draft genome files distribution (function): distribution function to use output (str): output file Returns: dict: the abundance dictionary """ # first we get a list of contig names in draft genomes draft_records = [] for d in draft: draft_records.extend( [record.name for record in SeqIO.parse(d, 'fasta')]) genomes = list(set(genomes) - set(draft_records)) abundance_dic = distribution(genomes + draft) complete_genomes_abundance = {k: v for k, v in abundance_dic.items() if k not in draft} to_file(abundance_dic, output) draft_dic = expand_draft_abundance(abundance_dic, draft, mode) full_abundance_dic = {**complete_genomes_abundance, **draft_dic} return full_abundance_dic def expand_draft_abundance(abundance_dic, draft, mode="abundance"): """Calculate abundance for each contig of a draft genome The function takes the abundance dictionary and automatically detects draft genomes. In coverage mode the function simply assign the coverage value to each contig Args: abundance_dic (dict): dict with genome (paths or id) as key and abundance as value draft (list): draft genome files mode (str): abundance or coverage Returns: dict: abundance dictionary with abundance value for each contig """ draft_dic = {} for key, abundance in abundance_dic.items(): if key in draft: try: f = open(key, 'r') with f: fasta_file = SeqIO.parse(f, 'fasta') draft_records = [record for record in fasta_file] total_length = sum(len(record) for record in draft_records) except Exception as e: raise else: total_length = sum(len(record) for record in draft_records) for record in draft_records: if mode == "abundance": length = len(record) contig_abundance = abundance * (length / total_length) # print(key, record.id, contig_abundance) draft_dic[record.id] = contig_abundance elif mode == "coverage": draft_dic[record.id] = abundance return draft_dic InSilicoSeq-1.5.4/iss/app.py000066400000000000000000000560671412502470100156450ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import bam from iss import util from iss import download from iss import abundance from iss import generator from iss.version import __version__ from Bio import SeqIO from joblib import Parallel, delayed, load, dump import gc import os import sys import pickle import random import logging import argparse import numpy as np def generate_reads(args): """Main function for the `iss generate` submodule This submodule generates reads from an ErrorModel and write them to args.output + _R(1|2).fastq Args: args (object): the command-line arguments from argparse """ logger = logging.getLogger(__name__) logger.debug('iss version %s' % __version__) logger.debug('Using verbose logger') try: # try to import and load the correct error model logger.info('Starting iss generate') logger.info('Using %s ErrorModel' % args.mode) if args.seed: logger.info('Setting random seed to %i' % args.seed) random.seed(args.seed) np.random.seed(args.seed) if args.mode == 'kde': from iss.error_models import kde if args.model is None: logger.error('--model is required in --mode kde') sys.exit(1) elif args.model.lower() == 'hiseq': npz = os.path.join( os.path.dirname(__file__), 'profiles/HiSeq') elif args.model.lower() == 'novaseq': npz = os.path.join( os.path.dirname(__file__), 'profiles/NovaSeq') elif args.model.lower() == 'miseq': npz = os.path.join( os.path.dirname(__file__), 'profiles/MiSeq') else: npz = args.model err_mod = kde.KDErrorModel(npz) elif args.mode == 'basic': if args.model is not None: logger.warning('--model %s will be ignored in --mode %s' % ( args.model, args.mode)) from iss.error_models import basic err_mod = basic.BasicErrorModel() elif args.mode == 'perfect': if args.model is not None: logger.warning('--model %s will be ignored in --mode %s' % ( args.model, args.mode)) from iss.error_models import perfect err_mod = perfect.PerfectErrorModel() except ImportError as e: logger.error('Failed to import ErrorModel module: %s' % e) sys.exit(1) try: # try to read genomes and concatenate --genomes and --ncbi genomes if args.genomes or args.draft or args.ncbi: genome_files = [] if args.genomes: genome_files.extend(args.genomes) if args.draft: genome_files.extend(args.draft) if args.ncbi and args.n_genomes_ncbi: util.genome_file_exists(args.output + '_ncbi_genomes.fasta') total_genomes_ncbi = [] try: assert len(*args.ncbi) == len(*args.n_genomes_ncbi) except AssertionError as e: logger.error( '--ncbi and --n_genomes_ncbi of unequal lengths. \ Aborting') sys.exit(1) for g, n in zip(*args.ncbi, *args.n_genomes_ncbi): genomes_ncbi = download.ncbi( g, n, args.output + '_ncbi_genomes.fasta') genome_files.append(genomes_ncbi) if args.ncbi and not args.n_genomes_ncbi: logger.error( '--ncbi/-k requires --n_genomes_ncbi/-U. Aborting.') sys.exit(1) else: logger.error("One of --genomes/-g, --draft, --ncbi/-k is required") sys.exit(1) genome_file = args.output + '.iss.tmp.genomes.fasta' util.concatenate( genome_files, output=genome_file) # for n_genomes we use reservoir sampling to draw random genomes # from the concatenated genome file. We then override the file. if args.n_genomes and not args.draft and not args.ncbi: genome_count = util.count_records(genome_file) genome_files = [genome for genome in util.reservoir( SeqIO.parse(genome_file, 'fasta'), genome_count, args.n_genomes)] SeqIO.write(genome_files, genome_file, 'fasta') assert os.stat(genome_file).st_size != 0 f = open(genome_file, 'r') with f: # count the number of records genome_list = util.count_records(f) except IOError as e: logger.error('Failed to open genome(s) file:%s' % e) sys.exit(1) except AssertionError as e: logger.error('Genome(s) file seems empty: %s' % genome_file) sys.exit(1) except KeyboardInterrupt as e: logger.error('iss generate interrupted: %s' % e) sys.exit(1) else: abundance_dispatch = { 'uniform': abundance.uniform, 'halfnormal': abundance.halfnormal, 'exponential': abundance.exponential, 'lognormal': abundance.lognormal, 'zero_inflated_lognormal': abundance.zero_inflated_lognormal } # read the abundance file if args.abundance_file: logger.info('Using abundance file:%s' % args.abundance_file) if args.draft: abundance_dic_short = abundance.parse_abundance_file( args.abundance_file) complete_genomes_dic = {k: v for k, v in abundance_dic_short.items() if k not in args.draft} draft_dic = abundance.expand_draft_abundance( abundance_dic_short, args.draft) abundance_dic = {**complete_genomes_dic, **draft_dic} else: abundance_dic = abundance.parse_abundance_file( args.abundance_file) elif args.coverage_file: logger.warning('--coverage_file is an experimental feature') logger.warning('--coverage_file disables --n_reads') logger.info('Using coverage file:%s' % args.coverage_file) if args.draft: coverage_dic = abundance.parse_abundance_file( args.coverage_file) complete_genomes_dic = {k: v for k, v in coverage_dic.items() if k not in args.draft} draft_dic = abundance.expand_draft_abundance( abundance_dic_short, args.draft, mode="coverage") abundance_dic = {**complete_genomes_dic, **draft_dic} else: abundance_dic = abundance.parse_abundance_file( args.coverage_file) elif args.coverage in abundance_dispatch: # todo coverage distribution with --draft logger.warning('--coverage is an experimental feature') logger.info('Using %s coverage distribution' % args.coverage) if args.draft: abundance_dic = abundance.draft( genome_list, args.draft, abundance_dispatch[args.abundance], args.output, mode="coverage") else: abundance_dic = abundance_dispatch[ args.coverage](genome_list) if args.n_reads: n_reads = util.convert_n_reads(args.n_reads) logger.info('scaling coverage to %s reads' % n_reads) abundance_dic = abundance.coverage_scaling(n_reads, abundance_dic, genome_file, err_mod.read_length) abundance.to_file(abundance_dic, args.output, mode="coverage") elif args.abundance in abundance_dispatch: logger.info('Using %s abundance distribution' % args.abundance) if args.draft: abundance_dic = abundance.draft( genome_list, args.draft, abundance_dispatch[args.abundance], args.output) else: abundance_dic = abundance_dispatch[ args.abundance](genome_list) abundance.to_file(abundance_dic, args.output) else: logger.error('Could not get abundance') sys.exit(1) cpus = args.cpus logger.info('Using %s cpus for read generation' % cpus) if not (args.coverage or args.coverage_file): n_reads = util.convert_n_reads(args.n_reads) logger.info('Generating %s reads' % n_reads) try: temp_file_list = [] # list holding the prefix of all temp files f = open(genome_file, 'r') # re-opens the file with f: fasta_file = SeqIO.parse(f, 'fasta') total_reads_generated = 0 total_reads_generated_unrouned = 0 for record in fasta_file: # generate reads for records try: species_abundance = abundance_dic[record.id] except KeyError as e: logger.error( 'Fasta record not found in abundance file: %s' % e) sys.exit(1) else: logger.info('Generating reads for record: %s' % record.id) genome_size = len(record.seq) if args.coverage or args.coverage_file: coverage = species_abundance else: coverage = abundance.to_coverage( n_reads, species_abundance, err_mod.read_length, genome_size ) n_pairs_unrounded = ( (coverage * len(record.seq)) / err_mod.read_length) / 2 n_pairs = round(n_pairs_unrounded) # check that the rounding does not cause to drop # read pairs total_reads_generated_unrouned += n_pairs_unrounded total_reads_generated += n_pairs if round(total_reads_generated_unrouned) > \ total_reads_generated: logger.debug( "Adding a pair to correct rounding error") n_pairs += 1 total_reads_generated += 1 # skip record if n_reads == 0 if n_pairs == 0: continue # exact n_reads for each cpus if n_pairs % cpus == 0: n_pairs_per_cpu = [(n_pairs // cpus) for _ in range(cpus)] else: n_pairs_per_cpu = [(n_pairs // cpus) for _ in range(cpus)] n_pairs_per_cpu[-1] += n_pairs % cpus # due to a bug in multiprocessing # https://bugs.python.org/issue17560 # we can't send records taking more than 2**31 bytes # through serialisation. # In those cases we use memmapping if sys.getsizeof(str(record.seq)) >= 2**31 - 1: logger.warning( "record %s unusually big." % record.id) logger.warning("Using a memory