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FastaHandler

A collection of Python scripts designed for the efficient management of various FASTA file formats.

FASTAhandler Logo

Brief Background

FastaHandler, created by Hyungtaek Jung and the team at the National Centre for Indigenous Genomics at The Australian National University, is a Python script suite for efficient FASTA file management. It boasts 19 work modules to ease input/output processes, covering various aspects of FASTA data analysis including post-processing and format conversion. Optimized for life science datasets, FastaHandler is a CLI application tested across different FASTA formats. Users should note that processing large datasets may require substantial computational resources on Linux, HPC or Cloud platforms.

Citation

Hyungtaek Jung et al. 2024: FastaHandler: An easy Python-based toolset for handling fasta files, PLoS Comp Biol Submitted.

Contents:

STABLE (version 1.0.1)

  • Release date: January 2024
  • FastaHandler is a standalone Python application equipped with 18 sub-modules for interactive FASTA file manipulation, available as open-source (see LICENSE).

INSTALLATION

  • Access the program via GitHub
  • Installation options include Bioconda or Python pip. For support, refer to Issues on GitHub.
  • FastaHandler requires no separate installation process.
  • Just clone this repository, and run
git clone https://github.com/OZTaekOppa/FASTAhandler/
python3 {path}/fastahandler.py

LICENSE

FastaHandler is available under the MIT license and incorporates various open-source software. For detailed information on the integrated Python packages, modules, and libraries, and their specific applications within FastaHandler, please refer to the manuscript

Tested Datasets

Please refer to the example dataset folder for sample data and usage demonstrations.

GETTING STARTED

FastaHandler is developed primarily in Python 3.9+ and Biopython and features 19 modules. It facilitates data input and output through a Command-Line Interface (CLI), ensuring smooth end-to-end file handling. To optimize the use of FastaHandler, users should prepare all necessary input files, such as FASTA and TXT formats, in advance.

FASTAhandler Workflow

General Usage

FastaHandler: Fasta File Manipulation Toolkit
version 1.0.1

Usage: python3 fastahandler.py <module> <parameters>

Modules:
Multi2Single    		| m2s   	Convert a multi-fasta (multiline) into a single-line fasta.
Gfa2Fasta	    		| g2a   	Convert a gfa (Graphical Fragment Assembly) into a single-line fasta.
RenameId        		| rid   	Rename prefix IDs and headers.
PrefixRename    		| prn   	Rename prefix IDs and headers with a user’s input.
PrefixSelectRename      	| psr   	Rename prefix IDs and headers with a user’s input (Only).
PrefixFindReplaceRename      	| pfr   	Replace and rename prefix IDs and headers with a user’s input (Only).
IdExtract       		| idx   	Extract matched IDs and their corresponding sequences.
IdExtractLocation       	| iel   	Extract matched IDs, locations and their corresponding sequences.
IdExtractLocationMultiple       | iem   	Extract matched IDs, locations and their corresponding sequences (Multiple).
ReverseComplement       	| rcp   	Make a reverse complement sequence.
FindCountDuplication    	| fcd   	Find and count the duplicated IDs and sequences.
RemoveDuplication       	| rvd   	Remove the duplicated IDs and sequences.
SubsetFasta     		| ssf   	Make a subset of data with a sequence length filter.
ExtractPattern  		| xpt   	Make a subset of data with find, filter and extract.
EachFastaStats  		| efs   	Generate each line fasta statistic for a multi-line fasta.
AllFastaStats   		| afs   	Generate a summary of multi-line fasta statistics.
MultipleFastaStats      	| mfs   	Generate a summary of multi-line fasta statistics (Multiple).
ConcatenateFasta        	| ccf   	Make a concatenated fasta file (Multiple).
TranslateSequence       	| tls   	Find the translated sequences as a protein and open reading frames (ORFs).

Use <module> --help for module usage.

multi2single (m2s)

  • To convert a multi-fasta (multiline) into a single-line fasta.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A multiline fasta file.
    • Output: A single-line fasta.
    • Example file: mltseq2sl in the "example_data" folder.

