Mercurial > repos > card > rgi
changeset 0:715bc9aeef69 draft
planemo upload for repository https://github.com/arpcard/rgi commit 7a78289be23c5a14ae39f454610fa8eca3f05188
| author | card |
|---|---|
| date | Wed, 27 Feb 2019 09:08:21 -0500 |
| parents | |
| children | f96cbb663aa9 |
| files | README.rst data_managers/data_manager_conf.xml data_managers/data_manager_rgi_build_db/data_manager/.gitignore data_managers/data_manager_rgi_build_db/data_manager/import_data.py data_managers/data_manager_rgi_build_db/data_manager/rgi_database_builder.py data_managers/data_manager_rgi_build_db/data_manager/rgi_database_builder.xml data_managers/data_manager_rgi_build_db/tool-data/rgi_databases.loc.sample data_managers/data_manager_rgi_build_db/tool_data_table_conf.xml.sample rgi.xml test-data/test1.fasta |
| diffstat | 10 files changed, 573 insertions(+), 0 deletions(-) [+] |
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--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/README.rst Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,6 @@ +RGI wrapper +-------------------- + +This wrapper is used to run Resistance Gene Identifier (RGI) on galaxy environment. + +This tool is used together with data manager for RGI: `rgi_database_builder <https://github.com/arpcard/rgi_database_builder>`_.
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_conf.xml Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,24 @@ +<?xml version="1.0"?> +<data_managers> + <data_manager tool_file="data_manager_rgi_build_db/data_manager/rgi_database_builder.xml" id="rgi_database_builder" version="1.0.0"> + <!-- Defines a Data Table to be modified --> + <data_table name="rgi_databases"> + <!-- Handle the output of the Data Manager Tool --> + <output> + <!-- Columns that will be specified by the Data Manager Tool --> + <column name="value" /> + <column name="name" /> + <!-- The value of this column will be modified based upon data in "out_file" --> + <column name="path" output_ref="out_file" > + <!-- Moving a file from the extra files path of "out_file" --> + <move type="directory" relativize_symlinks="True"> + <target base="${GALAXY_DATA_MANAGER_DATA_PATH}">rgi_databases/${value}/${path}</target> + </move> + <!-- Store this value in the final Data Table --> + <value_translation>${GALAXY_DATA_MANAGER_DATA_PATH}/rgi_databases/${value}/${path}</value_translation> + <value_translation type="function">abspath</value_translation> + </column> + </output> + </data_table> + </data_manager> +</data_managers>
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_rgi_build_db/data_manager/.gitignore Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,1 @@ +*.pyc \ No newline at end of file
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_rgi_build_db/data_manager/import_data.py Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,199 @@ +import argparse +import datetime +import json +import os +import shutil +import sys +import tarfile +import urllib.request, urllib.error, urllib.parse +import zipfile +import logging + +path = os.path.join(os.getcwd(), 'rgi-database') +data_path = path + +level = logging.WARNING +logger = logging.getLogger(__name__) +logger.setLevel(level) + +def url_download(url, workdir): + file_path = os.path.join(workdir, 'download.dat') + if not os.path.exists(workdir): + os.makedirs(workdir) + src = None + dst = None + try: + req = urllib.request.Request(url) + src = urllib.request.urlopen(req) + dst = open(file_path, 'wb') + while True: + chunk = src.read(2**10) + if chunk: + dst.write(chunk) + else: + break + except Exception as e: + print(str(e), file=sys.stderr) + finally: + if src: + src.close() + if dst: + dst.close() + if tarfile.is_tarfile(file_path): + fh = tarfile.open(file_path, 'r:*') + elif zipfile.is_zipfile(file_path): + fh = zipfile.ZipFile(file_path, 'r') + else: + return + # extract only one file : card.json + for member in fh.getmembers(): + if member.isreg(): # skip if the TarInfo is not files + member.name = os.path.basename(member.name) # remove the path by reset it + if member.name == 'card.json': + print('[import_data] extracting file: {}'.format(str(member.name))) + fh.extract(member.name,workdir) + os.remove(file_path) + +def checkKeyExisted(key, my_dict): + try: + nonNone = my_dict[key] is not None + except KeyError: + nonNone = False + return nonNone + +def data_version(): + data_version = '' + with open(os.path.join(data_path, 'card.json')) as json_file: + json_data = json.load(json_file) + for item in list(json_data.keys()): + if item == '_version': + data_version = json_data[item] + json_file.close() + return data_version + +def makeBlastDB(): + if os.path.isfile(os.path.join(path, 'proteindb.fsa')) == True: + print('[import_data] create blast DB.') + os.system('makeblastdb -in {}/proteindb.fsa -dbtype prot -out {}/protein.db > /dev/null 2>&1'.format(path, path)) + +def makeDiamondDB(): + if os.path.isfile(os.path.join(path, 'proteindb.fsa')) == True: + print('[import_data] create diamond DB.') + os.system('diamond makedb --quiet --in {}/proteindb.fsa --db {}/protein.db'.format(path, path)) + +def write_fasta_from_json(): + '''Creates a fasta file from card.json file.''' + if os.path.isfile(os.path.join(path, 'proteindb.fsa')): + return + else: + try: + with open(os.path.join(data_path, 'card.json'), 'r') as jfile: + j = json.load(jfile) + except Exception as e: + logger.error(e) + exit() + + with open(os.path.join(path, 'proteindb.fsa'), 'w') as fout: + for i in j: + if i.isdigit(): + # model_type: protein homolog model + if j[i]['model_type_id'] == '40292': + try: + pass_bit_score = j[i]['model_param']['blastp_bit_score']['param_value'] + except KeyError: + logger.warning('No bitscore for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + else: + try: + for seq in j[i]['model_sequences']['sequence']: + fout.write('>%s_%s | model_type_id: 40292 | pass_bitscore: %s | %s\n' % (i, seq, pass_bit_score, j[i]['ARO_name'])) + fout.write('%s\n' %(j[i]['model_sequences']['sequence'][seq]['protein_sequence']['sequence'])) + except Exception as e: + logger.warning('No model sequences for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + + + # model_type: protein variant model + elif j[i]['model_type_id'] == '40293': + try: + pass_bit_score = j[i]['model_param']['blastp_bit_score']['param_value'] + except KeyError: + logger.warning('No bitscore for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + else: + try: + snpList = [j[i]['model_param']['snp']['param_value'][k] for k in j[i]['model_param']['snp']['param_value']] + except Exception as e: + logger.warning('No snp for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + + try: + for seq in j[i]['model_sequences']['sequence']: + fout.write('>%s_%s | model_type_id: 40293 | pass_bit_score: %s | SNP: %s | %s\n' \ + % (i, seq, pass_bit_score, ','.join(snpList), j[i]['ARO_name'])) + fout.write('%s\n' % (j[i]['model_sequences']['sequence'][seq]['protein_sequence']['sequence'])) + except Exception as e: + logger.warning('No model sequences for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + + # model_type: protein overexpression model + elif j[i]['model_type_id'] == '41091': + try: + pass_bit_score = j[i]['model_param']['blastp_bit_score']['param_value'] + except KeyError: + logger.warning('No bitscore for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + else: + try: + snpList = [j[i]['model_param']['snp']['param_value'][k] for k in j[i]['model_param']['snp']['param_value']] + except Exception as e: + logger.warning('No snp for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + + try: + for seq in j[i]['model_sequences']['sequence']: + fout.write('>%s_%s | model_type_id: 41091 | pass_bit_score: %s | SNP: %s | %s\n' \ + % (i, seq, pass_bit_score, ','.join(snpList), j[i]['ARO_name'])) + fout.write('%s\n' % (j[i]['model_sequences']['sequence'][seq]['protein_sequence']['sequence'])) + except Exception as e: + logger.warning('No model sequences for model (%s, %s). RGI will omit this model and keep running.' \ + % (j[i]['model_id'], j[i]['model_name'])) + logger.info('Please let the CARD Admins know! Email: card@mcmaster.ca') + +def _main(args): + if not os.path.exists(path): + print('[import_data] mkdir: {}'.format(path)) + os.makedirs(path) + print('[import_data] path: {}'.format(path)) + print(args) + + if args.url == None: + url = 'https://card.mcmaster.ca/latest/data' + else: + url = args.url + print('[import_data] url: {}'.format(url)) + workdir = os.path.join(os.getcwd(), 'rgi-database') + print('[import_data] working directory: {}'.format(workdir)) + url_download(url, workdir) + write_fasta_from_json() + makeBlastDB() + makeDiamondDB() + version = data_version() + print('[import_data] data version: {}'.format(version)) + return version + +def run(): + parser = argparse.ArgumentParser(description='Create data manager json.') + parser.add_argument('--url', dest='url', action='store', help='Url for CARD data') + args = parser.parse_args() + _main(args) + +if __name__ == '__main__': + run()
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_rgi_build_db/data_manager/rgi_database_builder.py Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,47 @@ +import argparse +import datetime +import json +import os +import shutil +import sys +import tarfile +import urllib.request, urllib.error, urllib.parse +import zipfile +from import_data import run, _main + +parser = argparse.ArgumentParser(description='Create data manager json.') +parser.add_argument('--url', dest='url', action='store', help='Url for CARD data') +parser.add_argument('--out', dest='output', action='store', help='JSON filename') +parser.add_argument('--name', dest='name', action='store', default='CARD_data-' + str(datetime.datetime.now().strftime('%Y-%B-%d-%H:%M:%S')), help='Data table database name') +args = parser.parse_args() + +print('[rgi_database_builder] Importing...') + +_main(args) + +def main(args): + print('[rgi_database_builder] Building......') + + data_manager_entry = {} + data_manager_entry['value'] = args.name.lower() + data_manager_entry['name'] = args.name + data_manager_entry['path'] = '.' + + data_manager_json = dict(data_tables = dict(rgi_databases=data_manager_entry)) + + params = json.loads(open(args.output,'r').read()) + + target_directory = params['output_data'][0]['extra_files_path'] + os.mkdir(target_directory) + output_path = os.path.join(os.getcwd(), 'rgi-database') + + for filename in os.listdir(output_path): + print('[rgi_database_builder] move file: {} from {} to {}'.format(filename, output_path, target_directory)) + shutil.move(os.path.join(output_path, filename), target_directory) + + print(args.output) + print('[rgi_database_builder] write file: {}'.format(args.output)) + open(args.output, 'w').write(json.dumps(data_manager_json)) + +if __name__ == '__main__': + main(args)
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_rgi_build_db/data_manager/rgi_database_builder.xml Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,28 @@ +<?xml version="1.0"?> +<tool id="rgi_database_builder" name="RGI Database Builder" tool_type="manage_data" version="1.0.0"> + <description>Download and build the CARD database for RGI</description> + <requirements> + <requirement type="package" version="4.2.2">rgi</requirement> + </requirements> + <stdio> + <exit_code description="Error" level="fatal" range="1:" /> + </stdio> + <command interpreter="python3"> + <![CDATA[ + rgi_database_builder.py --out "${out_file}" + #if $name: + --name "${name}" + #end if + #if $url: + --url "${url}" + #end if + ]]> + </command> + <inputs> + <param name="name" type="text" value="" label="Database name" help="Name for this database, or leave blank for today's date" optional="True"/> + <param name="url" type="text" value="https://card.mcmaster.ca/latest/data" label="Url" help="Url for the CARD data e.g https://card.mcmaster.ca/lastest/data. Visit https://card.mcmaster.ca/download to see previous CARD releases." optional="True"/> + </inputs> + <outputs> + <data format="data_manager_json" name="out_file" label="${tool.name} (JSON)"/> + </outputs> +</tool>
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_rgi_build_db/tool-data/rgi_databases.loc.sample Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,5 @@ +#This file lists the columns that will be specified by the RGI Data Manager tool. +# +#For example: +#<value> <name> <path> +#rgi_20181001 rgi_20181001 /galaxy-central/tool-data/rgi_databases/rgi_20181001
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/data_managers/data_manager_rgi_build_db/tool_data_table_conf.xml.sample Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,8 @@ +<?xml version="1.0"?> +<tables> + <!-- Locations of RGI database in the required format --> + <table name="rgi_databases" comment_char="#"> + <columns>value, name, path</columns> + <file path="tool-data/rgi_databases.loc" /> + </table> +</tables>
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/rgi.xml Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,187 @@ +<tool id="rgi" name="Resistance Gene Identifier (RGI)" version="4.2.2"> + <description>This tool predicts resistome(s) from protein or nucleotide data based on homology and SNP models.