map.") mode = "memmap" record_mmap = "%s.memmap" % args.output if os.path.exists(record_mmap): os.unlink(record_mmap) util.dump(record, record_mmap) del record record = record_mmap gc.collect() else: mode = "default" record_file_name_list = Parallel( n_jobs=cpus)( delayed(generator.reads)( record, err_mod, n_pairs_per_cpu[i], i, args.output, args.seed, args.gc_bias, mode) for i in range(cpus)) temp_file_list.extend(record_file_name_list) except KeyboardInterrupt as e: logger.error('iss generate interrupted: %s' % e) temp_file_unique = list(set(temp_file_list)) temp_R1 = [temp_file + '_R1.fastq' for temp_file in temp_file_list] temp_R2 = [temp_file + '_R2.fastq' for temp_file in temp_file_list] full_tmp_list = temp_R1 + temp_R2 full_tmp_list.append(genome_file) if os.path.exists("%s.memmap" % args.output): full_tmp_list.append("%s.memmap" % args.output) util.cleanup(full_tmp_list) sys.exit(1) else: # remove the duplicates in file list and cleanup # we remove the duplicates in case two records had the same header # and reads were appended to the same temp file. temp_file_unique = list(set(temp_file_list)) temp_R1 = [temp_file + '_R1.fastq' for temp_file in temp_file_list] temp_R2 = [temp_file + '_R2.fastq' for temp_file in temp_file_list] util.concatenate(temp_R1, args.output + '_R1.fastq') util.concatenate(temp_R2, args.output + '_R2.fastq') full_tmp_list = temp_R1 + temp_R2 full_tmp_list.append(genome_file) if os.path.exists("%s.memmap" % args.output): full_tmp_list.append("%s.memmap" % args.output) util.cleanup(full_tmp_list) if args.compress: util.compress(args.output + '_R1.fastq') util.compress(args.output + '_R2.fastq') logger.info('Read generation complete') def model_from_bam(args): """Main function for the `iss model` submodule This submodule write all variables necessary for building an ErrorModel to args.output + .npz Args: args (object): the command-line arguments from argparse """ logger = logging.getLogger(__name__) logger.debug('iss version %s' % __version__) logger.debug('Using verbose logger') try: # try to import bam module and write model data to file logger.info('Starting iss model') from iss import bam except ImportError as e: logger.error('Failed to import bam module: %s' % e) sys.exit(1) else: logger.info('Using KDE ErrorModel') bam.to_model(args.bam, args.output) logger.info('Model generation complete') def main(): parser = argparse.ArgumentParser( prog='InSilicoSeq', usage='iss [subcommand] [options]', description='InSilicoSeq: A sequencing simulator' ) parser.add_argument( '-v', '--version', action='store_true', default=False, help='print software version and exit' ) subparsers = parser.add_subparsers( title='available subcommands', metavar='' ) parser_mod = subparsers.add_parser( 'model', prog='iss model', description='generate an error model from a bam file', help='generate an error model from a bam file' ) parser_gen = subparsers.add_parser( 'generate', prog='iss generate', description='simulate reads from an error model', help='simulate reads from an error model' ) # arguments form the read generator module param_logging = parser_gen.add_mutually_exclusive_group() # --genomes and --ncbi should not be exclusive anymore # input_genomes = parser_gen.add_mutually_exclusive_group() input_abundance = parser_gen.add_mutually_exclusive_group() param_logging.add_argument( '--quiet', '-q', action='store_true', default=False, help='Disable info logging. (default: %(default)s).' ) param_logging.add_argument( '--debug', '-d', action='store_true', default=False, help='Enable debug logging. (default: %(default)s).' ) parser_gen.add_argument( '--seed', type=int, metavar='', help='Seed all the random number generators', default=None ) parser_gen.add_argument( '--cpus', '-p', default=2, type=int, metavar='', help='number of cpus to use. (default: %(default)s).' ) parser_gen.add_argument( '--genomes', '-g', metavar='', nargs="+", help='Input genome(s) from where the reads will originate' ) parser_gen.add_argument( '--draft', metavar='', nargs="+", help='Input draft genome(s) from where the reads will originate' ) parser_gen.add_argument( '--n_genomes', '-u', type=int, metavar='', help='How many genomes will be used for the simulation. is set with \ --genomes/-g or/and --draft to take random genomes from the \ input multifasta' ) parser_gen.add_argument( '--ncbi', '-k', choices=['bacteria', 'viruses', 'archaea'], action='append', nargs='*', metavar='', help='Download input genomes from NCBI. Requires --n_genomes/-u\ option. Can be bacteria, viruses, archaea or a combination of the\ three (space-separated)' ) parser_gen.add_argument( '--n_genomes_ncbi', '-U', type=int, action='append', metavar='', nargs='*', help='How many genomes will be downloaded from NCBI. Required if\ --ncbi/-k is set. If more than one kingdom is set with --ncbi,\ multiple values are necessary (space-separated).' ) input_abundance.add_argument( '--abundance', '-a', choices=['uniform', 'halfnormal', 'exponential', 'lognormal', 'zero_inflated_lognormal'], metavar='', default='lognormal', help='abundance distribution (default: %(default)s). Can be uniform,\ halfnormal, exponential, lognormal or zero-inflated-lognormal.' ) input_abundance.add_argument( '--abundance_file', '-b', metavar='', help='abundance file for coverage calculations (default: %(default)s).' ) input_abundance.add_argument( '--coverage', '-C', choices=['uniform', 'halfnormal', 'exponential', 'lognormal', 'zero_inflated_lognormal'], metavar='', help='coverage distribution. Can be uniform,\ halfnormal, exponential, lognormal or zero-inflated-lognormal.' ) input_abundance.add_argument( '--coverage_file', '-D', metavar='', help='file containing coverage information (default: %(default)s).' ) parser_gen.add_argument( '--n_reads', '-n', metavar='', default='1000000', help='Number of reads to generate (default: %(default)s). Allows \ suffixes k, K, m, M, g and G (ex 0.5M for 500000).' ) parser_gen.add_argument( '--mode', '-e', metavar='', choices=['kde', 'basic', 'perfect'], default='kde', help='Error model. If not specified, using kernel density estimation \ (default: %(default)s). Can be kde, basic or perfect' ) parser_gen.add_argument( '--model', '-m', metavar='', default=None, help='Error model file. (default: %(default)s). Use HiSeq, NovaSeq or \ MiSeq for a pre-computed error model provided with the software, or a \ file generated with iss model. If you do not wish to use a model, use \ --mode basic or --mode perfect. The name of the built-in models are \ case insensitive.' ) parser_gen.add_argument( '--gc_bias', '-c', action='store_true', default=False, help='If set, may fail to sequence reads with abnormal GC content. \ (default: %(default)s)' ) parser_gen.add_argument( '--compress', '-z', action='store_true', default=False, help='Compress the output in gzip format (default: %(default)s).' ) parser_gen.add_argument( '--output', '-o', metavar='', help='Output file path and prefix (Required)', required=True ) parser_gen._optionals.title = 'arguments' parser_gen.set_defaults(func=generate_reads) # arguments for the error model module parser_mod.add_argument( '--quiet', '-q', action='store_true', default=False, help='Disable info logging. (default: %(default)s).' ) parser_mod.add_argument( '--debug', '-d', action='store_true', default=False, help='Enable debug logging. (default: %(default)s).' ) parser_mod.add_argument( '--bam', '-b', metavar='', help='aligned reads from which the model will be inferred (Required)', required=True ) parser_mod.add_argument( '--output', '-o', metavar='', help='Output file path and prefix (Required)', required=True ) parser_mod._optionals.title = 'arguments' parser_mod.set_defaults(func=model_from_bam) args = parser.parse_args() # set logger and display version if args.version try: if args.version: print('iss version %s' % __version__) sys.exit(0) elif args.quiet: logging.basicConfig(level=logging.ERROR) elif args.debug: logging.basicConfig(level=logging.DEBUG) else: logging.basicConfig(level=logging.INFO) args.func(args) logging.shutdown() except AttributeError as e: logger = logging.getLogger(__name__) logger.debug(e) parser.print_help() # raise # extra traceback to uncomment if all hell breaks lose InSilicoSeq-1.5.4/iss/bam.py000066400000000000000000000217031412502470100156110ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util from iss import modeller from scipy import stats from random import random import sys import pysam import logging import numpy as np def read_bam(bam_file, n_reads=1000000): """Bam file reader. Select random mapped reads from a bam file Args: bam_file (string): path to a bam file Yields: read: a pysam read object """ logger = logging.getLogger(__name__) try: lines = pysam.idxstats(bam_file).splitlines() total_records = sum([int(l.split("\t")[2]) for l in lines if not l.startswith("#")]) # total_records = sum(1 for _ in bam.fetch() if not _.is_unmapped) random_fraction = n_reads / total_records bam = pysam.AlignmentFile(bam_file, 'rb') # reopen the file except (IOError, ValueError, ZeroDivisionError, pysam.utils.SamtoolsError) as e: logger.error('Failed to read bam file: %s' % e) sys.exit(1) else: logger.info('Reading bam file: %s' % bam_file) c = 0 with bam: for read in bam.fetch(): if not read.is_unmapped and random() < random_fraction: c += 1 if logger.getEffectiveLevel() == 10: print( 'DEBUG:iss.bam:Subsampling %s / %s reads' % ( c, n_reads), end='\r') yield read elif c >= n_reads: break def write_to_file(model, read_length, mean_f, mean_r, hist_f, hist_r, sub_f, sub_r, ins_f, ins_r, del_f, del_r, i_size, output): """Write variables to a .npz file Args: model (string): the type of error model read_length (int): read length of the dataset mean_f (list): list of mean bin sizes mean_r (list): list of mean bin sizes hist_f (list): list of cumulative distribution functions for the forward read quality hist_r (list): list of cumulative distribution functions for the reverse read quality sub_f (list): list of dictionaries representing the substitution probabilities for the forward reads sub_r (list): list of dictionaries representing the substitution probabilities for the reverse reads ins_f (list): list of dictionaries representing the insertion probabilities for the forward reads ins_r (list): list of dictionaries representing the insertion probabilities for the reverse reads del_f (list): list of dictionaries representing the deletion probabilities for the forward reads del_r (list): list of dictionaries representing the deletion probabilities for the reverse reads i_size (int): distribution of insert size for the aligned reads output (string): prefix of the output file """ logger = logging.getLogger(__name__) try: logger.info('Writing model to file: %s' % output) np.savez_compressed( output, model=model, read_length=read_length, insert_size=i_size, mean_count_forward=mean_f, mean_count_reverse=mean_r, quality_hist_forward=hist_f, quality_hist_reverse=hist_r, subst_choices_forward=sub_f, subst_choices_reverse=sub_r, ins_forward=ins_f, ins_reverse=ins_r, del_forward=del_f, del_reverse=del_r ) except PermissionError as e: logger.error('Failed to open output file: %s' % e) sys.exit(1) def to_model(bam_path, output): """from a bam file, write all variables needed for modelling reads in a .npz model file For a brief description of the variables that will be written to the output file, see the bam.write_to_file function Args: bam_path (string): path to a bam file output (string): prefix of the output file """ logger = logging.getLogger(__name__) insert_size_dist = [] qualities_forward = [] qualities_reverse = [] subst_matrix_f = np.zeros([301, 16]) # we dont know the len of the reads subst_matrix_r = np.zeros([301, 16]) # yet. we will find out from the indel_matrix_f = np.zeros([301, 9]) # len of the quality lists indel_matrix_r = np.zeros([301, 9]) # read the bam file and extract info needed for modelling for read in read_bam(bam_path): # get insert size distribution if read.is_proper_pair: template_length = abs(read.template_length) i_size = template_length - (2 * len(read.seq)) insert_size_dist.append(i_size) # get qualities if read.is_read1: # get mean quality too quality_means = [] read_quality = read.query_qualities mean_quality = np.mean(read_quality) if read.is_reverse: read_quality = read_quality[::-1] # reverse the list quality_plus_mean = [ (quality, mean_quality) for quality in read_quality] qualities_forward.append(np.asarray(quality_plus_mean)) # qualities_forward.append(read.query_qualities) elif read.is_read2: # get mean quality too quality_means = [] read_quality = read.query_qualities mean_quality = np.mean(read_quality) if read.is_reverse: read_quality = read_quality[::-1] # reverse the list quality_plus_mean = [ (quality, mean_quality) for quality in read_quality] qualities_reverse.append(np.asarray(quality_plus_mean)) # qualities_reverse.append(read.query_qualities) # get mismatches alignment = read.get_aligned_pairs( matches_only=True, with_seq=True ) read_has_indels = False for base in alignment: # dispatch mismatches in matrix pos, subst, read_has_indels = modeller.dispatch_subst( base, read, read_has_indels) if read.is_read1 and subst is not None: subst_matrix_f[pos, subst] += 1 elif read.is_read2 and subst is not None: subst_matrix_r[pos, subst] += 1 if read_has_indels: # dispatch indels in matrix for pos, indel in modeller.dispatch_indels(read): if read.is_read1: indel_matrix_f[pos, indel] += 1 elif read.is_read2: indel_matrix_r[pos, indel] += 1 logger.info('Calculating insert size distribution') # insert_size = int(np.mean(insert_size_dist)) hist_insert_size = modeller.insert_size(insert_size_dist) logger.info('Calculating mean and base quality distribution') quality_bins_f = modeller.divide_qualities_into_bins(qualities_forward) quality_bins_r = modeller.divide_qualities_into_bins(qualities_reverse) # getting distribution of mean sequence quality mean_f = [len(quality_bin) for quality_bin in quality_bins_f] mean_r = [len(quality_bin) for quality_bin in quality_bins_r] hists_f = modeller.quality_bins_to_histogram(quality_bins_f) hists_r = modeller.quality_bins_to_histogram(quality_bins_r) # modern illumina instruments return reads of the same length # in case our bam file contains aligned reads of different length, # we coerce the model's read length to the smallest read of the bam file length_forward = min((len(x) for x in hists_f if len(x) > 1)) length_reverse = min((len(x) for x in hists_r if len(x) > 1)) read_length = min(length_forward, length_reverse) # now we can resize the substitution and indel matrices before # doing operations on them subst_matrix_f.resize([read_length, 16], refcheck=False) subst_matrix_r.resize([read_length, 16], refcheck=False) indel_matrix_f.resize([read_length, 9], refcheck=False) indel_matrix_r.resize([read_length, 9], refcheck=False) logger.info('Calculating substitution rate') subst_f = modeller.subst_matrix_to_choices(subst_matrix_f, read_length) subst_r = modeller.subst_matrix_to_choices(subst_matrix_r, read_length) logger.info('Calculating indel rate') # update the base count in indel matrices for position in range(read_length): indel_matrix_f[position][0] = sum(subst_matrix_f[position][::4]) indel_matrix_r[position][0] = sum(subst_matrix_r[position][::4]) ins_f, del_f = modeller.indel_matrix_to_choices( indel_matrix_f, read_length) ins_r, del_r = modeller.indel_matrix_to_choices( indel_matrix_r, read_length) write_to_file( 'kde', read_length, mean_f, mean_r, hists_f, hists_r, subst_f, subst_r, ins_f, ins_r, del_f, del_r, hist_insert_size, output + '.npz') InSilicoSeq-1.5.4/iss/download.py000066400000000000000000000072061412502470100166630ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from Bio import SeqIO from Bio import Entrez import io import sys import time import zlib import random import logging import requests class BadRequestError(Exception): """Exception to raise when a http request does not return 200 """ def __init__(self, url, status_code): super().__init__('%s returned %d' % (url, status_code)) def ncbi(kingdom, n_genomes, output): """download random genomes sequences from ncbi genomes with entrez eutils and requests. Args: kingdom (string): the kingdom from which the sequences are from n_genomes (int): the number of genomes to download Returns: str: the output file """ logger = logging.getLogger(__name__) Entrez.email = 'hadrien.gourle@slu.se' Entrez.tool = 'InSilicoSeq' Entrez.api_key = 'd784b36672ca73601f4a19c3865775a17207' full_id_list = Entrez.read(Entrez.esearch( 'assembly', term='%s[Organism] AND "latest refseq"[filter] AND "complete genome"[filter]' % kingdom, retmax=100000))['IdList'] n = 0 logger.info('Searching for %s to download' % kingdom) while n < n_genomes: ident = random.choice(full_id_list) genome_info = Entrez.read( Entrez.esummary( db='assembly', id=ident), validate=False)["DocumentSummarySet"]["DocumentSummary"][0] if genome_info['FtpPath_RefSeq']: url = genome_info['FtpPath_RefSeq'] url = "%s/%s_%s_genomic.fna.gz" \ % (genome_info['FtpPath_RefSeq'], genome_info['AssemblyAccession'], genome_info['AssemblyName']) logger.info('Downloading %s' % genome_info['AssemblyAccession']) try: assembly_to_fasta(url, output) except BadRequestError as e: logger.debug('Could not download %s' % genome_info['AssemblyAccession']) logger.debug('Skipping and waiting two seconds') time.sleep(2) else: n += 1 full_id_list.remove(ident) return output def assembly_to_fasta(url, output, chunk_size=1024): """download an assembly from the ncbi ftp and append the chromosome sequence to a fasta file. This function discards the plsamid sequences! Args: url (string): an url to a fasta file output (string): the output file name Returns: str: the file name """ logger = logging.getLogger(__name__) if url.startswith("ftp://"): # requests doesnt support ftp url = url.replace("ftp://", "https://") if url: request = requests.get(url) if request.status_code == 200: request = zlib.decompress( request.content, zlib.MAX_WBITS | 32).decode() else: raise BadRequestError(url, request.status_code) with io.StringIO(request) as fasta_io: seq_handle = SeqIO.parse(fasta_io, 'fasta') chromosome = filter_plasmids(seq_handle) try: f = open(output, 'a') except (IOError, OSError) as e: logger.error('Failed to open output file: %s' % e) sys.exit(1) else: logger.debug('Writing genome to %s' % output) with f: SeqIO.write(chromosome, f, 'fasta') return output def filter_plasmids(handle): """returns the largest sequence from a sequence handle """ n = 0 for record in handle: if len(record) > n: n = len(record) largest = record return largest InSilicoSeq-1.5.4/iss/error_models/000077500000000000000000000000001412502470100171715ustar00rootroot00000000000000InSilicoSeq-1.5.4/iss/error_models/__init__.py000066400000000000000000000161251412502470100213070ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util import sys import random import logging import numpy as np class ErrorModel(object): """Main ErrorModel Class This class is used to create inheriting classes and contains all the functions that are shared by all ErrorModel classes """ @property def logger(self): component = "{}.{}".format(type(self).__module__, type(self).__name__) return logging.getLogger(component) def load_npz(self, npz_path, model): """load the error profile .npz file Args: npz_path (string): path to the npz file model (string): type of model. Could be 'cdf' or 'kde'. 