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changeset 5:7a7d7f30ddfc draft

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"planemo upload for repository https://github.com/galaxyproject/tools-iuc/tree/master/tools/artic commit dacdfa7ecfbdd1c1a12494cf7d0450185c122c84"
author iuc
date Fri, 27 Aug 2021 20:34:32 +0000
parents a346730e5552
children f212134e204c
files artic_minion.xml macros.xml test-data/gupplyplex_output.fastq test-data/test.fastq
diffstat 4 files changed, 126 insertions(+), 14 deletions(-) [+]
line wrap: on
line diff
--- a/artic_minion.xml	Fri Oct 09 17:47:34 2020 +0000
+++ b/artic_minion.xml	Fri Aug 27 20:34:32 2021 +0000
@@ -1,21 +1,24 @@
-<tool id="artic_minion" name="ARTIC minion" version="1.1.3+galaxy2">
+<tool id="artic_minion" name="ARTIC minion" version="@PACKAGE_VERSION@+galaxy0" profile="20.09">
     <description>Build consensus sequence and call variants from amplicon-based nanopore sequence data</description>
+    <macros>
+        <import>macros.xml</import>
+    </macros>
     <requirements>
-        <requirement type="package" version="1.1.3">artic</requirement>
+        <requirement type="package" version="@PACKAGE_VERSION@">artic</requirement>
     </requirements>
     <command detect_errors="exit_code"><![CDATA[
-        mkdir -p 'scheme/name/version' &&
+        mkdir -p 'scheme/name/V1' &&
         #if str( $primer_scheme_source.primer_scheme_source_selector ) == "tool_data_table":
-          ln -s '${primer_scheme_source.primer_scheme_bedfile.fields.path}' 'scheme/name/version/name.scheme.bed' &&
+          ln -s '${primer_scheme_source.primer_scheme_bedfile.fields.path}' 'scheme/name/V1/name.scheme.bed' &&
         #else:
-          ln -s '${primer_scheme_source.primer_scheme_bedfile}' 'scheme/name/version/name.scheme.bed' &&
+          ln -s '${primer_scheme_source.primer_scheme_bedfile}' 'scheme/name/V1/name.scheme.bed' &&
         #end if
         #if str( $reference_source.reference_source_selector ) == "history":
-          ln -s '${reference_source.reference}' 'scheme/name/version/name.reference.fasta' &&
-          samtools faidx 'scheme/name/version/name.reference.fasta' &&
+          ln -s '${reference_source.reference}' 'scheme/name/V1/name.reference.fasta' &&
+          samtools faidx 'scheme/name/V1/name.reference.fasta' &&
         #else:
-          ln -s '${reference_source.reference.fields.path}' 'scheme/name/version/name.reference.fasta' &&
-          samtools faidx 'scheme/name/version/name.reference.fasta' &&
+          ln -s '${reference_source.reference.fields.path}' 'scheme/name/V1/name.reference.fasta' &&
+          samtools faidx 'scheme/name/V1/name.reference.fasta' &&
         #end if
         artic minion
             --threads \${GALAXY_SLOTS:-1}
@@ -25,8 +28,9 @@
             --read-file '${read_file}'
             --scheme-directory 'scheme'
             --medaka
+            --medaka-model '$medaka_model'
             $bwa
-            'name/version'
+            'name/V1'
             '${read_file.element_identifier}'
         && bgzip -f '${read_file.element_identifier}.fail.vcf'
     ]]></command>
@@ -71,6 +75,9 @@
                        help="You can upload a FASTA sequence to the history and use it as reference" />
             </when>
         </conditional>
+        <param type="text" name="medaka_model" label="Medaka model" help="Model string to pass to medaka (see https://github.com/nanoporetech/medaka#models)">
+            <validator type="expression" message="Please specify a valid medaka model string (see https://github.com/nanoporetech/medaka#models)">(len(value.strip().split('_')) == 3 or len(value.strip().split('_')) == 4) and value.strip().startswith('r')</validator>
+        </param>
     </inputs>
     <outputs>
         <data name="alignment_trimmed" format="bam" from_work_dir="*.primertrimmed.rg.sorted.bam" label="${tool.name} on ${on_string}: trimmed alignment" />
@@ -89,6 +96,7 @@
             <param name="reference" value="nCoV-2019.reference.fasta" />
             <param name="primer_scheme_source_selector" value="tool_data_table" />
             <param name="primer_scheme_bedfile" value="test_entry" />
+            <param name="medaka_model" value="r941_min_high_g360" />
             <output name="consensus_fasta" file="SRR11410539_seqtk_sample_500_1.fastq.consensus.fasta" />
         </test>
         <test>
@@ -97,6 +105,7 @@
             <param name="reference" value="nCoV-2019.reference.fasta" />
             <param name="primer_scheme_source_selector" value="history" />
             <param name="primer_scheme_bedfile" value="nCoV-2019.scheme.V1.bed" />
+            <param name="medaka_model" value="r941_min_high_g360" />
             <output name="consensus_fasta" file="SRR11410539_seqtk_sample_500_1.fastq.consensus.fasta" />
         </test>
         <test>
@@ -105,15 +114,28 @@
             <param name="reference" value="test_entry" />
             <param name="primer_scheme_source_selector" value="tool_data_table" />
             <param name="primer_scheme_bedfile" value="test_entry" />
+            <param name="medaka_model" value="r941_min_high_g360" />
             <output name="consensus_fasta" file="SRR11410539_seqtk_sample_500_1.fastq.consensus.fasta" />
         </test>
     </tests>
     <help><![CDATA[
-This tool aligns MinION reads that were generated from a tiling amplicon library against a viral reference sequence. 
