#Dependencies import os, sys, re import shutil import argparse def get_args(): parser = argparse.ArgumentParser( prog = 'PhyloToL v6.0 Part 1 for Transcriptomes', description = "Updated September 29th, 2023 by Auden Cote-L'Heureux. Link to GitHub: https://github.com/AudenCote/PhyloToL_v6.0" ) parser.add_argument('-s', '--script', default = -1, type = int, choices = { 1, 2, 3, 4, 5, 6, 7 }, help = 'Script to run if you are only running one script') parser.add_argument('-n', '--conspecific_names', type = str, help = 'A .txt or .tsv file with two tab-separated columns; the first should have 10 digit codes, the second species or other identifying names. This is used to determine which sequences to remove (only between "species") in cross-plate contamination assessment') parser.add_argument('-1', '--first_script', default = -1, type = int, choices = { 1, 2, 3, 4, 5, 6 }, help = 'First script to run') parser.add_argument('-2', '--last_script', default = -1, type = int, choices = { 2, 3, 4, 5, 6, 7 }, help = 'First script to run') parser.add_argument('-a', '--assembled_transcripts', type = str, help = 'Path to a folder of assembled transcripts, assembled by rnaSPAdes. Each assembled transcript file name should start with a unique 10 digit code, and end in "_assembledTranscripts.fasta", E.g. Op_me_hsap_assembledTranscripts.fasta') parser.add_argument('-o', '--output', default = '../', type = str, help = 'An "Output" folder will be created at this directory to contain all output files. By default this folder will be created at the parent directory of the Scripts folder') parser.add_argument('-x', '--xplate_contam', action = 'store_true', help = 'Run cross-plate contamination removal (includes all files)') parser.add_argument('-g', '--genetic_code', type = str, help = 'If all of your taxa use the same genetic code, you may enter it here (to be used in script 5). Alternatively, if you need to use a variety of genetic codes but know which codes to use, you may fill give here the path to a .txt or .tsv with two tab-separated columns, the first with the ten-digit codes and the second column with the corresponding genetics codes. Otherwise, stop at script 4 and fill in "gcode_output.tsv" before running script 5') parser.add_argument('-min', '--minlen', type = int, default = 200, help = 'Minimum transcript length') parser.add_argument('-max', '--maxlen', type = int, default = 12000, help = 'Maximum transcript length') parser.add_argument('-d', '--databases', type = str, default = '../Databases', help = 'Path to databases folder') return parser.parse_args() #running the first script on all the bare files def script_one(args, ten_digit_codes): for file in os.listdir(args.assembled_transcripts): if file[10:] == '_assembledTranscripts.fasta' and file[:10] in ten_digit_codes: os.system('python 1a_TranscriptLengthFilter.py --input_file ' + args.assembled_transcripts + '/' + file + ' --output_file ' + args.output + '/Output/' + file[:10] + ' --minLen ' + str(args.minlen) + ' --maxLen ' + str(args.maxlen) + ' --spades') #SPADES ARGUMENT?? if args.xplate_contam: os.system('python 1b_CrossPlateContamination.py ' + args.output + '/Output/XlaneBleeding ' + str(args.minlen) + ' ' + args.conspecific_names) def script_two(args): for folder in os.listdir(args.output + '/Output/'): if os.path.isfile(args.output + '/Output/' + folder + '/SizeFiltered/' + folder + '.' + str(args.minlen) + 'bp.fasta'): os.system('python 2a_Identify_rRNA.py --input_file ' + args.output + '/Output/' + folder + '/SizeFiltered/' + folder + '.' + str(args.minlen) + 'bp.fasta --databases ' + args.databases) fasta_withBact = args.output + '/Output/' + folder + '/' + folder + '_NorRNAseqs.fasta' os.system('python 2b_Identify_Proks.py --input_file ' + fasta_withBact + ' --databases ' + args.databases) #NEED TO SORT OUT FILE NAMES ETC. BELOW HERE #running the third script def script_three(args): for folder in os.listdir(args.output + '/Output'): if os.path.isfile(args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.fasta'): os.system('python 3_AssignOGs.py --input_file ' + args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.fasta --evalue 1e-15 --databases ' + args.databases) #running the fourth script def script_four(args): for folder in os.listdir(args.output + '/Output'): if