'cdf' has been deprecated and is no longer available Returns: ndarray: numpy object containg variables necessary for error model construction """ try: error_profile = np.load(npz_path, allow_pickle=True) assert error_profile['model'] == model except (OSError, IOError) as e: self.logger.error('Failed to read ErrorModel file: %s' % e) sys.exit(1) except AssertionError as e: self.logger.error( 'Trying to load a %s ErrorModel in %s mode' % ( error_profile['model'], model)) sys.exit(1) else: self.logger.debug('Loaded ErrorProfile: %s' % npz_path) return error_profile def introduce_error_scores(self, record, orientation): """Add phred scores to a SeqRecord according to the error_model Args: record (SeqRecord): a read record orientation (string): orientation of the read. Can be 'forward' or 'reverse' Returns: SeqRecord: a read record with error scores """ if orientation == 'forward': record.letter_annotations["phred_quality"] = self.gen_phred_scores( self.quality_forward, 'forward') elif orientation == 'reverse': record.letter_annotations["phred_quality"] = self.gen_phred_scores( self.quality_reverse, 'reverse') return record def mut_sequence(self, record, orientation): """Introduce substitution errors to a sequence If a random probability is higher than the probability of the basecall being correct, introduce a substitution error Args: record (SeqRecord): a read record with error scores orientation (string): orientation of the read. Can be 'forward' or 'reverse' Returns: Seq: a sequence """ # get the right subst_matrix if orientation == 'forward': nucl_choices = self.subst_choices_for elif orientation == 'reverse': nucl_choices = self.subst_choices_rev mutable_seq = record.seq.tomutable() quality_list = record.letter_annotations["phred_quality"] position = 0 for nucl, qual in zip(mutable_seq, quality_list): if random.random() > util.phred_to_prob(qual) \ and nucl.upper() not in 'RYWSMKHBVDN': mutable_seq[position] = str(np.random.choice( nucl_choices[position][nucl.upper()][0], p=nucl_choices[position][nucl.upper()][1])) position += 1 return mutable_seq.toseq() def adjust_seq_length(self, mut_seq, orientation, full_sequence, bounds): """Truncate or Extend reads to make them fit the read length When insertions or deletions are introduced to the reads, their length will change. This function takes a (mutable) read and a reference sequence, and extend or truncate the read if it has had an insertion or a deletion Args: mut_seq (MutableSeq): a mutable sequence orientation (string): orientation of the read. Can be 'forward' or 'reverse' full_sequence (Seq): the reference sequence from which mut_seq comes from bounds (tuple): the position of the read in the full_sequence Returns: Seq: a sequence fitting the ErrorModel """ read_start, read_end = bounds if len(mut_seq) == self.read_length: return mut_seq.toseq() elif len(mut_seq) > self.read_length: while len(mut_seq) > self.read_length: mut_seq.pop() return mut_seq.toseq() else: # len smaller to_add = self.read_length - len(mut_seq) if orientation == 'forward': for i in range(to_add): if read_end + i >= len(full_sequence): nucl_to_add = 'A' else: nucl_to_add = str(full_sequence[read_end + i]) mut_seq.append(nucl_to_add) elif orientation == 'reverse': for i in range(to_add): if read_end + i >= len(full_sequence): nucl_to_add = 'A' else: nucl_to_add = util.rev_comp( full_sequence[read_end + i]) mut_seq.append(nucl_to_add) return mut_seq.toseq() def introduce_indels(self, record, orientation, full_seq, bounds): """Introduce insertions or deletions in a sequence Introduce insertion and deletion errors according to the probabilities present in the indel choices list Args: record (SeqRecord): a sequence record orientation (string): orientation of the read. Can be 'forward' or 'reverse' full_seq (Seq): the reference sequence from which mut_seq comes from bounds (tuple): the position of the read in the full_sequence Returns: Seq: a sequence with (eventually) indels """ # get the right indel arrays if orientation == 'forward': insertions = self.ins_for deletions = self.del_for elif orientation == 'reverse': insertions = self.ins_rev deletions = self.del_rev mutable_seq = record.seq.tomutable() position = 0 for nucl in range(self.read_length - 1): try: # skip ambiguous nucleotides if mutable_seq[nucl].upper() in 'RYWSMKHBVDN': position += 1 continue for nucl_to_insert, prob in insertions[position].items(): if random.random() < prob: # we want to insert after the base read mutable_seq.insert(position + 1, str(nucl_to_insert)) if random.random() < deletions[position][mutable_seq[nucl].upper()]: mutable_seq.pop(position) position += 1 except IndexError as e: continue seq = self.adjust_seq_length( mutable_seq, orientation, full_seq, bounds) return seq InSilicoSeq-1.5.4/iss/error_models/basic.py000066400000000000000000000036411412502470100206300ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util from iss.error_models import ErrorModel from Bio.Seq import MutableSeq from Bio.SeqRecord import SeqRecord import random import numpy as np class BasicErrorModel(ErrorModel): """Basic Error Model class Basic error model. The phred scores are based on a normal distribution. Only substitutions errors occur. The substitution rate is assumed equal between all nucleotides. """ def __init__(self): super().__init__() self.read_length = 125 self.insert_size = 200 self.quality_forward = self.quality_reverse = 30 self.subst_choices_for = self.subst_choices_rev = [{ 'A': (['T', 'C', 'G'], [1/3, 1/3, 1/3]), 'T': (['A', 'C', 'G'], [1/3, 1/3, 1/3]), 'C': (['A', 'T', 'G'], [1/3, 1/3, 1/3]), 'G': (['A', 'T', 'C'], [1/3, 1/3, 1/3]) } for _ in range(self.read_length)] self.ins_for = self.ins_rev = self.del_for = self.del_rev = [{ 'A': 0.0, 'T': 0.0, 'C': 0.0, 'G': 0.0 } for _ in range(125)] def gen_phred_scores(self, mean_quality, orientation): """Generate a normal distribution, transform to phred scores Generate a list of phred score according to a normal distribution centered around the ErrorModel quality Args: mean_quality (int): mean phred score Returns: list: list of phred scores following a normal distribution """ norm = [min(q, 0.9999) for q in np.random.normal( util.phred_to_prob(mean_quality), 0.01, self.read_length)] phred = [util.prob_to_phred(p) for p in norm] return phred def random_insert_size(self): """Fake random function returning the default insert size of the basic arror model Returns: int: insert size """ return self.insert_size InSilicoSeq-1.5.4/iss/error_models/kde.py000066400000000000000000000065441412502470100203170ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util from iss.error_models import ErrorModel from Bio.Seq import MutableSeq from Bio.SeqRecord import SeqRecord from scipy import stats import sys import random import numpy as np class KDErrorModel(ErrorModel): """KDErrorModel class. Error model based on an .npz files derived from read alignments. the npz file must contain: - the length of the reads - the mean insert size - the size of mean sequence quality bins (for R1 and R2) - a cumulative distribution function of quality scores for each position (for R1 and R2) - the substitution for each nucleotide at each position (for R1 and R2) - the insertion and deletion rates for each position (for R1 and R2) """ def __init__(self, npz_path): super().__init__() self.npz_path = npz_path self.error_profile = self.load_npz(npz_path, 'kde') self.read_length = self.error_profile['read_length'] self.i_size_cdf = self.error_profile['insert_size'] self.mean_forward = self.error_profile['mean_count_forward'] self.mean_reverse = self.error_profile['mean_count_reverse'] self.quality_forward = self.error_profile['quality_hist_forward'] self.quality_reverse = self.error_profile['quality_hist_reverse'] self.subst_choices_for = self.error_profile['subst_choices_forward'] self.subst_choices_rev = self.error_profile['subst_choices_reverse'] self.ins_for = self.error_profile['ins_forward'] self.ins_rev = self.error_profile['ins_reverse'] self.del_for = self.error_profile['del_forward'] self.del_rev = self.error_profile['del_reverse'] self.error_profile = '' # discard the error profile after reading def gen_phred_scores(self, cdfs, orientation): """Generate a list of phred scores based on cdfs and mean bins For each position, draw a phred score from the cdf and append to the phred score list Args: cdfs (ndarray): array containing the cdfs orientation (string): orientation of the read. Can be 'forward' or 'reverse' Returns: list: a list of phred scores """ # choose which mean bin to use if orientation == 'forward': mean = self.mean_forward elif orientation == 'reverse': mean = self.mean_reverse norm_mean = mean / sum(mean) # quality_bin = np.searchsorted(norm_mean, np.random.rand()) quality_bin = np.random.choice(range(len(norm_mean)), p=norm_mean) # downgrades index out of bound (ex rand is 1, last bin in searchsorted # is 0.95) to best quality bin if quality_bin == 4: quality_bin = 3 cdfs_bin = cdfs[quality_bin] phred_list = [] for cdf in cdfs_bin: random_quality = np.searchsorted(cdf, np.random.rand()) phred_list.append(random_quality) return phred_list[:self.read_length] def random_insert_size(self): """Draw a random insert size from the insert size cdf Args: i_size_cdf: cumulative distribution function of the insert size Returns: int: an insert size """ insert_size = np.searchsorted(self.i_size_cdf, np.random.rand()) return insert_size InSilicoSeq-1.5.4/iss/error_models/perfect.py000066400000000000000000000027631412502470100212030ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util from iss.error_models import ErrorModel class PerfectErrorModel(ErrorModel): """Perfect error model class Perfect Error Model. This is a model without errors. All Phred score are 40. No errors are introduced at all. """ def __init__(self): super().