Example usage

python3 multi2single.py --input-seq test_dna.fasta --out test_output_sl.fasta --t 1 --mem 2
  • Parameter explanation
    1. python 3: Call python 3
    2. multi2single.py: Call multi2single module
    3. python3 multi2singleline.py --help: Check help menu
      • --input-seq: Indicate an input multi-fasta (multiline) fasta file and its path
      • --out: Indicate an output single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only only with Gb size)

gfa2fa (g2a)

  • To convert a gfa (Graphical Fragment Assembly) into a single-line fasta.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A gfa file.
    • Output: A single-line fasta.
    • Example file: gfa2fa in the "example_data" folder.

Example usage

python3 gfa2fa.py --input-gfa test_dna.gfa --out test_output_sl.fasta --t 1 --mem 2
  • Parameter explanation
    1. python 3: Call python 3
    2. gfa2fa.py: Call gfa2fa module
    3. python3 gfa2fa.py --help: Check help menu
      • --input-gfa: Indicate an input gfa file and its path
      • --out: Indicate an output single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only only with Gb size)

renameid (rid)

  • To rename prefix IDs and headers from a single-line fasta.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file.
    • Output: A single-line fasta with a new prefix ID name.
    • Example file: renameid_seq.fa in the "example_data" folder.

Example usage

python3 renameid.py --input-seq test_dna.fasta --new-name FunNGS --out output_reN.fasta --t 1 --mem 2
  • Use the multi2single module first if your input FASTA file isn't in single-line format.
  • Parameter explanation
    1. python 3: Call python 3
    2. renameid.py: Call renameid module
    3. python3 renameid.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --new-name: Indicate a new prefix ID/header name (accept both integer and strings but no space)
      • --out: Indicate an output renamed single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

prfxrename (prn)

  • To rename prefix IDs and headers from a single-line fasta with a user's input text file.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file and a tap-separated text file (e.g. old_IDs new_IDs)
    • Output: A single-line fasta with a new prefix ID name based on the user's input text file. Unmatched IDs will be also produced with its original IDs.
    • Example file: header_id_only.txt in the "example_data" folder.

Example usage

python3 prfxrename.py --input-seq test_dna.fasta --input-id new_ids.txt --out output_reN.fasta --t 1 --mem 2
  • Although the script can process multiline FASTA files, if you're unsure about your input FASTA file format, it's recommended to first convert it to single-line format using the multi2single module.
  • The script is ideal for renaming specific parts of IDs/headers in FASTA files, such as those from genome assemblies acquired from public databases like NCBI and EBI, based on user input.
  • Parameter explanation
    1. python 3: Call python 3
    2. prfxrename.py: Call prfxrename module
    3. python3 renameid.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --input-id: Indicate a new tap-separated prefix ID/header name file (accept both integer and strings but no space)
      • --out: Indicate an output renamed single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

prfxselrename (psr)

  • To rename prefix IDs and headers from a single-line fasta with a user's input text file.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file and a tap-separated text file (e.g. old_IDs new_IDs)
    • Output: A single-line fasta with a new prefix ID name based on the user's input text file. Unmatched IDs will be discarded.
    • Example file: header_id_only.txt in the "example_data" folder.

Example usage

python3 prfxselrename.py --input-seq test_dna.fasta --input-id new_ids.txt --out output_reN.fasta --t 1 --mem 2
  • Although the script can process multiline FASTA files, if you're unsure about your input FASTA file format, it's recommended to first convert it to single-line format using the multi2single module.
  • The script is effective for selectively renaming IDs/headers in FASTA files (with a user's specific input), such as genome assemblies from databases, where you want to exclude and not rename certain elements like scaffolds.
  • Parameter explanation
    1. python 3: Call python 3
    2. prfxselrename.py: Call prfxselrename module
    3. python3 prfxselrename.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --input-id: Indicate a new tap-separated prefix ID/header name file (accept both integer and strings but no space)
      • --out: Indicate an output renamed single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

prfxfindreplace (pfr)

  • To rename prefix IDs and headers from a single-line fasta with a user's input text file.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file and a find and replace pattern (e.g. old_IDs new_IDs)
    • Output: A single-line fasta with a new prefix ID name based on the user's input text file. Worked as "sed" command.
    • Example file: header_id_only.txt in the "example_data" folder.