</description> + <requirements> + <requirement type="package" version="4.2.2">rgi</requirement> + </requirements> + <stdio> + <exit_code range="1:" /> + </stdio> + <command><![CDATA[ +#if $db_opts.db_opts_selector != "default": + rgi load + #if $db_opts.db_opts_selector == "local": + -i "${__tool_data_path__}/rgi_databases/$db_opts.rgi_db_local/card.json" + #else: + -i "$db_opts.rgi_db_hist" + #end if + --local + && +#end if + +rgi main + -i '$input_sequence' + -o out + -t $input_type + -a $alignment_tool + -n "\${GALAXY_SLOTS:-1}" + $include_loose + $low_quality + -d $data + +#if $db_opts.db_opts_selector != "default": + --local +#end if + ]]></command> + <inputs> + <param argument="--input_sequence" type="data" format="fastq,fasta" label="Input sequence" help="input file must be in either FASTA (contig and protein) or gzip format e.g myFile.fasta, myFasta.fasta.gz"/> + <param argument="--input_type" type="select" multiple="false" label="Input type" help="specify data input type (default = contig)"> + <option value="contig" selected="true">CONTIG (Nucleotide Sequence)</option> + <option value="protein">PROTEIN (Protein Sequence)</option> + </param> + <param argument="--alignment_tool" type="select" label="Alignment tool" help="specify alignment tool (default = BLAST)"> + <option value="blast" selected="true">BLAST</option> + <option value="diamond">DIAMOND</option> + </param> + <param argument="--include_loose" type="boolean" checked="false" truevalue="--include_loose" falsevalue="" label="Include loose hits" help="include loose hits in addition to strict and perfect hits"/> + <param argument="--low_quality" type="boolean" checked="false" truevalue="--low_quality" falsevalue="" label="Low quality" help="use for short contigs to predict partial genes"/> + <param argument="--data" type="select" multiple="false" label="Data type" help="specify a data-type (default = NA)"> + <option value="wgs">WGS</option> + <option value="plasmid">PLASMID</option> + <option value="chromosome">CHROMOSOME</option> + <option value="NA" selected="true">NA</option> + </param> + <conditional name="db_opts"> + <param name="db_opts_selector" type="select" label="Select an RGI database"> + <option value="default" selected="true">Default RGI database</option> + <option value="local">Locally installed RGI database</option> + <option value="hist">RGI database from your history</option> + </param> + <when value="default"> + <param name="rgi_db_local" type="hidden" value="" /> + <param name="rgi_db_hist" type="hidden" value="" /> + </when> + <when value="local"> + <param name="rgi_db_local" type="select" multiple="false" label="Locally installed RGI database"> + <options from_data_table="rgi_databases"> + <validator type="no_options" message="No databases are available built-in"/> + </options> + </param> + <param name="rgi_db_hist" type="hidden" value="" /> + </when> + <when value="hist"> + <param name="rgi_db_local" type="hidden" value="" /> + <param name="rgi_db_hist" type="data" format="json" label="RGI database from your history" /> + </when> + </conditional> + </inputs> + <outputs> + <data name="report" format="json" from_work_dir="out.json" label="${tool.name} on ${on_string}: report.json"/> + <data name="summary" format="tabular" from_work_dir="out.txt" label="${tool.name} on ${on_string}: summary.txt"/> + </outputs> + <tests> + <test> + <param name="input_sequence" value="test1.fasta" /> + <param name="input_type" value="contig" /> + <param name="alignment_tool" value="blast" /> + <param name="include_loose" value="false" /> + <param name="low_quality" value="false" /> + <param name="data" value="NA" /> + <conditional name="db_opts"> + <param name="db_opts_selector" value="default" /> + </conditional> + <output name="summary"> + <assert_contents> + <has_text text="AY123251" /> + </assert_contents> + </output> + </test> + </tests> + <help> + <![CDATA[ +Resistance Gene