+ARTIC_ minion aligns Nanopore reads that were generated from a tiling amplicon library against a viral reference sequence. 
 It generates a consensus fasta file and a vcf variant file.
 
-This tool is configured to use the experimental 'medaka' variant caller.
+This tool is configured to use the experimental 'medaka' variant caller and must be supplied with the name of
+a model file to use with 'medaka', see the `medaka web page`_ for details.
+
+.. _ARTIC: https://artic.readthedocs.io/en/latest/
+.. _medaka web page: https://github.com/nanoporetech/medaka#models
+
+Note that you should choose an appropriate model for the medaka version used by ARTIC minion.
+
+==================  ==================
+  ARTIC version     medaka version
+==================  ==================
+ 1.2.1                1.0.3
+ 1.3.0-dev            1.2.3
+==================  ==================
     ]]></help>
-    <citations>
-    </citations>
+    <expand macro="citations" />
 </tool>
--- /dev/null	Thu Jan 01 00:00:00 1970 +0000
+++ b/macros.xml	Fri Aug 27 20:34:32 2021 +0000
@@ -0,0 +1,18 @@
+<macros>
+    <token name="@PACKAGE_VERSION@">1.2.1</token>
+    <xml name="citations">
+        <citations>
+            <citation type="bibtex">
+                @misc{githubartic-ncov2019,
+                  author = {Nick Loman and the ZiBRA Project and the ARTIC project},
+                  year = {2017},
+                  title = {ARTIC toolkit},
+                  publisher = {GitHub},
+                  journal = {GitHub repository},
+                  howpublished{\url{https://github.com/artic-network/fieldbioinformatics}},
+                }
+                </citation>
+                <yield />
+        </citations>
+    </xml>
+</macros>
--- /dev/null	Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/gupplyplex_output.fastq	Fri Aug 27 20:34:32 2021 +0000
@@ -0,0 +1,32 @@
+@9bd6c9e7-e630-471b-88f5-f0f46e5b3767
+CATCGGTGTTGGCCCGTTTCAGTTACGTAACCTACCAACCAGCTTTCGATCTCTTGTAGATCTATTCTCTAAACGAACTTTAAAATCTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAACTAGTACTGTCGTCTGACAGGACGAAATAACTCGTCTATCTTCTGCAGGCTGTTACGGTTTTCGTCCGTGTACGGCCGATCATCAACACATCTAGGTTTGTCCGGGTGTGACGAAAATGAATGGAAGCCTTGTCCCCTGGTTTCAACGAGAAAACACGTCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCATGCGTGGCTTTGGAACTCCGTGGAGGTCTTATCAGGGCATCAGCATGTAAAGATGAGCAATACGTAACT
++
+'%*+%%&#$%%%((*,88):*-++*,'$$#$%$#++13/*(%$$&/7%'13)034)--2/,7'$798<@8:<D568;@BDCLG216A88#)-;6..1%##''))++$$--)*+&(.,,.>>A2/0//1124664;67:>6014*8315---&'%%%%%%.+$%%)*%'''(*.%''37A8I<87*.,/552)$&%%$&=AB>9200402)&%'&)454?;>--8+'),)/8B/0C:=HB1((>&+030DE,6'01**))'+&&'''.0,020//4.&0(*7?A0/.023(2.42-+.(%=1.;:A3'A:)976;.LLP*%(,%2@=61.0&,,4/?D=&&&)(...$$,7)-,'<<B57///+,4888@344&(&'$$.?2*./(%)//01<''%5;;:<<=6231%%