__init__() self.read_length = 125 self.insert_size = 200 self.quality_forward = self.quality_reverse = 40 self.subst_choices_for = self.subst_choices_rev = [{ 'A': (['A', 'T', 'C', 'G'], [1, 0, 0, 0]), 'T': (['A', 'T', 'C', 'G'], [0, 1, 0, 0]), 'C': (['A', 'T', 'C', 'G'], [0, 0, 1, 0]), 'G': (['A', 'T', 'C', 'G'], [0, 0, 0, 1]) } for _ in range(self.read_length)] self.ins_for = self.ins_rev = self.del_for = self.del_rev = [{ 'A': 0.0, 'T': 0.0, 'C': 0.0, 'G': 0.0 } for _ in range(self.read_length)] def gen_phred_scores(self, mean_quality, orientation): """Fake randorm function returning the distribution of Phred scores. Score will be 40 for all positions Returns: list: list of phred scores (40 along the whole read) """ return [40 for _ in range(self.read_length)] def random_insert_size(self): """Fake random function returning the default insert size of the basic arror model Returns: int: insert size """ return self.insert_size InSilicoSeq-1.5.4/iss/generator.py000066400000000000000000000150011412502470100170320ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss.util import load, rev_comp from Bio import SeqIO from Bio.Seq import Seq from Bio.SeqUtils import GC from Bio.SeqRecord import SeqRecord from shutil import copyfileobj import os import sys import random import logging import numpy as np def reads(record, ErrorModel, n_pairs, cpu_number, output, seed, gc_bias=False, mode="default"): """Simulate reads from one genome (or sequence) according to an ErrorModel This function makes use of the `simulate_read` function to simulate reads and save them in a fastq file Args: record (SeqRecord): sequence or genome of reference ErrorModel (ErrorModel): an ErrorModel n_pairs (int): the number of reads to generate cpu_number (int): an int indentifying the cpu that is used by the function. Is used for naming the output file output (str): the output file prefix seed (int): random seed to use gc_bias (bool): if set, the function may skip a read due to abnormal GC content Returns: str: the name of the output file """ logger = logging.getLogger(__name__) # load the record from disk if mode is memmap if mode == "memmap": record_mmap = load(record) record = record_mmap if seed is not None: random.seed(seed + cpu_number) np.random.seed(seed + cpu_number) logger.debug( 'Cpu #%s: Generating %s read pairs' % (cpu_number, n_pairs)) read_tuple_list = [] i = 0 while i < n_pairs: # try: # forward, reverse = simulate_read(record, ErrorModel, i) # except ValueError as e: # logger.error('Skipping this record: %s' % record.id) # return try: forward, reverse = simulate_read(record, ErrorModel, i, cpu_number) except AssertionError as e: logger.warning( '%s shorter than read length for this ErrorModel' % record.id) logger.warning( 'Skipping %s. You will have less reads than specified' % record.id) break else: if gc_bias: stiched_seq = forward.seq + reverse.seq gc_content = GC(stiched_seq) if 40 < gc_content < 60: read_tuple_list.append((forward, reverse)) i += 1 elif np.random.rand() < 0.90: read_tuple_list.append((forward, reverse)) i += 1 else: continue else: read_tuple_list.append((forward, reverse)) i += 1 temp_file_name = output + '.iss.tmp.%s.%s' % (record.id, cpu_number) to_fastq(read_tuple_list, temp_file_name) return temp_file_name def simulate_read(record, ErrorModel, i, cpu_number): """From a read pair from one genome (or sequence) according to an ErrorModel Each read is a SeqRecord object returns a tuple containing the forward and reverse read. Args: record (SeqRecord): sequence or genome of reference ErrorModel (ErrorModel): an ErrorModel class i (int): a number identifying the read cpu_number (int): cpu number. Is added to the read id. Returns: tuple: tuple containg a forward read and a reverse read """ logger = logging.getLogger(__name__) sequence = record.seq header = record.id read_length = ErrorModel.read_length insert_size = ErrorModel.random_insert_size() # generate the forward read try: # a ref sequence has to be longer than 2 * read_length + i_size assert read_length < len(record.seq) forward_start = random.randrange( 0, len(record.seq) - (2 * read_length + insert_size)) except AssertionError as e: raise except ValueError as e: logger.debug( '%s shorter than template length for this ErrorModel:%s' % (record.id, e)) forward_start = max(0, random.randrange( 0, len(record.seq) - read_length)) # raise forward_end = forward_start + read_length bounds = (forward_start, forward_end) # create a perfect read forward = SeqRecord( Seq(str(sequence[forward_start:forward_end])), id='%s_%s_%s/1' % (header, i, cpu_number), description='' ) # add the indels, the qual scores and modify the record accordingly forward.seq = ErrorModel.introduce_indels( forward, 'forward', sequence, bounds) forward = ErrorModel.introduce_error_scores(forward, 'forward') forward.seq = ErrorModel.mut_sequence(forward, 'forward') # generate the reverse read try: reverse_start = forward_end + insert_size reverse_end = reverse_start + read_length assert reverse_end < len(record.seq) except AssertionError as e: # we use random insert when the modelled template length distribution # is too large reverse_end = random.randrange(read_length, len(record.seq)) reverse_start = reverse_end - read_length bounds = (reverse_start, reverse_end) # create a perfect read reverse = SeqRecord( Seq(rev_comp(str(sequence[reverse_start:reverse_end]))), id='%s_%s_%s/2' % (header, i, cpu_number), description='' ) # add the indels, the qual scores and modify the record accordingly reverse.seq = ErrorModel.introduce_indels( reverse, 'reverse', sequence, bounds) reverse = ErrorModel.introduce_error_scores(reverse, 'reverse') reverse.seq = ErrorModel.mut_sequence(reverse, 'reverse') return (forward, reverse) def to_fastq(generator, output): """Write reads to a fastq file Take a generator or a list containing read pairs (tuples) and write them in two fastq files: output_R1.fastq and output_R2.fastq Args: generator (generator): a read generator (or list) output (string): the output files prefix """ logger = logging.getLogger(__name__) # define name of output files output_forward = output + '_R1.fastq' output_reverse = output + '_R2.fastq' try: f = open(output_forward, 'a') r = open(output_reverse, 'a') except PermissionError as e: logger.error('Failed to open output file(s): %s' % e) sys.exit(1) else: with f, r: for read_tuple in generator: SeqIO.write(read_tuple[0], f, 'fastq-sanger') SeqIO.write(read_tuple[1], r, 'fastq-sanger') InSilicoSeq-1.5.4/iss/modeller.py000066400000000000000000000320041412502470100166510ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util from scipy import stats from joblib import Parallel, delayed import logging import numpy as np def insert_size(insert_size_distribution): """Calculate cumulative distribution function from the raw insert size distributin. Uses 1D kernel density estimation. Args: insert_size_distribution (list): list of insert sizes from aligned read pairs Returns: 1darray: a cumulative density function """ kde = stats.gaussian_kde( insert_size_distribution, bw_method=0.2 / np.std(insert_size_distribution, ddof=1)) x_grid = np.linspace( min(insert_size_distribution), max(insert_size_distribution), 1000) kde = kde.evaluate(x_grid) cdf = np.cumsum(kde) cdf = cdf / cdf[-1] return cdf def divide_qualities_into_bins(qualities, n_bins=4): """Divides the raw quality scores in bins according to the mean phred quality of the sequence they come from Args: qualities (list): raw count of all the phred scores and mean sequence quality n_bins (int): number of bins to create (default: 4) Returns: list: a list of lists containing the binned quality scores """ logger = logging.getLogger(__name__) logger.debug('Dividing qualities into mean clusters') bin_lists = [[] for _ in range(n_bins)] # create list of `n_bins` list ranges = np.split(np.array(range(40)), n_bins) for quality in qualities: mean = int(quality[0][1]) # mean is at 1 and same regardless of b pos which_array = 0 for array in ranges: if mean in array: read = np.fromiter((q[0] for q in quality), dtype=np.float) bin_lists[which_array].append(read) which_array += 1 return bin_lists def quality_bins_to_histogram(bin_lists): """Wrapper function to generate cdfs for each quality bins Generate cumulative distribution functions for a number of mean quality bins Args: bins_lists (list): list of list containing raw count of all phred scores Returns: list: a list of lists containg cumulative density functions """ logger = logging.getLogger(__name__) cdf_bins = [] i = 0 for qual_bin in bin_lists: if len(qual_bin) > 1: logger.debug('Transposing matrix for mean cluster #%s' % i) # quals = np.asarray(qual_bin).T # seems to make clunkier models quals = [q for q in zip(*qual_bin)] logger.debug( 'Modelling quality distribution for mean cluster #%s' % i) cdfs_list = raw_qualities_to_histogram(quals) cdf_bins.append(cdfs_list) else: logger.debug('Mean quality bin #%s of length < 1. Skipping' % i) cdf_bins.append([]) i += 1 return cdf_bins def raw_qualities_to_histogram(qualities): """Approximate the distribution of base quality at each position in a read using a pseudo 2d kernel density estimation Generate cumulative distribution functions Args: qualities (list): raw count of all phred scores Returns: list: list of cumulative distribution functions. One cdf per base. The list has the size of the read length """ logger = logging.getLogger(__name__) # moved this in quality_bins_to_histogram to try parallelization # quals = util.split_list([i for i in zip(*qualities)], n_parts=cpus) cdfs_list = [] for q in qualities: numpy_log_handler = np.seterrcall(util.nplog) with np.errstate(under='ignore', divide='call'): try: kde = stats.gaussian_kde(q, bw_method=0.2 / np.std(q, ddof=1)) except np.linalg.linalg.LinAlgError as e: # if np.std of array is 0, we modify the array slightly to be # able to divide by ~ np.std # this will print a FloatingPointError in DEBUG mode # logger.debug('np.std of quality array is zero: %s' % e) q = list(q) q[-1] += 1 kde = stats.gaussian_kde(q, bw_method=0.2 / np.std(q, ddof=1)) kde = kde.evaluate(range(41)) cdf = np.cumsum(kde) cdf = cdf / cdf[-1] cdfs_list.append(cdf) return cdfs_list def dispatch_subst(base, read, read_has_indels): """Return the x and y position of a substitution to be inserted in the substitution matrix. The substitution matrix is a 2D array of size 301 * 16 The x axis (301) corresponds to the position in the read, while the y axis (16) represents the match or substitution (see the dispatch dict in the function). Positions 0, 4, 8 and 12 are matches, other positions are substitutions The size of x axis is 301 because we haven't calculated the read length yet Args: base (tuple): one base from an aligmnent object. According to the pysam documentation: an alignment is a list of tuples: aligned read (query) and reference positions. the parameter with_seq adds the ref sequence as the 3rd element of the tuples. substitutions are lower-case. read (read): a read object, from which the alignment comes