os.path.isfile(args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.Renamed.fasta'): os.system('python 4_InFrameStopCodonEstimator.py --input_file ' + args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.Renamed.fasta --databases ' + args.databases) #putting all of the gcode summaries produced by fourth script into a spreadsheet gcode_info = [] for folder in os.listdir(args.output + '/Output'): if os.path.isfile(args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.Renamed_StopCodonStats.tsv'): with open(args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.Renamed_StopCodonStats.tsv') as f: for line in f: line_sep = line.split('\t') if line_sep[0] == 'Summary': gcode_info.append([folder, line_sep[6], line_sep[7], line_sep[8][:-1]]) valid_codes = ['bleph','blepharisma','chilo','chilodonella','condy', 'condylostoma','none','eup','euplotes','peritrich','vorticella','ciliate','universal','taa','tag','tga','mesodinium'] stop = False gcode_file = { } if args.genetic_code.endswith('.txt') or args.genetic_code.endswith('.tsv'): if os.path.isfile(args.genetic_code): for line in open(args.genetic_code): try: if line.split('\t')[1].strip().lower() in valid_codes: gcode_file.update({ line.split('\t')[0] : line.split('\t')[1].strip() }) else: print('Genetic code ERROR -- ' + line.split('\t')[1].strip() + ' is not a valid genetic code. Please fill out the "gcode_output.tsv" file and continue with script 5.') except IndexError: print('\nGenetic code ERROR -- it looks like you tried to enter a .txt/.tsv file, but it is improperly formatted. Stopping after script 4; you may fill out the file gcode_output.tsv and continue with script 5.\n') stop = True else: print('\nGenetic code ERROR -- it looks like you tried to enter a .txt/.tsv file, but it could not be found. Stopping after script 4; you may fill out the file gcode_output.tsv and continue with script 5.\n') stop = True with open(args.output + '/Output/gcode_output.tsv', 'w') as g: g.writelines('10 Digit Code\tIn-frame TAG Density\tIn-frame TGA Density\tIn-frame TAA Density\tGenetic Code\n') for row in gcode_info: if args.genetic_code == None: g.writelines(row[0] + '\t' + row[1] + '\t' + row[2] + '\t' + row[3] + '\t\n') elif args.genetic_code.lower() in valid_codes: g.writelines(row[0] + '\t' + row[1] + '\t' + row[2] + '\t' + row[3] + '\t' + args.genetic_code + '\n') elif args.genetic_code.endswith('.txt') or args.genetic_code.endswith('.tsv'): try: g.writelines(row[0] + '\t' + row[1] + '\t' + row[2] + '\t' + row[3]+ '\t' + gcode_file[row[0]] + '\n') except KeyError: g.writelines(row[0] + '\t' + row[1] + '\t' + row[2] + '\t' + row[3]+ '\t' + 'Universal' + '\n') print('\nDefaulting to Universal genetic code for taxon ' + row[0] + '\n') #print('\nGenetic code ERROR -- it looks like you tried to enter a .txt/.tsv file, but a taxon is missing. Stopping after script 4; you may fill out the file gcode_output.tsv and continue with script 5.\n') #stop = True else: stop = True if stop or args.genetic_code == None: print('\nStopping after script 4 because genetic code information is incomplete. Please fill out the file "gcode_output.tsv" and continue with script 5.\n') exit() def script_five(args): valid_codes = ['bleph','blepharisma','chilo','chilodonella','condy', 'condylostoma','none','eup','euplotes','peritrich','vorticella','ciliate','universal','taa','tag','tga','mesodinium'] lines = [line.strip().split('\t') for line in open(args.output + '/Output/gcode_output.tsv', 'r')] with open(args.output + '/Output/gcode_output.tsv', 'r') as g: for folder in os.listdir(args.output + '/Output'): if os.path.isfile(args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.Renamed.fasta'): for line in lines: if line[0] == folder and line[-1].lower() in valid_codes: os.system('python 5_GCodeTranslate.py --input_file ' + args.output + '/Output/' + folder + '/' + folder + '_WTA_EPU.Renamed.fasta --genetic_code ' + line[-1]) elif line[-1].lower() not in valid_codes and line[-1] != 'Genetic Code': print('\n' + line[-1] + ' is not a valid genetic code. Skipping taxon ' + folder + '.\n') def script_six(args): prefixes = [] for file in os.listdir(args.output + '/Output'): if file.endswith('_AA.ORF.fasta'): prefixes.append(file[:10]) unique_prefixes = list(dict.fromkeys(prefixes)) hook_fasta = '' for file in os.listdir(args.databases + '/db_OG'): if file.split('.')