+@2912dba0-4acb-4b30-aa79-115c7c6e3c6c
+CGATGTACTTCGTTCAGTTACGTAACACTACCAACCAACTTTCGATCTACCCGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCCGGCTGCATCGCCCAGTGCACTCACCAAGTATAATTAATAACTAATTACTGTCGTTGACAGGTTTTCGAGTAACTCTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTTACGACCGATCATCAACATCTATGATGGTTTTGTCAGGTGTGACCGAAAGGTAAGATGGAGAGCCTTGTCCCCCTGGTTTCAGCGGAAAACTTTTACGTCAACTCAGTTTGCCTGTTTTACAGGTTGCGGCGATAACTCCCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGCACGTCAACATCATAAAGATGGCAATACGTAACTT
++
+$%'()+-552),54=(&$($((&&#$##&$'*-153./579958713%$%%%$%++2074@HBB674:;D889;BDC>FD=84:3(2(5/>BB0)B)(4?;*-781/-&'$###*+3++)1>B(&&))&,,-.-(00&2?=;>HE@C:/1.&(9E@70%%--%%%%&%./8;:993&.')4544)%(')*8?B?@F=898<LKKD:12831/99($"$##$).01)*0))'6;=;3+0&%).:KLG.'>&//;1$.98:1..,*==><6B87:8<<A55..%&&',**6&'8>:<++)+,,))%,,,+%$##&832::>1@ED.1>?@>EC3'%%$&1.-)&&',+,---**:2+(0+,%&#'()*-',002>=A/0<:>2%%-)*22269<B7%$8665==D62430,$&;;>?C??.<=<>>A<55+&''
+@80de96c0-f65f-45b6-9962-0646f207ffb2
+GATCACTTCGTTCCGGTTACGTAACACTACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTCTAAGCAGACTTTAAAATCGCAACATCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAATAATTACTGTCGTTGACGGACTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAACACATCTAGGTTTTGTCGGGTGGCAATGAAAGGTAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGAAAACACGTCAACTCAGTTTGCCTGTTTACGGTTCGCGACGTACTCGTAAATGCGGTAACTGGGAACTCGTGGAGGGTCTTATCAGAGGCACGTCAACATCTCTAAAGATGAGCAATACGTAATGCTTGA
++
+$#'%#$/05<C?$=%(--&(&%%$$%*.*22;3459/+>>=??=:BA;IA2---'$6''()''-&)%-.%&%%1/776AA:4.2$"$%%%-:,.1:@@A>=9900&&*09=50/6:<8>744;>;7863510/3+068481;>?AE8GG=7'&,..(),378899@?;=859ACCB;7334=>9,#%,@@=?TQMFD@CD==:G:=88<=;221?4/<9)&.(:79&%0;<RSF3+B02**.(%%%&&(112&%%(-110,*32==/:-002(()+5C@<7.**&)4/0,&%&)$1;9>:;'=6GA3DGC<A@,$$%)/B<<996,'$$&-8($%%$&&%$)/62-,-+($$30>@<:8((+,-&%'(+8)*)*-''((153+-,+.,)(./588''&7;=:>>=3//+%$"##$#
+@197e8890-1cb3-4c52-a783-5ef3ce80953a
+CTGCTTCGTTCAGTTACGTATTGCACAAACCAATAACTTTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCGACTGCATGCTTAGTGCACTCACGCGGTATAATTAATAACTAATTACTGTCGTTGACAGGACACGGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAACATCTAGGTTTGTCAGGTGTGACCGAAAGGTAAAGATAGAGCCTTGTCCCTGGTTTCAACGAGAAAACACACGATCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGATATCATTACGTGGCATTGGAGACTCCGTGGAGGAGGTCTTATCAGAGGCGTCAACATCTTAAAGATGGTAATACGTAACTTG
++
+#$$""&+0.<C;BEE<>?<<;9'%$$##&$$555:2''+99<?@@=8C7EAB;+(4<=AGHGCHD:KGH5694''@FOF93;=66-=0==<=:470+1:>=?9636@A=E586?;<=CFJA@8''*CD?A?<<:68@5:?F526AAHFAC-;<?7..C54922//79695=;;:;@??KB=6(''0311106;>ACEHDCA9=B@@EDA>A<=AB=@664/68:363/8,,26<GH-'7$37>7339DI@:CA8?9EA>;)$$$4*45237>B@-?79BEFCG<5892%*+)$%'$%4$%::7;;@/?AE;@CDF@EFH=;;<3><69769444)*((,''+253>63119<=<;8??BJG@C@>4%%-9><=G:=77),$%1'*7:<5640)++58463..',-+(.53341##$$