from read_has_indels (bool): a boolean flag to keep track if the read has an indel or not Returns: tuple: x and y position for incrementing the substitution matrix and a third element: True if an indel has been detected, False otherwise """ dispatch_dict = { 'AA': 0, 'aT': 1, 'aG': 2, 'aC': 3, 'TT': 4, 'tA': 5, 'tG': 6, 'tC': 7, 'CC': 8, 'cA': 9, 'cT': 10, 'cG': 11, 'GG': 12, 'gA': 13, 'gT': 14, 'gC': 15 } query_pos = base[0] query_base = read.seq[query_pos] ref_base = base[2] dispatch_key = ref_base + query_base if dispatch_key not in dispatch_dict: # flag reads that have one or more indels read_has_indels = True # flag the read for later indel treatment substitution = None # flag this base to skip substitution treatment else: substitution = dispatch_dict[dispatch_key] return (query_pos, substitution, read_has_indels) def subst_matrix_to_choices(substitution_matrix, read_length): """Transform a substitution matrix into probabilties of substitutions for each base and at every position From the raw mismatches at one position, returns a dictionary with probabilties of substitutions Args: substitution_matrix (np.array): the substitution matrix is a 2D array of size read_length * 16. fhe x axis (read_length) corresponds to the position in the read, while the y axis (16) represents the match or substitution. Positions 0, 4, 8 and 12 are matches, other positions are substitutions read_length (int): read length Returns: list: list of dictionaries representing the substitution probabilities for a collection of reads """ logger = logging.getLogger(__name__) nucl_choices_list = [] for pos in range(read_length): sums = { 'A': np.sum(substitution_matrix[pos][1:4]), 'T': np.sum(substitution_matrix[pos][5:8]), 'C': np.sum(substitution_matrix[pos][9:12]), 'G': np.sum(substitution_matrix[pos][13:]) } # we want to avoid 'na' in the data so we raise FloatingPointError # if we try to divide by 0 (no count data for that nucl at that pos) # we assume equal rate of substitution with np.errstate(all='raise'): nucl_choices = {} try: A = ( ['T', 'C', 'G'], [count / sums['A'] for count in substitution_matrix[pos][1:4]]) except FloatingPointError as e: logger.debug(e, exc_info=True) A = (['T', 'C', 'G'], [1/3, 1/3, 1/3]) try: T = ( ['A', 'C', 'G'], [count / sums['T'] for count in substitution_matrix[pos][5:8]]) except FloatingPointError as e: logger.debug(e, exc_info=True) T = (['A', 'C', 'G'], [1/3, 1/3, 1/3]) try: C = ( ['A', 'T', 'G'], [count / sums['C'] for count in substitution_matrix[pos][9:12]]) except FloatingPointError as e: logger.debug(e, exc_info=True) C = (['A', 'T', 'G'], [1/3, 1/3, 1/3]) try: G = ( ['A', 'T', 'C'], [count / sums['G'] for count in substitution_matrix[pos][13:]]) except FloatingPointError as e: logger.debug(e, exc_info=True) G = (['A', 'T', 'C'], [1/3, 1/3, 1/3]) nucl_choices['A'] = A nucl_choices['T'] = T nucl_choices['C'] = C nucl_choices['G'] = G nucl_choices_list.append(nucl_choices) return nucl_choices_list def dispatch_indels(read): """Return the x and y position of a insertion or deletion to be inserted in the indel matrix. The substitution matrix is a 2D array of size 301 * 9 The x axis (301) corresponds to the position in the read, while the y axis (9) represents the match or indel (see the dispatch dict in the function). Positions 0 is match or substitution, other positions in 'N1' are insertions, 'N2 are deletions' The size of x axis is 301 because we haven't calculated the read length yet Args: read (read): an aligned read object Yields: tuple: a tuple with the x, y position for dispatching the indel in the indel matrix """ logger = logging.getLogger(__name__) dispatch_indels = { 0: 0, 'A1': 1, 'T1': 2, 'C1': 3, 'G1': 4, 'A2': 5, 'T2': 6, 'C2': 7, 'G2': 8 } position = 0 for (cigar_type, cigar_length) in read.cigartuples: if cigar_type == 0: # match position += cigar_length continue elif cigar_type == 1: # insertion query_base = read.query_sequence[position] insertion = query_base.upper() + '1' try: indel = dispatch_indels[insertion] dispatch_tuple = (position, indel) position += cigar_length except KeyError as e: # we avoid ambiguous bases # logger.debug( # '%s not in dispatch: %s' % (insertion, e), exc_info=True) position += cigar_length continue elif cigar_type == 2: # deletion ref_base = read.query_alignment_sequence[position] deletion = ref_base.upper() + '2' try: indel = dispatch_indels[deletion] dispatch_tuple = (position, indel) position -= cigar_length except KeyError as e: # we avoid ambiguous bases # logger.debug( # '%s not in dispatch: %s' % (deletion, e), exc_info=True) position -= cigar_length continue else: logger.debug("CIGAR %s. Skipping read." % cigar_type) continue yield dispatch_tuple def indel_matrix_to_choices(indel_matrix, read_length): """Transform an indel matrix into probabilties of indels for at every position From the raw indel count at one position, returns a dictionary with probabilties of indel Args: indel_matrix (np.array): the substitution matrix is a 2D array of size read_length * 16. fhe x axis (read_length) corresponds to the position in the read, while the y axis (9) represents the match or indel. Positions 0 is match or substitution, other positions in 'N1' are insertions, 'N2 are deletions' read_length (int): read length Returns: tuple: tuple containing two lists of dictionaries representing the insertion or deletion probabilities for a collection of reads """ ins_choices = [] del_choices = [] for pos in range(read_length): insertions = { 'A': indel_matrix[pos][1] / indel_matrix[pos][0], 'T': indel_matrix[pos][2] / indel_matrix[pos][0], 'C': indel_matrix[pos][3] / indel_matrix[pos][0], 'G': indel_matrix[pos][4] / indel_matrix[pos][0] } deletions = { 'A': indel_matrix[pos][5] / indel_matrix[pos][0], 'T': indel_matrix[pos][6] / indel_matrix[pos][0], 'C': indel_matrix[pos][7] / indel_matrix[pos][0], 'G': indel_matrix[pos][8] / indel_matrix[pos][0] } ins_choices.append(insertions) del_choices.append(deletions) return (ins_choices, del_choices) InSilicoSeq-1.5.4/iss/profiles/000077500000000000000000000000001412502470100163205ustar00rootroot00000000000000InSilicoSeq-1.5.4/iss/profiles/HiSeq000066400000000000000000007734361412502470100173000ustar00rootroot00000000000000PKH[K2 LK\ model.npyPZ\nnj_TR_ wK)Ng$: (+!SPKH[KD.ZEXread_length.npyPZ\nni_TR_ wK)Ng$: (PKH[Kf,insert_size.npyw8p2* HQ"ie'BBFF]${d=mQFFs{{\yy\?q늚uJ E Gc;#[i^YSBֶm nZ`uEK;Tm`caaaa!A!^g^tRǒD^>+%Nq_ReHMըFΰ̓anHO^{) ?uo֗Q\޾ kS;&Fwjn&@pwxX>+:]:e^gh6ߥ`?:STBɟ\c,uhc$|nX4ɽ"VܥjpTD,4RvBJ, Yw$1*x7r_:2VF1~{P r'K`[/ɰl:0*ٷ G< -HImӹwi;`Z jEvpbCUhO[)04NEgXAeΩx G!7cV-o8 Iyl@񁦈2=N:U..|e{-&]Ln=*# ^тneX/^W9U6uKS0Wb{;¾J NEgŦJ$;(mFo*Y*] غg"U#75eÔ^AvmUT9ոLd5$(4Eϙ!ʍɊI.0eC]91=H"s̢M>1Kaw~ QXτ c|fݘWm-Jmkaw( )7ka˵@70![<ԉ4-|F!T }1ByXˉco7 @pҫk \ND>TrC% -}EwS x^WA !yI=M7.O+[VfP\6A>3lTQ.@+ȏߡ~4,7^UJ#\ By5C{Ny- \ݴ m_b`Åyo3ߎ> !7SN3a|,^3/̘8o)dnU}B=V'i&G}SGr{Q9Yj`lWW|y._odTlB 73c_~][w.~Vsfcӏn'rOG v0@~os5sw pV#^ҌsuZږ80\,AI8 ]fdtݐ%/oOd WxjP~\SGᠣ9 xgddG)~^15KM7?3.VDz["8Fa;Aԋdz<c4zT2~R=x3VUmA<<:R~ Ol}mȭ ;>!hJA{!DYz(O`Vzt6-iIi}MU7nxIo^k f뻎o4.Brp!Yxess[P?wLt?>NRl PPc$J#9. 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CX0ǴcwW'̺oaaN0 @{LQjbU cVemJ/n.U* V55-fzZRx]zaZ 777o:e3ކA7փn7Dpq ڄ07<ؒp#nmm8PWȕ-=A#jWrYm0 k`]ك*ZnH;c=` to{r7Г\w)N3ĝo69sgyD73W43[ &zkεz̵>3:f÷FM3ff6*WZsPKNqZK2 LK\ model.npyPKNqZKFXrread_length.npyPKNqZK}insert_size.npyPKNqZK2Opmean_count_forward.npyPKNqZKdyQpcmean_count_reverse.npyPKNqZK MOhquality_hist_forward.npyPKNqZKȍ7quality_hist_reverse.npyPKNqZKrI72ksubst_choices_forward.npyPKNqZK?PDsubst_choices_reverse.npyPKNqZK#DB+`ins_forward.npyPKNqZKK c+)0ins_reverse.npyPKNqZK3W+8҅\del_forward.npyPKNqZKCG8*$del_reverse.npyPK GInSilicoSeq-1.5.4/iss/test/000077500000000000000000000000001412502470100154545ustar00rootroot00000000000000InSilicoSeq-1.5.4/iss/test/test_abundance.py000066400000000000000000000070001412502470100210020ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import util from iss import abundance from nose.tools import raises, assert_almost_equal, with_setup import numpy as np def setup_function(): output_file_prefix = 'data/.test' def teardown_function(): util.cleanup(['data/test_abundance.txt']) def test_parsing(): abundance_dic = abundance.parse_abundance_file('data/abundance.txt') assert abundance_dic == { 'genome_ATCG': 0.1, 'genome_TA': 0.1, 'genome_A': 0.2, 'genome_GC': 0.4, 'genome_T': 0.2 } @raises(SystemExit) def test_parsing_empty(): abundance_dic = abundance.parse_abundance_file('data/empty_file') @raises(SystemExit) def test_parsing_no_exists(): abundance_dic = abundance.parse_abundance_file('data/does_not_exist') @raises(SystemExit) def test_parsing_bad_abundance(): abundance_dic = abundance.parse_abundance_file('data/bad_abundance.txt') def test_cov_calc(): coverage_ecoli = abundance.to_coverage( 10000000, 0.08, 150, 4639221 ) assert round(coverage_ecoli, 3) == 25.866 def test_distributions(): np.random.seed(42) f = open('data/genomes.fasta', 'r') with f: # count the number of records record_list = util.count_records(f) uniform_dic = abundance.uniform(record_list) halfnormal_dic = abundance.halfnormal(record_list) exponential_dic = abundance.exponential(record_list) lognormal_dic = abundance.lognormal(record_list) np.random.seed(42) # reset the seed to get 0s in zero_inflated_lognormal zero_inflated_lognormal_dic = abundance.zero_inflated_lognormal( record_list) assert list(uniform_dic.values()) == [0.2] * 5 assert round(halfnormal_dic['genome_A'], 2) == 0.16 assert round(exponential_dic['genome_A'], 2) == 0.01 assert round(lognormal_dic['genome_T'], 2) == 0.19 assert zero_inflated_lognormal_dic['genome_T'] == 0.0 assert round(zero_inflated_lognormal_dic['genome_A'], 2) == 0.44 @with_setup(setup_function, teardown_function) def test_abunance_draft(): abundance_dic = {'genome_A': 0.15511887441170918, 'genome_T': 0.08220476760848751, 'genome_GC': 0.18039811160555874, 'genome_ATCG': 0.4329003045949206, 'genome_TA': 0.07468835777633397, 'contig_1': 0.02776920430880394, 'contig_2': 0.011490705231229217, 'contig_3': 0.03542967446295675} np.random.seed(42) f = open('data/genomes.fasta', 'r') with f: # count the number of records complete_genomes = util.count_records(f) draft_genomes = ['data/draft.fasta'] ab = abundance.draft( complete_genomes, draft_genomes, abundance.lognormal, 'data/test') for tv, v in zip(abundance_dic.values(), ab.values()): assert round(tv) == round(v) # assert_almost_equal(ab, abundance_dic) def test_coverage_scaling(): abundance_dic = abundance.parse_abundance_file('data/abundance.txt') coverage_dic = abundance.coverage_scaling(10000, abundance_dic, 'data/genomes.fasta', 25) assert coverage_dic == {'genome_A': 136.6120218579235, 'genome_T': 136.6120218579235, 'genome_GC': 273.224043715847, 'genome_ATCG': 68.30601092896175, 'genome_TA': 68.30601092896175 } InSilicoSeq-1.5.4/iss/test/test_bam.py000066400000000000000000000006631412502470100176310ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss import bam from nose.tools import raises import os import sys @raises(SystemExit) def test_read_fail(): bam_file = 'data/empty_file' bam_reader = bam.read_bam(bam_file) for read in bam_reader: print(read) def test_to_model(): bam_file = 'data/ecoli.bam' output = 'data/test_bam' bam.to_model(bam_file, output) os.remove(output + '.npz') InSilicoSeq-1.5.4/iss/test/test_download.py000066400000000000000000000013601412502470100206740ustar00rootroot00000000000000#!