[-1] in ('fasta', 'fas', 'fa', 'faa'): hook_fasta = args.databases + '/db_OG/' + file if hook_fasta == '': print('\nNo .fasta file could be found containing Hook sequences. This should be supplied along with the .dmnd-formatted database file in the Databases/db_OG folder. Quitting before script 6.\n') for prefix in unique_prefixes: os.system('python 6_FilterPartials.py --file_prefix ' + args.output + '/Output/' + prefix + ' --hook_fasta ' + hook_fasta) def script_seven(args): for file in os.listdir(args.output + '/Output/ToRename'): if '.AA.ORF.fasta' in file: os.system('python 7_FinalizeName.py --input_file ' + args.output + '/Output/ToRename/' + file + ' --name ' + file[:10]) os.mkdir(args.output + '/Output/Intermediate') for file in os.listdir(args.output + '/Output'): if file != 'ReadyToGo' and file != 'Intermediate': os.system('mv ' + args.output + '/Output/' + file + ' ' + args.output + '/Output/Intermediate') os.system('python 8_SummaryStats.py -i ' + args.output + '/Output -d ' + args.databases) if __name__ == "__main__": args = get_args() if (args.first_script == 1 or args.script == 1) and (args.assembled_transcripts == None or not os.path.isdir(args.assembled_transcripts)): print('\nERROR: If starting at the first script, a valid path to a folder of assembled transcript files (which must end in .fasta, .fa, or .fna) should be input using the --assembled_transcripts argument') quit() if args.genetic_code == None and args.script == -1: if args.first_script < 5 and args.last_script >= 5: print('\nERROR: You cannot run script 5 without giving a genetic code! If all of the taxa in the run use the same genetic code, then use the --genetic_code argument (e.g. -g Universal). Otherwise, stop after script 4, fill out the spreadsheet called "gcode_translate.tsv," and then run scripts 5-7. If this does not make sense, please ask for help.') quit() ten_digit_codes = [] if args.first_script == 1 or args.script == 1: for file in os.listdir(args.assembled_transcripts): if file[10:] == '_assembledTranscripts.fasta': ten_digit_codes.append(file[:10]) else: if not os.path.isdir(args.output + '/Output'): print('\nERROR: A folder called "Output" is not found at the given output path. Enter the correct path for --output or start from script 1.\n') quit() if(len(ten_digit_codes) > len(list(dict.fromkeys(ten_digit_codes)))): print('\nERROR: Duplicate 10-digit codes are not allowed.\n') quit() for code in ten_digit_codes: for c, char in enumerate(code): if (c != 2 and c != 5 and char not in 'qwertyuiopasdfghjklzxcvbnmQWERTYUIOPASDFGHJKLZXCVBNM1234567890') or ((c == 2 or c == 5) and char != '_'): print('\nERROR: ' + code + ' is an invalid 10-digit code sample identifier. It must of the format Op_me_hsap (Homo sapiens for example). Please ask for help if this does not make sense.\n') quit() if os.path.isdir(args.output + '/Output') and (args.first_script == 1 or args.script == 1): print('\nERROR: An "Output" folder already exists at the given path. Please delete or rename this folder and try again.\n') quit() elif os.path.isdir(args.output + '/Output/Intermediate'): print('\nIt looks like this run is already complete. Try deleting/renaming the Output folder and try again.\n') quit() elif not os.path.isdir(args.output + '/Output'): os.mkdir(args.output + '/Output') scripts = [0, script_one, script_two, script_three, script_four, script_five, script_six, script_seven] if args.script == -1: if args.first_script < args.last_script: for i in range(1 + args.last_script - args.first_script): print('\nRunning script ' + str(i + args.first_script) + '...\n') if i + args.first_script == 1: if len(ten_digit_codes) == 0: print('\nNo properly-named assembled transcripts files found.\n') quit() else: scripts[i + args.first_script](args, ten_digit_codes) else: scripts[i + args.first_script](args) else: print('\nERROR: Invalid script combination: the first script must be less than the last script. If you want to use only once script, use the --script argument.\n') quit() else: if args.script == 1: if len(ten_digit_codes) == 0: print('\nNo properly-named assembled transcripts files found.\n') quit() else: scripts[args.script](args, ten_digit_codes) else: scripts[args.script](args)