Example usage

python3 prfxfindreplace.py --input-seq --find-ptrn hifiasm --replace Assembly --out output_reN.fasta --t 1 --mem 2
  • Although the script can process multiline FASTA files, if you're unsure about your input FASTA file format, it's recommended to first convert it to single-line format using the multi2single module.
  • The script is effective for selectively renaming IDs/headers in FASTA files (with a user's specific input), such as genome assemblies from databases, where you want to exclude and not rename certain elements like scaffolds.
  • Parameter explanation
    1. python 3: Call python 3
    2. prfxfindreplace.py: Call prfxfindreplace module
    3. python3 prfxfindreplace.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --find-ptrn: Indicate a pattern in prefix ID/header name file (accept both integer and strings but no space)
      • --replace: Indicate a new prefix ID/header name for replacement
      • --out: Indicate an output renamed single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

idextract (idx)

  • To extract matched IDs and their corresponding sequences.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file and a list of IDs.
    • Output: A single-line fasta with extracted IDs and their corresponding sequences.
    • Example file: header_id.txt and idextract.fa in the "example_data" folder.

Example usage

python3 idextract.py --input-seq test_dna.fasta --input-hdr header_id.txt --out output_extracted.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. idextract.py: Call idextract module
    3. python3 idextract.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --input-header: Indicate an input ID and header (without ">") text file and its path
      • --out: Indicate an output ID matched and extracted single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

idextloct (iel)

  • To extract matched IDs, locations and their corresponding sequences (focused on a single ID)
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file and a list of IDs.
    • Output: A single-line fasta with extracted IDs and their corresponding sequences.
    • Example file: header_id.txt and idextloct.fa in the "example_data" folder.

Example usage

python3 idextloct.py --input-seq test_dna.fasta --header-id test3_3%week --start 2 --end 10 --out output_test.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. idextloct.py: Call idextloct module
    3. python3 idextloct.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --header-id: Indicate an input ID and header (without ">") name and pattern
      • --start: Indicate a start position to extract (please use -1 value due to the python index)
      • --end: Indicate en end position to extract (please use -1 value due to the python index)
      • --out: Indicate an output ID matched and extracted single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

idextloctmlt (iem)

  • To extract matched IDs, locations and their corresponding sequences (focused on multiple IDs)
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file and a list of IDs.
    • Output: A single-line fasta with extracted IDs and their corresponding sequences.
    • Example file: hdrmulti_id.txt, idextract.fa and idextloct.fa in the "example_data" folder.

Example usage

python3 idextloctmlt.py --input-seq test_dna.fasta --input-extract input_extract.txt --out output_extest.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. idextloctmlt.py: Call idextloctmlt module
    3. python3 idextloctmlt.py --help: Check help menu --input-seq: Indicate an input single-line fasta file and its path --input-extract: Indicate an input ID and header (without ">") text file (a tap-separated) and its path including start and end positions (please use -1 value due to the python index) --out: Indicate an output ID matched and extracted single-line fasta file and its path --t: Specify thread numbers (integer only) --mem: Specify memory numbers (integer only with Gb size)

revcomplt (rcp)

  • To make a reverse complement sequence
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file.
    • Output: A reverse complement single-line fasta.
    • Example file: revscomplt.fa in the "example_data" folder.

Example usage

python3 revcomplt.py --input-seq test_dna.fasta --out output_revctest.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. revcomplt.py: Call revcomplt module
    3. python3 revcomplt.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path
      • --out: Indicate a reverse complement converted output single-line fasta file and its path
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

findcntdupl (fcd)

  • To find and count the duplicated IDs and sequences from a multi-fasta file
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file.
    • Output: A text with a duplication number.
    • Example file: dupids.fa in the "example_data" folder.