/usr/bin/env python2 # -*- coding: utf-8 -*- from Bio.Seq import Seq from Bio.SeqRecord import SeqRecord from iss import download from iss.util import cleanup from nose.tools import with_setup def setup_function(): output_file_prefix = 'data/.test' def teardown_function(): cleanup(['data/test_download.fasta']) @with_setup(setup_function, teardown_function) def download_to_fasta(): ftp_url = 'ftp://ftp.ncbi.nlm.nih.gov/genomes/all/GCF/000/737/615/GCF_000737615.1_ASM73761v1/GCF_000737615.1_ASM73761v1_genomic.fna.gz' download.assembly_to_fasta(ftp_url, 'data/test_download.fasta') @with_setup(setup_function, teardown_function) def test_ncbi(): genome_list = download.ncbi('bacteria', 2, 'data/test_download.fasta') InSilicoSeq-1.5.4/iss/test/test_error_model.py000066400000000000000000000045641412502470100214070ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss.error_models import ErrorModel, basic, kde, perfect from Bio.Seq import Seq from Bio.SeqRecord import SeqRecord from nose.tools import raises import random import numpy as np def test_perfect_phred(): err_mod = perfect.PerfectErrorModel() distribution = err_mod.gen_phred_scores(20, 'forward')[:10] assert distribution == [40, 40, 40, 40, 40, 40, 40, 40, 40, 40] def test_basic_phred(): np.random.seed(42) err_mod = basic.BasicErrorModel() distribution = err_mod.gen_phred_scores(20, 'forward')[:10] assert distribution == [23, 19, 25, 40, 19, 19, 40, 26, 18, 23] def test_kde_phred(): np.random.seed(42) err_mod = kde.KDErrorModel('data/ecoli.npz') distribution = err_mod.gen_phred_scores(err_mod.quality_reverse, 'reverse')[10:] assert distribution == [40, 40, 40, 40, 40, 40, 40, 40, 10, 10] def test_introduce_errors(): np.random.seed(42) err_mod = basic.BasicErrorModel() read = SeqRecord( Seq(str('AATGC' * 25)), id='read_1', description='test read' ) read = err_mod.introduce_error_scores(read, 'forward') qualities = read.letter_annotations["phred_quality"][:10] assert qualities == [40, 26, 40, 40, 25, 25, 40, 40, 22, 40] def test_mut_sequence(): random.seed(42) np.random.seed(42) err_mod = basic.BasicErrorModel() read = SeqRecord( Seq(str('AAAAA' * 25)), id='read_1', description='test read' ) read.letter_annotations["phred_quality"] = [5] * 125 read.seq = err_mod.mut_sequence(read, 'forward') assert str(read.seq[:10]) == 'AAAACAGAAA' def test_introduce_indels(): random.seed(42) np.random.seed(42) err_mod = basic.BasicErrorModel() err_mod.ins_for[1]['G'] = 1.0 err_mod.del_for[0]['A'] = 1.0 bounds = (5, 130) read = SeqRecord( Seq(str('ATATA' * 25)), id='read_1', description='test read' ) ref_genome = SeqRecord( Seq(str('ATATA' * 100)), id='ref_genome', description='test reference' ) read.seq = err_mod.introduce_indels( read, 'forward', ref_genome, bounds) assert len(read.seq) == 125 assert read.seq[:10] == 'ATGATAATAT' @raises(SystemExit) def test_bad_err_mod(): err_mod = kde.KDErrorModel('data/empty_file') InSilicoSeq-1.5.4/iss/test/test_generator.py000066400000000000000000000061621412502470100210600ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from __future__ import unicode_literals from iss import generator from iss.util import cleanup from iss.error_models import ErrorModel, basic, kde from Bio.Seq import Seq from Bio.SeqRecord import SeqRecord from nose.tools import with_setup, raises import os import sys import random import numpy as np # due to inconsistent seeding between python 2 and 3, some of the following # tests are disabled with python2 def setup_function(): output_file_prefix = 'data/.test' def teardown_function(): cleanup(['data/.test.iss.tmp.my_genome.0_R1.fastq', 'data/.test.iss.tmp.my_genome.0_R2.fastq']) @raises(SystemExit) def test_cleanup_fail(): cleanup('data/does_not_exist') @with_setup(setup_function, teardown_function) def test_simulate_and_save(): err_mod = basic.BasicErrorModel() ref_genome = SeqRecord( Seq(str('AAAAACCCCC' * 100)), id='my_genome', description='test genome' ) generator.reads(ref_genome, err_mod, 1000, 0, 'data/.test', 0, True) @with_setup(setup_function, teardown_function) def test_simulate_and_save_short(): err_mod = basic.BasicErrorModel() ref_genome = SeqRecord( Seq(str('AACCC' * 100)), id='my_genome', description='test genome' ) generator.reads(ref_genome, err_mod, 1000, 0, 'data/.test', 0, True) @raises(AssertionError) def test_small_input(): err_mod = kde.KDErrorModel('data/ecoli.npz') ref_genome = SeqRecord( Seq(str('AAAAACCCCC')), id='my_genome', description='test genome' ) generator.simulate_read(ref_genome, err_mod, 1, 0) def test_basic(): if sys.version_info > (3,): random.seed(42) np.random.seed(42) err_mod = basic.BasicErrorModel() ref_genome = SeqRecord( Seq(str('AAAAACCCCC' * 100)), id='my_genome', description='test genome' ) read_tuple = generator.simulate_read(ref_genome, err_mod, 1, 0) big_read = ''.join(str(read_tuple[0].seq) + str(read_tuple[1].seq)) assert big_read[-15:] == 'TTTTGGGGGTTTTTG' def test_kde(): if sys.version_info > (3,): random.seed(42) np.random.seed(42) err_mod = kde.KDErrorModel('data/ecoli.npz') ref_genome = SeqRecord( Seq(str('CGTTTCAACC' * 400)), id='my_genome', description='test genome' ) read_tuple = generator.simulate_read(ref_genome, err_mod, 1, 0) big_read = ''.join(str(read_tuple[0].seq) + str(read_tuple[1].seq)) assert big_read[:15] == 'CCGTTTCAACCCGTT' def test_kde_short(): if sys.version_info > (3,): random.seed(42) np.random.seed(42) err_mod = kde.KDErrorModel('data/ecoli.npz') ref_genome = SeqRecord( Seq(str('AAACC' * 100)), id='my_genome', description='test genome' ) read_tuple = generator.simulate_read(ref_genome, err_mod, 1, 0) big_read = ''.join(str(read_tuple[0].seq) + str(read_tuple[1].seq)) assert big_read == 'ACCAAACCAAACCAAACCAAGGTTTGGTTTGGTTTGGTGT' InSilicoSeq-1.5.4/iss/test/test_modeller.py000066400000000000000000000041701412502470100206720ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from builtins import next from iss import bam from iss import modeller from nose.tools import assert_almost_equals import sys import numpy as np def test_kde_qualities(): quality_distribution = [ [40, 30], [40, 30], [20, 20], [40, 10], [10, 10]] cdf_list = modeller.raw_qualities_to_histogram(quality_distribution) assert_almost_equals(cdf_list[0][-2], 0.500002794) assert cdf_list[-1][0] == 0.0 assert cdf_list[-1][-1] == 1 assert len(cdf_list) == 5 def test_substitutions(): subst_matrix = np.zeros([20, 16]) bam_file = 'data/substitutions_test.bam' bam_reader = bam.read_bam(bam_file) if sys.version_info > (3,): for _ in range(2): bam_reader.__next__() read = bam_reader.__next__() # read_1_2 else: for _ in range(2): bam_reader.next() read = bam_reader.next() # read_1_2 alignment = read.get_aligned_pairs(matches_only=True, with_seq=True) read_has_indels = False for base in alignment: pos, subst, read_has_indels = modeller.dispatch_subst( base, read, read_has_indels) subst_matrix[pos, subst] += 1 choices = modeller.subst_matrix_to_choices(subst_matrix, 20) assert read_has_indels is False assert subst_matrix[0][1] == 1 assert choices[0]['A'] == (['T', 'C', 'G'], [1.0, 0.0, 0.0]) def test_indels(): indel_matrix = np.zeros([20, 9]) bam_file = 'data/substitutions_test.bam' bam_reader = bam.read_bam(bam_file) if sys.version_info > (3,): for _ in range(8): bam_reader.__next__() read = bam_reader.