+@74fdf93b-4411-4aba-8346-7c443da2eead
+GTTGTACTTCGTTCAGTTACGTGACAACACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTCGTGCAGCTGTCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAATAATTACTGTCGTTGACAGGACTGAGTAACTCGTCTATCTTCTGCAAGGCATACGGTTTTCGTCCGTGTTGGCAATCCATCATCGGTTTTTTGTCGGGTGTGACCGAAAGGTAGATGGAGAGCCTTGTCCCTACTGGTTTCTTGAAAACACACGTCACTCGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTATTGGCTTTGGAGGCTGTGGAGGATATATCAGAGGCGTCAACATCTTAAAGATGAGCAATACGTAGCCT
++
+#,-.00//9CDH?<16)*%%&#&#&#$#%(''-4457227??:;=7>;63;:::0,1,.02:;84A>5889<CGGDNE@;555(&$$'#"$/13-%4%%<;59<>>>@<1&(()57355>BA7>/&&><DEMGBC<97</).1-4>=BA;>.>;/,&&:6/+)+(%/1/3470'9-C?C>;*('&&&'&&#&&*%'&,/.*-))*%'%(,%$*%)##''(%$*0%(,139:<0)405C>?BD@BA995,1.((+23311,08,-+/2435/&&%('*%&,&$$&--/,,''$'+).%$'&'(7C/;:;:CBA+%&*/9CE>4801')+**./**%%'1399<635/-$$&%-//&&*%$$''2738018*->A<.:;:75534334556979589:3-*%#$#
+@8fbace70-be6b-4be2-b03c-9265c1693576
+ATATCGCCGTTCAGTTACGTGTACACACCAACCAGCACCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTCCCAAATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCCTCACGCGGTATAATTAATAATAATTACTGTCGTTGACAGGACACGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCGGCCGATCATCAACACACATCTAGGTTTTGTCCGGGTGCTGACCCAGGTAAGATGGAAGCCTTGTCCCACCTAGTTTCAACGAGAAAACACACGTCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTGGAGACTCCAGTGGAGGTCTTATCAGGAGGCACGTCAACATCACTAAAGATGAGCAATACATGTG
++
+##$%%&'*-/,<()13+00'&*'%$$$'(),562%%%%,00660,$**)../0;,.>?ACB3576///2))+&'&'+-+@,015(),@;5<D<;B?,07:688:/&..%-/,$%);00/$$&42/0/.876;<37@:62;@A9EG9A?8,$$591./4,%-999=9@==?=<<?-7494,+,/-/)#&'+31?DJFC>;<@@<94/&&&(*+*9==73;5*00'3&96@1.0;BLSN>%%@'(5;9$@JCBB&&'=@DDDJDD=;(.568895562-13*%:>EB:;<55467,2-&%##&*493.<;?-:7B;2>?=9><7*%%$9:@35657232;5>B@05187***',(-4:669:5')458.-??EGD467(+'(&-++-.7?:,::93%#0/0,04+++6=>999(%%$%#
+@2c03f5fc-9a7b-479a-8d6c-a5bb76a79ed2
+AGTGTGCTTCGTTCAGTTACGTATTGCTACCAACCAATAAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTGATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCACATGGTAGTATAATTAATAACTAGTACTGTCGTTGACAGGACACGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAGCTTTATCTAGGTTTTGTCCGGGTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTGTTTTCAACGAGAAAACACGTCAACTCAGTTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGCACGTCAACATCTTAAAGATAGGCAATACGTAACTGA
++
+)&%&+,6==>:I7D+@<?BA4@@@>/(9;;18812;0,.-(@HH?BEB?@DBC?0(((=;<DHJKDKA.IFD22341131#&-.-09575911+7;4<@?9=B@@@C<9./<6AGCBCDFA3&$$&(2-*033@BBBA;8D:578$E?@EDAB?KDB4,,DC88785'1:E@>B?>?AA@BFPI>A53347<;1))9<@<B@QNG:89DAFCJB?77C>?DACF:5;.-((+061B7DB<AKHD>&&?9>D?CFGJADFDF=EEOG;<7787?7B=>:?>=>AC:4><EB;FC==G7'*;4,-.'&)&(89=6<6BKIDHJB@AA>761..86/<.9;;:-9:@C>$6,23<4AEFGG@?F>@>FEC>C@IF)+079>@?A1+:0,',***78@B1::53(+<<<7*$$$;=CDBA=000,)*$$