Example usage

python3 findcntdupl.py --input-seq test_dna2.fasta --out output_files.txt --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. findcntdupl.py: Call findcntdupl module
    3. python3 findcntdupl.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path (accept reverse complement sequences)
      • --out: Indicate an output text file after finding and counting the duplicated IDs and sequences (only for both matched IDs and their corresponding sequences)
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

removedupl (rvp)

  • To remove the duplicated IDs and sequences from a multi-fasta file
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file.
    • Output: A single-line fasta after removing updated IDs and sequences.
    • Example file: rmvdup.fa in the "example_data" folder.

Example usage

python3 removedupl.py --input-seq test_dna2.fasta --outfasta output_testdupl.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. removedupl.py: Call removedupl module
    3. python3 removedupl.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path (accept reverse complement sequences)
      • --outfasta: Indicate an output fasta file after removing the duplicated IDs and sequences (only for both matched IDs and their corresponding sequences)
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

subsetfa (ssf)

  • To make a subset of data with a sequence length filter option
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A fasta file.
    • Output: A subseted single-line fasta.
    • Example file: subsetfa.fa in the "example_data" folder.

Example usage

python3 subsetfa.py --input-seq test_mRNA1.fasta --filter 50 --out output_subset.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. subsetfa.py: Call subsetfa module
    3. python3 subsetfa.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path (accept reverse complement sequences)
      • --filter: Indicate a length size to filter out
      • --out: Indicate an output fasta file after filtering the length size (integer only)
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

extractptrn (xpt)

  • To make a subset of data with find, filter and extract options

Example usage

python3 extractptrn.py --input-seq test_dna1.fasta --input-pattern seq_pattern.txt --input-length 45 --out output_pattern.fasta --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. extractptrn.py: Call extractptrn module
    3. python3 extractptrn.py --help: Check help menu
      • --input-seq: Indicate an input single-line fasta file and its path (accept reverse complement sequences)
      • --input-pattern: Indicate an input text file and its path to find and filter a specific sequence pattern (accept reverse complement sequences)
      • --input-length: Indicate a length size to filter out
      • --out: Indicate an output fasta file after filtering the length size (integer only)
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

eachfastats (efs)

  • To generate each line fasta statistic for a multi-line fasta
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A multi-line fasta file.
    • Output: A summary of single-line fasta with its corresponding sequence length.
    • Example file: all_stat_asm.fa and stat_asm_mlt_unlm.fa in the "example_data" folder.

Example usage

python3 eachfastats.py --input-seq test_dna.fasta --out output_dna.txt --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Parameter explanation
    1. python 3: Call python 3
    2. eachfastats.py: Call eachfastats module
    3. python3 eachfastats.py --help: Check help menu
      • --input-seq: Indicate an input multi-line fasta file and its path
      • --out: Indicate an output text file with the sequence length
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

allfastats (afs)

  • To generate a summary of multi-line fasta statistics.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: A multi-line fasta file.
    • Output: A summary of multi-line fasta with its diverse statistics (e.g. sequence length, GC content, N50, and more).
    • Example file: all_stat_asm.fa and stat_asm_mlt_unlm.fa in the "example_data" folder.

Example usage

python3 allfastats.py --input-seq test_dna.fasta --out output_dna.txt --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Both eachfastats and allfastasts modules use the same multi-line FASTA input. Eachfastats focuses on individual FASTA lines and their sequence lengths, while allfastasts provides a summary of assembly statistics, such as for genomes or transcriptomes.
  • Parameter explanation
    1. python 3: Call python 3
    2. allfastats.py: Call allfastats module
    3. python3 allfastats.py --help: Check help menu
      • --input-seq: Indicate an input multi-line fasta file and its path
      • --out: Indicate an output text file with the summary of statistics
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

asmstatsunlm (mfs)

  • To generate a summary of multi-line fasta statistics for unlimited fasta files. An extended version of the allfastats for multiple fasta files.
    • Requirement: The script of Python/bash requires a Python library.
    • Input: Unlimited multi-line fasta files.
    • Output: A summary of multi-line fasta with its diverse statistics (e.g. sequence length, GC content, N50, and more).
    • Example file: all_stat_asm.fa and stat_asm_mlt_unlm.fa in the "example_data" folder.