__next__() # read_4_1 else: for _ in range(8): bam_reader.next() read = bam_reader.next() # read_4_1 for pos, indel in modeller.dispatch_indels(read): indel_matrix[pos, indel] += 1 for position in range(20): indel_matrix[position][0] = 5 insertion, deletion = modeller.indel_matrix_to_choices( indel_matrix, 20) assert round(insertion[6]['T'], 2) == 0.2 assert indel_matrix[6][2] == 1 InSilicoSeq-1.5.4/iss/test/test_util.py000066400000000000000000000103051412502470100200410ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from nose.tools import raises, with_setup from iss import util from Bio import SeqIO import random def setup_function(): output_file_prefix = 'data/.test' def teardown_function(): util.cleanup(['data/test_concat.iss.tmp.genomes.fasta']) def teardown_compress(): util.cleanup(['data/ecoli.fasta.gz']) def test_phred_conversions(): assert util.phred_to_prob(40) == 0.9999 assert util.phred_to_prob(30) == 0.999 assert util.phred_to_prob(20) == 0.99 assert util.phred_to_prob(10) == 0.9 assert util.prob_to_phred(0.9999) == 40 assert util.prob_to_phred(0.999) == 30 assert util.prob_to_phred(0.99) == 20 assert util.prob_to_phred(0.9) == 10 def test_rev_comp(): lowercase_seq = 'attgctat' uppercase_seq = 'CCGATTAC' assert util.rev_comp(lowercase_seq) == 'atagcaat' assert util.rev_comp(uppercase_seq) == 'GTAATCGG' def test_count_records(): f = open('data/genomes.fasta', 'r') with f: # count the number of records record_list = util.count_records(f) assert record_list == [ 'genome_A', 'genome_T', 'genome_GC', 'genome_ATCG', 'genome_TA'] @raises(SystemExit) def test_count_records_empty(): util.count_records('data/empty_file') def test_split_list(): test_range = range(10) test_list = list(range(10)) # tests on range assert util.split_list(test_range) == [range(10)] assert util.split_list(test_range, n_parts=2) == [ range(0, 5), range(5, 10) ] assert util.split_list(test_range, n_parts=3) == [ range(0, 3), range(3, 6), range(6, 10) ] # tests on lists assert util.split_list(test_list) == [list(range(10))] assert util.split_list(test_list, n_parts=2) == [ list(range(0, 5)), list(range(5, 10))] assert util.split_list(test_list, n_parts=3) == [ list(range(0, 3)), list(range(3, 6)), list(range(6, 10)) ] def test_convert_n_reads(): simple_number = '10000' kilo_lower = '100k' kilo_upper = '100K' mega_lower = '20m' mega_upper = '20M' giga_lower = '1.5g' giga_upper = '0.5G' assert util.convert_n_reads(simple_number) == 10000 assert util.convert_n_reads(kilo_lower) == 100000 assert util.convert_n_reads(kilo_upper) == 100000 assert util.convert_n_reads(mega_lower) == 20000000 assert util.convert_n_reads(mega_upper) == 20000000 assert util.convert_n_reads(giga_lower) == 1500000000 assert util.convert_n_reads(giga_upper) == 500000000 @raises(SystemExit) def test_convert_n_reads_bad_not_a_number(): not_valid = 'rocket0' util.convert_n_reads(not_valid) @raises(SystemExit) def test_convert_n_reads_bad_suffix(): not_valid = '0.12Q' util.convert_n_reads(not_valid) @raises(SystemExit) def test_genome_file_exists(): my_important_file = 'data/ecoli.fasta' util.genome_file_exists(my_important_file) def test_reservoir(): samples = [] genome_file = 'data/genomes.fasta' with open(genome_file, 'r') as f: record_list = util.count_records(f) n = 2 with open(genome_file, 'r') as f: fasta_file = SeqIO.parse(f, 'fasta') for record in util.reservoir(fasta_file, record_list, n): samples.append(record.id) assert len(samples) == 2 @raises(SystemExit) def test_reservoir_invalid_input(): genome_file = 'data/ecoli.fasta' record_list = ['NC_002695.1'] n = 4 with open(genome_file, 'r') as f: fasta_file = SeqIO.parse(f, 'fasta') for record in util.reservoir(fasta_file, record_list, n): pass @with_setup(setup_function, teardown_function) def test_concatenate(): genome_files = ['data/ecoli.fasta'] * 2 util.concatenate(genome_files, 'data/test_concat.iss.tmp.genomes.fasta') with open('data/test_concat.iss.tmp.genomes.fasta', 'rb') as f: assert len(f.readlines()) == 40 @raises(SystemExit) def test_concatenate_read_only(): genome_files = ['data/ecoli.fasta'] * 2 util.concatenate(genome_files, 'data/read_only.fasta') @with_setup(setup_function, teardown_compress) def test_compress(): genome_file = 'data/ecoli.fasta' util.compress(genome_file, remove=False) InSilicoSeq-1.5.4/iss/util.py000066400000000000000000000165431412502470100160350ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from Bio import SeqIO import os import sys import gzip import pickle import random import logging import numpy as np from shutil import copyfileobj def phred_to_prob(q): """Convert a phred score (Sanger or modern Illumina) in probabilty Given a phred score q, return the probabilty p of the call being right Args: q (int): phred score Returns: float: probabilty of basecall being right """ p = 10 ** (-q / 10) return 1 - p def prob_to_phred(p): """Convert a probabilty into a phred score (Sanger or modern Illumina) Given a probabilty p of the basecall being right, return the phred score q Args: p (int): probabilty of basecall being right Returns: int: phred score """ q = int(round(-10 * np.log10(1 - p))) return q def rev_comp(s): """A simple reverse complement implementation working on strings Args: s (string): a DNA sequence (IUPAC, can be ambiguous) Returns: list: reverse complement of the input sequence """ bases = { "a": "t", "c": "g", "g": "c", "t": "a", "y": "r", "r": "y", "w": "w", "s": "s", "k": "m", "m": "k", "n": "n", "b": "v", "v": "b", "d": "h", "h": "d", "A": "T", "C": "G", "G": "C", "T": "A", "Y": "R", "R": "Y", "W": "W", "S": "S", "K": "M", "M": "K", "N": "N", "B": "V", "V": "B", "D": "H", "H": "D"} sequence = list(s) complement = "".join([bases[b] for b in sequence]) reverse_complement = complement[::-1] return reverse_complement def count_records(fasta_file): """Count the number of records in a fasta file and return a list of recods id Args: fasta_file (string): the path to a fasta file Returns: list: a list of record ids """ logger = logging.getLogger(__name__) record_list = [] for record in SeqIO.parse(fasta_file, "fasta"): record_list.append(record.id) try: assert len(record_list) != 0 except AssertionError as e: logger.error( 'Failed to find records in genome(s) file:%s' % fasta_file) sys.exit(1) else: return record_list def split_list(l, n_parts=1): """Split a list in a number of parts Args: l (list): a list n_parts (in): the number of parts to split the list in Returns: list: a list of n_parts lists """ length = len(l) return [l[i * length // n_parts: (i + 1) * length // n_parts] for i in range(n_parts)] def nplog(type, flag): logger = logging.getLogger(__name__) logger.debug("FloatingPointError (%s), with flag %s" % (type, flag)) def convert_n_reads(unit): """For strings representing a number of bases and ending with k, K, m, M, g, and G converts to a plain old number Args: n (str): a string representing a number ending with a suffix Returns: float: a number of reads """ logger = logging.getLogger(__name__) suffixes = {'k': 3, 'm': 6, 'g': 9} if unit[-1].isdigit(): try: unit_int = int(unit) except ValueError as e: logger.error('%s is not a valid number of reads' % unit) sys.exit(1) elif unit[-1].lower() in suffixes: number = unit[:-1] exponent = suffixes[unit[-1].lower()] unit_int = int(float(number) * 10**exponent) else: logger.error('%s is not a valid number of reads' % unit) sys.exit(1) return unit_int def genome_file_exists(filename): """Checks if the output file from the --ncbi option already exists Args: filename (str): a file name """ logger = logging.getLogger(__name__) try: assert os.path.exists(filename) is False except AssertionError as e: logger.error('%s already exists. Aborting.' % filename) logger.error('Maybe use another --output prefix') sys.exit(1) def reservoir(records, record_list, n=None): """yield a number of records from a fasta file using reservoir sampling Args: records (obj): fasta records from SeqIO.parse Yields: record (obj): a fasta record """ logger = logging.getLogger(__name__) if n is not None: try: total = len(record_list) assert n < total except AssertionError as e: logger.error( '-u should be strictly smaller than total number of records.') sys.exit(1) else: random.seed() x = 0 samples = sorted(random.sample(range(0, total - 1), n)) for sample in samples: while x < sample: x += 1 _ = records.__next__() record = records.__next__() x += 1 yield record else: for record in records: yield record def concatenate(file_list, output): """Concatenate files together Args: file_list (list): the list of input files (can be a generator) output (string): the output file name """ logger = logging.getLogger(__name__) logger.info('Stitching input files together') try: out_file = open(output, 'wb') except (IOError, OSError) as e: logger.error('Failed to open output file: %s' % e) sys.exit(1) with out_file: for file_name in file_list: if file_name is not None: with open(file_name, 'rb') as f: copyfileobj(f, out_file) def cleanup(file_list): """remove temporary files Args: file_list (list): a list of files to be removed """ logger = logging.getLogger(__name__) logger.info('Cleaning up') for temp_file in file_list: if temp_file is not None: try: os.remove(temp_file) except (IOError, OSError) as e: logger.error('Could not read temporary file: %s' % temp_file) logger.error('You may have to remove temporary files manually') sys.exit(1) def compress(filename, remove=True): """gzip a file Args: filename (string): name of file to be compressed """ logger = logging.getLogger(__name__) logger.info('Compressing %s' % filename) outfile = filename + '.gz' with open(filename, 'rb') as i, gzip.open(outfile, 'wb') as o: copyfileobj(i, o) if remove: cleanup([filename]) return outfile def dump(object, output): """dump an object, like pickle.dump. This function uses pickle.dumps to dump large objects Args: object (object): a python object """ MAX_BYTES = 2**31 - 1 pickled_object = pickle.dumps(object, protocol=pickle.HIGHEST_PROTOCOL) size = sys.getsizeof(pickled_object) with open(output, 'wb') as out_file: for i in range(0, size, MAX_BYTES): out_file.write(pickled_object[i:i + MAX_BYTES]) def load(filename): """load a pickle from disk This function uses pickle.loads to load large objects Args: filename (string): the path of the pickle to load """ MAX_BYTES = 2**31 - 1 size = os.path.getsize(filename) bytes = bytearray(0) with open(filename, 'rb') as f: for _ in range(0, size, MAX_BYTES): bytes += f.read(MAX_BYTES) object = pickle.loads(bytes) return object InSilicoSeq-1.5.4/iss/version.py000066400000000000000000000000261412502470100165320ustar00rootroot00000000000000__version__ = '1.5.4' InSilicoSeq-1.5.4/setup.py000066400000000000000000000016141412502470100154130ustar00rootroot00000000000000#!/usr/bin/env python # -*- coding: utf-8 -*- from iss.version import __version__ from setuptools import setup, find_packages url = 'https://github.com/HadrienG/InSilicoSeq' with open('README.md') as f: long_description = f.read() setup( name='InSilicoSeq', version=__version__, description='a sequencing simulator', long_description=long_description, long_description_content_type='text/markdown', url=url, download_url=url + '/tarball/' + __version__, author='Hadrien Gourlé', author_email='hadrien.gourle@slu.se', license='MIT', packages=find_packages(), tests_require=['nose'], install_requires=['numpy', 'scipy', 'biopython<=1.78', 'pysam>=0.15.1', 'future', 'joblib', 'requests'], include_package_data=True, entry_points={ 'console_scripts': ['iss = iss.app:main'], } )

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