Identifier (RGI) +================================ + +RGI is used to predict resistome(s) from protein or nucleotide data based on homology and SNP models. The tool uses data from the CARD database. + +Usage +----- + +1. Select your input sequence (in FASTA format). +2. Select your input type (CONTIG or PROTEIN). +3. Select your alignment tool (DIAMOND or BLAST). +4. Specify if you want to include loose hits (YES or NO). +5. Specify if you want to remove temporary files (YES or NO). +6. Specify if you want to low quality predictions (YES or NO). +7. Select your data type (WGS, PLASMID, CHROMOSOME or NA). +8. Run the tool. + +Output +------ + +There are 2 different output files produced by `rgi`. + +summary.txt +^^^^^^^^^^^ + +A tabular file of all detected resistance genes, one gene per line. + ++--------------------------------------------------------------------------------------------------------------------------+---------------+-------+------+-------------+----------+----------------+---------------------+----------------+------------------+----------+-----------------------+--------------------------+-------------+------------+-------------------------+----------------------+---------------------------+------------------------+---------------------------+-----------------------------------------+--------------------------------+-----------+ +| ORF_ID | Contig | Start | Stop | Orientation | Cut_Off | Pass_Bitscore | Best_Hit_Bitscore | Best_Hit_ARO | Best_Identities | ARO | Model_type | SNPs_in_Best_Hit_ARO | Other_SNPs | Drug Class | Resistance Mechanism | AMR Gene Family | Predicted_DNA | Predicted_Protein | CARD_Protein_Sequence | Percentage Length of Reference Sequence | ID | Model_ID | ++==========================================================================================================================+===============+=======+======+=============+==========+================+=====================+================+==================+==========+=======================+==========================+=============+============+=========================+======================+===========================+========================+===========================+=========================================+================================+===========+ +| AY123251.1_6 # 3575 # 4489 # 1 # ID=1_6;partial=00;start_type=ATG;rbs_motif=None;rbs_spacer=None;gc_cont=0.399 | AY123251.1_6 | 3575 | 4489 | \+ | Strict | 500 | 506.908 | CARB-7 | 83.68 | 3002246 | protein homolog model | n/a | n/a | penam | antibiotic inactivation | CARB beta-lactamase | ATGC | MLLYKMCDNQNYGVTYMKFLLV | MKSLLVFALLMPSVVFASSSKFQSV | 105.56 | gnl|BL_ORD_ID|589|hsp_num:0 | 1443 | ++--------------------------------------------------------------------------------------------------------------------------+---------------+-------+------+-------------+----------+----------------+---------------------+----------------+------------------+----------+-----------------------+--------------------------+-------------+------------+-------------------------+----------------------+---------------------------+------------------------+---------------------------+-----------------------------------------+--------------------------------+-----------+ + +report.json +^^^^^^^^^^^ + +A json version of summary.txt. + +Help +---- + +**usage:** + ``rgi main [-h] -i INPUT_SEQUENCE -o OUTPUT_FILE [-t {read,contig,protein,wgs}] [-a {DIAMOND,BLAST}] [-n THREADS] [--include_loose] [--local] [--clean] [--debug] [--low_quality] [-d {wgs,plasmid,chromosome,NA}] [-v]`` + +**optional arguments:** + +- ``-h, --help`` + show this help message and exit +- ``-i INPUT_SEQUENCE, --input_sequence INPUT_SEQUENCE`` + input file must be in either FASTA (contig and protein) or gzip format e.g myFile.fasta, myFasta.fasta.gz +- ``-o OUTPUT_FILE, --output_file OUTPUT_FILE`` + output folder and base filename +- ``-t {read,contig,protein,wgs}, --input_type {read,contig,protein,wgs}`` + specify data input type (default = contig) +- ``-a {DIAMOND,BLAST}, --alignment_tool {DIAMOND,BLAST}`` + specify alignment tool (default = BLAST) +- ``-n THREADS, --num_threads THREADS`` + number of threads (CPUs) to use in the BLAST search (default=8) +- ``--include_loose`` + include loose hits in addition to strict and perfect hits +- ``--local`` + use local database (default: uses database in executable directory) +- ``--clean`` + removes temporary files +- ``--debug`` + debug mode +- ``--low_quality`` + use for short contigs to predict partial genes +- ``-d {wgs,plasmid,chromosome,NA}, --data {wgs,plasmid,chromosome,NA}`` + specify a data-type (default = NA) +- ``-v, --version`` + prints software version number + +Links +----- + +RGI: https://card.mcmaster.ca/analyze/rgi + +Github: https://github.com/arpcard/rgi + +CARD database: https://card.mcmaster.ca + ]]> + </help> + <citations> + <citation type="doi">10.1093/nar/gkw1004</citation> + </citations> +</tool>
--- /dev/null Thu Jan 01 00:00:00 1970 +0000 +++ b/test-data/test1.fasta Wed Feb 27 09:08:21 2019 -0500 @@ -0,0 +1,68 @@ +>AY123251.1 Salmonella enterica subsp. enterica serovar Typhi R-plasmid pST2301 class I integron aminoglycoside 6'-N-acetyltransferase (aacA4), chloramphenicol acetyltransferase (catB8), aminoglycoside 3'-adenyltransferase (aadA1), dihydrofolate reductase type I (dhfr1), aminoglycoside 6'-n-acetyltransferase (aac6-II), and CARB-8 beta-lactamase (blaCARB-8) genes, complete cds +TCATGGCTTGTTATGACTGTTTTTTTGTACAGTCTATGCCTCGGGCATCCAAGCAGCAAGCGCGTTACGC +CGTGGGTCGATGTTTGATGTTATGGAGCAGCAACGATGTTACGCAGCAGGGCAGTCGCCCTAAAACAAAG +TTAGGCATCACAAAGTACAGCATCGTGACCAACAGCAACGATTCCGTCACACTGCGCCTCATGACTGAGC +ATGACCTTGCGATGCTCTATGAGTGGCTAAATCGATCTCATATCGTCGAGTGGTGGGGCGGAGAAGAAGC +ACGCCCGACACTTGCTGACGTACAGGAACAGTACTTGCCAAGCGTTTTAGCGCAAGAGTCCGTCACTCCA +TACATTGCAATGCTGAATGGAGAGCCGATTGGGTATGCCCAGTCGTACGTTGCTCTTGGAAGCGGGGACG +GATGGTGGGAAGAAGAAACCGATCCAGGAGTACGCGGAATAGACCAGTCACTGGCGAATGCATCACAACT +GGGCAAAGGCTTGGGAACCAAGCTGGTTCGAGCTCTGGTTGAGTTGCTGTTCAATGATCCCGAGGTCACC +AAGATCCAAACGGACCCGTCGCCGAGCAACTTGCGAGCGATCCGATGCTACGAGAAAGCGGGGTTTGAGA +GGCAAGGTACCGTAACCACCCCAGATGGTCCAGCCGTGTACATGGTTCAAACACGCCAGGCATTCGAGCG +AACACGCAGTGTTGCCTAACCCTTCCATCGAGGGGGACGTCCAAGGGCTGGCGCCCTTGGCCGCCCCTCA +TGTCAAACGTTAGACGGCAAGAAAAGGTTCCACGAACTCTGATGAAAAACTACTTTAACAGCCCTTTCAA +AGGGGAACTTCTTTCTGAGCAAGTGAAAAATCCAAATATCAGAGTAGGCCGGTATAGCTATTACTCTGGC +TACTATCACGGGCACTCATTTGATGAATGCGCGCGATACTTGCTTCCAGATCGTGATGACGTTGATAAAT +TGATCATTGGCAGCTTTTGTTCTATAGGAAGCGGGGCTTCCTTCATCATGGCTGGCAATCAGGGGCATCG +GCATGACTGGGCATCATCCTTCCCCTTCTTCTATATGCAAGAGGAGCCTGCTTTCTCAAGAGCACTCGAC +GCCTTCCAAAGAGCAGGTGATACCGTCATTGGCAATGATGTCTGGATAGGCTCGGAGGCAATGATTATGC +CTGGCATCAAAATTGGAGACGGTGCCGTGATAGGTAGTCGCTCGTTGGTGACAAAAGATGTAGAGCCTTA +TGCCATCATCGGGGGAAATCCCGCAAAGCAAATTAAGAAGCGCTTCTCCGATGAGGAAATCTCATTGCTC +ATGGAGATGGAGTGGTGGAACTGGCCACTAGATAAAATTAAGACAGCAATGCCTCTGCTGTGCTCGTCAA +ATATTTTTGGTCTGCATAAGTATTGGCGCGAGTTTGCCGTCTAACAATTCATTCAAGCCGACGCCGCTTC +GCGGCACGGCTTAATTCTGGCGTTAAACATCATGAGGGAAGCGGTGATCGCCGAAGTATCGACTCAACTA +TCAGAGGTAGTTGGCGTCATCGAGCGCCATCTCGAACCGACGTTGCTGGCCGTACATTTGTACGGCTCCG +CAGTGGATGGCGGCCTGAAGCCACACAGTGATATTGATTTGCTGGTTACGGTGACCGTAAGGCTTGATGA +AACAACGCGGCGAGCTTTGATCAACGACCTTTTGGAAACTTCGGCTTCCCCTGGAGAGAGCGAGATTCTC +CGCGCTGTAGAAGTCACCATTGTTGTGCACGACGACATCATTCCGTGGCGTTATCCAGCTAAGCGCGAAC +TGCAATTTGGAGAATGGCAGCGCAATGACATTCTTGCAGGTATCTTCGAGCCAGCCACGATCGACATTGA +TCTGGCTATCTTGCTGACAAAAGCAAGAGAACATAGCGTTGCCTTGGTAGGTCCAGCGGCGGAGGAACTC +TTTGATCCGGTTCCTGAACAGGATCTATTTGAGGCGCTAAATGAAACCTTAACGCTATGGAACTCGCCGC +CCGACTGGGCTGGCGATGAGCGAAATGTAGTGCTTACGTTGTCCCGCATTTGGTACAGCGCAGTAACCGG +CAAAATCGCGCCGAAGGATGTCGCTGCCGACTGGGCAATGGAGCGCCTGCCGGCCCAGTATCAGCCCGTC +ATACTTGAAGCTAGACAGGCTTATCTTGGACAAGAAGAAGATCGCTTGGCCTCCCGCGCAGATCAGTTGG +AAGAATTTGTTCACTACGTGAAAGGCGAGATCACCAAGGTAGTCGGCAAATAATGTCTAACAATTCGTTC +AAGCCGACGCCGCTTCGCGGCGCGGCTTAACTCAAGCGTTAACCTCTGAGGAAGAATTGTGAAACTATCA +CTAATGGTAGCTATATCGAAGAATGGAGTTATCGGGAATGGCCCTGATATTCCATGGAGTGCCAAAGGTG +AACAGCTCCTGTTTAAAGCTATTACCTATAACCAATGGCTGTTGGTTGGACGCAAGACTTTTGAGTCAAT +GGGAGCATTACCCAACCGAAAGTATGCGGTCGTAACACGTTCAAGTTTTACATCTGACAATGAGAACGTA +GTGATCTTTCCATCAATTAAAGATGCTTTAACCAACCTAAAGAAAATAACGGATCATGTCATTGTTTCAG +GTGGTGGGGAGATATACAAAAGCCTGATCGATCAAGTAGATACACTACATATATCTACAATAGACATCGA +GCCGGAAGGTGATGTTTACTTTCCTGAAATCCCCAGCAATTTTAGGCCAGTTTTTACCCAAGACTTCGCC +TCTAACATAAATTATAGTTACCAAATCTGGCAAAAGGGTTAACAAGTGGCAGCAACGGATTCGCAAACCT +GTCACGCCTTTTGTACCAAAACCCGCGCCAGGTTTGCGATCCGCTGTGCCAGGCGTTAGGCAGCACAGAG +CGACCATTTCATGTCCGCGAGCACCCCCCCCATAACTCTTCGCCTCATGACCGAGCGCGACCTGCCGATG +CTCCATGATTGGCTCAACCGGCCGCACATCGTTGAGTGGTGGGGTGGTGACGAAGAGCGACCGACTCTTG +ATGAAGTGCTGGAACACTACCTGCCCAGAGCGATGGCGGAAGAGTCCGTAACACCGTACATCGCAATGCT +GGGCGAGGAACCGATCGGCTATGCTCAGTCGTACGTCGCGCTCGGAAGCGGTGATGGCTGGTGGGAAGAT +GAAACTGATCCAGGAGTGCGAGGAATAGACCAGTCTCTGGCTGACCCGACACAGTTGAACAAAGGCCTAG +GAACAAGGCTTGTCCGCGCTCTCGTTGAACTACTGTTCTCGGACCCCACCGTGACGAAGATTCAGACCGA +CCCGACTCCGAACAACCATCGAGCCATACGCTGCTATGAGAAGGCAGGATTCGTGCGGGAGAAGATCATC +ACCACGCCTGACGGGCCGGCGGTTTACATGGTTCAAACACGACAAGCCTTCGAGAGAAAGCGCGGTGTTG +CCTAACAACTCATTCAAGCCGACGCCGCTTCGCGGCGCGGCTTAATTCAGGTGTTAGCCATATTATGGAG +CCTCATGCTTTTATATAAAATGTGTGACAATCAAAATTATGGGGTTACTTACATGAAGTTTTTATTGGTA +TTTTCGCTTTTAATACCATCCGTGGTTTTTGCAAGTAGTTCAAAGTTTCGGCAAGTTGAACAAGACGTTA +AGGCAATTGAAGTTTCTCTTTCTGCTCGTATAGGTGTTTCCGTTCTTGATACTCAAAATGGAGAATACTG +GGATTACAATGGCAATCAGCGCTTCCCGTTGACAAGTACTTTTAAAACAATAGCTTGCGCTAAATTACTA +TATGATGCTGAGCAAGGAAAAGTTAATCCCAATAGTACAATCGAGATTAAGAAAGCAGATCTTGTGACCT +ATTCCCCTGTAATAGAAAAGCAAGTAGGGCAGGCAATCACACTCGATGATGCGTGCTTCGCAACTATGAC +TACAAGTGATAATACTGCGGCAAATATCATCCTAAGTGCTGTAGGTGGCTCCAAAGGCGTTACTGATTTT +TTAAGACAAATTGGGGACAAAGAGACTCGTCTAGACCGTATTGAGCCTGATTTAAATGAAGGTAAGCTCG +GTGATTTGAGGGATACGACAACTCCTAAGGCAATAGCCAGTACTTTGAATAAATTTTTATTTGGTTCAGC +GCTATCTGAAATGAACAAAAAAAAATTAGAGTCTTGGATGGTGAACAATCAAGTCACTGGTAATTTACTA +CGTTCAGTATTGCCGGCGGGATGGAACATTGCGGATCGTTCAGGTGCTGGCGGATTTGGTGCTCGGAGTA +TTACAGCAGTTGTGTGGAGTGAGCATCAAGCCCCAATTATTGTGAGCATCTATCTAGCTCAAACACAGGC +TTCAATGGCAGAGCGAAATGATGCGATTGTTAAAATTGGTCGTTCAATTTTTGACGTTTATACATCACAG +TCGCGCTGATAAGGCTAACAAGGCCATCAAGTTGACGGCTTTTCCGTCGCTTGTTTTGTGGCTTAACGCT +ACGCTACCACAAAACAATCAACTACAAAGCCGCAACTTATGGCGGCGTTAGATACACTAAGCACATAATT +GCTCACAGCCAAACTATCAGGTCAAGTCTGCTTTTATTATTTTTAAGCGTGCATAATAAGCCCTAC \ No newline at end of file