Example usage

python3 asmstatsunlm.py --input-seqs test_dna1.fasta test_dna2.fasta test_dna3.fasta test_dna4.fasta test_dna5.fasta --out output_dna.txt --t 1 --mem 2
  • If your input FASTA file is in multi-line format, the script will automatically convert it to single-line format for processing (embedded pipeline).
  • Both allfastasts and asmstatsunlm modules take the same multi-line FASTA files. Allfastats generates a summary of assembly statistics from a single FASTA file, while asmstatsunlm does so for multiple FASTA files, useful for genome or transcriptome analysis.
  • Parameter explanation
    1. python 3: Call python 3
    2. asmstatsunlm.py: Call asmstatsunlm module
    3. python3 asmstatsunlm.py --help: Check help menu
      • --input-seqs: Indicate an input multi-line fasta file and its path (Separated by a space for each new fasta file)
      • --out: Indicate an output text file with the summary of statistics
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

concatenate (ccf)

  • To make a concatenated fasta file for unlimited fasta files.

Example usage

python concatenate.py --input-seqs test_dna1.fasta test_dna2.fasta test_dna3.fasta test_dna4.fasta test_dna5.fasta --out output_concat.fasta --t 1 --mem 2
  • For optimal use of this module, ensure that all input FASTA files have matching prefix IDs and headers and are formatted as single-line FASTA before concatenation.
  • Before using the concatenate module, ensure your FASTA files are in single-line format by using the renameid and multi2singleline modules, even though the pipeline automatically converts multi-line FASTA files.
  • Parameter explanation
    1. python 3: Call python 3
    2. concatenate.py: Call concatenate module
    3. python3 concatenate.py --help: Check help menu
      • --input-seqs: Indicate input multiple single-line fasta files and their path (Separated by a space for each new fasta file)
      • --out: Indicate a concatenated output fasta file and its path
      • --t: Specify thread numbers (intergr only)
      • --mem: Specify memory numbers (integer only with Gb size)

translated (tls)

  • To find the translated sequences as a protein including the open reading frames (ORFs)
    • Requirement: The script of Python/bash requires a Python library.
    • Input: Multline fasta files.
    • Output: A single-line fasta with translation and selected ORFs (nucleotide and protein sequences).
    • Example file: trnsldna.fa in the "example_data" folder.

Example usage

python translatedna.py --input-seq test_dna.fasta --out output/folder --t 1 --mem 2
  • For optimal use of this module, ensure that all input FASTA files have matching prefix IDs and headers and are formatted as single-line FASTA before concatenation.
  • Before using the translatedna module, ensure your FASTA files are in single-line format by using the renameid and multi2singleline modules, even though the pipeline automatically converts multi-line FASTA files.
  • Parameter explanation
    1. python 3: Call python 3
    2. translatedna.py: Call translatedna module
    3. python3 translatedna.py --help: Check help menu
      • --input-seq: Indicate input single-line fasta files and their path
      • --out: Indicate a translated output fasta file and its path (a total of four fasta files for both nucleotide and protein sequences)
      • --t: Specify thread numbers (integer only)
      • --mem: Specify memory numbers (integer only with Gb size)

FAQ

We encourage users to use the Issues.

WIKI PAGE

Please see the GitHub page.

AUTHORS

Hyungtaek Jung and the National Centre for Indigenous Genomics.

COPYRIGHT

The full FastaHandler is distributed under the MIT license.

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Python scripts designed for efficiently managing various types of FASTA file formats

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