+@78cf30ba-8f84-45cd-b9b6-0bc6f3500787
+AAACGTACTTCGTTCAGTTACGTATTGCTAACCAACCAACTTTCGATCTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCCGCAGTATAATTAATAACTAATTACTGTCGTTGACAGGACACGAGTAATATTATCTTCTTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAGCACATCTAGGTTTGTGATTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTAGTTTCAACGAGAAAACACACGTCAACTCAGTTTGCCTCTTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGGCACGTCAACATCTTAAAGATGAGCAATACGTAACTGA
++
+(($%%$#$*/39A9C4:=:''$''&'27,53-/%((,-,$$-3732/12$*120*((6@BFFB*MIE1113667+6;@@;;;-$,6077831;13;7)'%+*,-5&#&--:1+/**9;/*435BDDFA=>@@AG;4:>83:>>C@<C=JG9;99GG<8221.**)&')&*7??=90+''(()8/165AA><--8<8%,4496?866>;;A@:<9:<5/<<$8@(;4>1/.8,>,$%$%)+-=><?@:+*&%;9>?C??C>8DF=>?=:226&%$*0026<3476,718:66/,*.&5>;A=<1:38-&''+&'*==89878CFE7//3-4*56>?9*5598..0101(%(*.;9;9410.0*%$2<?@@??97<47,+>AB:;:CC@>ABFF?D<;;8001ACC5665.))($$##
--- /dev/null	Thu Jan 01 00:00:00 1970 +0000
+++ b/test-data/test.fastq	Fri Aug 27 20:34:32 2021 +0000
@@ -0,0 +1,40 @@
+@8db09f2a-de97-4ebd-80ac-fdd69d356cad
+TGGTCCATGTAACTACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTTAAAATCTGTGTGGCTGTCCTCCGGCTGCATGCTTAGTGCCACTCACGCAGTATAATTAATATTAATTACTGTCGTTGACAGGACCTTTTTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTTTGTCGATCATCAACATCTAGGTTTTGTCCGGGTGTGACCGAAAGGTAGATGGAGAGCCTTGTCCCTGGA
++
+%&%$&$$##$$((%'(:7=@8,>@@C=DB;;>EG;:44456<7CC=<4:)21'-,,06::93;?@?9:=5:3<>?@B?+,0053%(&$$)566)*-)7<9//(&.=0.))*)44;CA@>94--%%21-.77;?=@3*19-((?6$/0/1322(#3572:;??<?ADDFJI?>??@AACFB@LGCADNBQK=?;<@>A@B<.,0%5676830370128=75A<COI;=++A6,-/.D@=DGB>?C;942(+--.-,*423/<=<>=42))&#
+@9518353b-8dcc-460f-9801-2be35ffc6a19
+AAGCATGCTTCGTTCAGTTACGTGGTAACACCAACCAACTTTCGATCTCTTGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTCTCTCGGCTGCAATAGTGCACTCACGTGTAATTAATAACTAATTACTGTCGTTTGACAGGACCTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCCATCTAGGTTTTGTCGGGTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGGAAAACACGTCCACTCAGTTTGCCTGTTTTACAGGTTCGCGACGATGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGGCACGTCAGCATCTTAAAGATGGTAATACGTAACTTAAGCATGCTTCGTTCAGTTACGTGGTAACACCAACCAACTTTCGATCTCTTGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTCTCTCGGCTGCAATAGTGCACTCACGTGTAATTAATAACTAATTACTGTCGTTTGACAGGACCTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCCATCTAGGTTTTGTCGGGTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGGAAAACACGTCCACTCAGTTTGCCTGTTTTACAGGTTCGCGACGATGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGGCACGTCAGCATCTTAAAGATGGTAATACGTAACTTAAGCATGCTTCGTTCAGTTACGTGGTAACACCAACCAACTTTCGATCTCTTGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTCTCTCGGCTGCAATAGTGCACTCACGTGTAATTAATAACTAATTACTGTCGTTTGACAGGACCTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCCATCTAGGTTTTGTCGGGTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGGAAAACACGTCCACTCAGTTTGCCTGTTTTACAGGTTCGCGACGATGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGGCACGTCAGCATCTTAAAGATGGTAATACGTAACTTAAGCATGCTTCGTTCAGTTACGTGGTAACACCAACCAACTTTCGATCTCTTGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTCTCTCGGCTGCAATAGTGCACTCACGTGTAATTAATAACTAATTACTGTCGTTTGACAGGACCTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCCATCTAGGTTTTGTCGGGTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGGAAAACACGTCCACTCAGTTTGCCTGTTTTACAGGTTCGCGACGATGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGGCACGTCAGCATCTTAAAGATGGTAATACGTAACTT
++
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
+@9bd6c9e7-e630-471b-88f5-f0f46e5b3767
+CATCGGTGTTGGCCCGTTTCAGTTACGTAACCTACCAACCAGCTTTCGATCTCTTGTAGATCTATTCTCTAAACGAACTTTAAAATCTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAACTAGTACTGTCGTCTGACAGGACGAAATAACTCGTCTATCTTCTGCAGGCTGTTACGGTTTTCGTCCGTGTACGGCCGATCATCAACACATCTAGGTTTGTCCGGGTGTGACGAAAATGAATGGAAGCCTTGTCCCCTGGTTTCAACGAGAAAACACGTCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCATGCGTGGCTTTGGAACTCCGTGGAGGTCTTATCAGGGCATCAGCATGTAAAGATGAGCAATACGTAACT
++
+'%*+%%&#$%%%((*,88):*-++*,'$$#$%$#++13/*(%$$&/7%'13)034)--2/,7'$798<@8:<D568;@BDCLG216A88#)-;6..1%##''))++$$--)*+&(.,,.>>A2/0//1124664;67:>6014*8315---&'%%%%%%.+$%%)*%'''(*.%''37A8I<87*.,/552)$&%%$&=AB>9200402)&%'&)454?;>--8+'),)/8B/0C:=HB1((>&+030DE,6'01**))'+&&'''.0,020//4.&0(*7?A0/.023(2.42-+.(%=1.;:A3'A:)976;.LLP*%(,%2@=61.0&,,4/?D=&&&)(...$$,7)-,'<<B57///+,4888@344&(&'$$.?2*./(%)//01<''%5;;:<<=6231%%
+@2912dba0-4acb-4b30-aa79-115c7c6e3c6c
+CGATGTACTTCGTTCAGTTACGTAACACTACCAACCAACTTTCGATCTACCCGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCCGGCTGCATCGCCCAGTGCACTCACCAAGTATAATTAATAACTAATTACTGTCGTTGACAGGTTTTCGAGTAACTCTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTTACGACCGATCATCAACATCTATGATGGTTTTGTCAGGTGTGACCGAAAGGTAAGATGGAGAGCCTTGTCCCCCTGGTTTCAGCGGAAAACTTTTACGTCAACTCAGTTTGCCTGTTTTACAGGTTGCGGCGATAACTCCCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGCACGTCAACATCATAAAGATGGCAATACGTAACTT
++
+$%'()+-552),54=(&$($((&&#$##&$'*-153./579958713%$%%%$%++2074@HBB674:;D889;BDC>FD=84:3(2(5/>BB0)B)(4?;*-781/-&'$###*+3++)1>B(&&))&,,-.-(00&2?=;>HE@C:/1.&(9E@70%%--%%%%&%./8;:993&.')4544)%(')*8?B?@F=898<LKKD:12831/99($"$##$).01)*0))'6;=;3+0&%).:KLG.'>&//;1$.98:1..,*==><6B87:8<<A55..%&&',**6&'8>:<++)+,,))%,,,+%$##&832::>1@ED.1>?@>EC3'%%$&1.-)&&',+,---**:2+(0+,%&#'()*-',002>=A/0<:>2%%-)*22269<B7%$8665==D62430,$&;;>?C??.<=<>>A<55+&''
+@80de96c0-f65f-45b6-9962-0646f207ffb2
+GATCACTTCGTTCCGGTTACGTAACACTACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTCTAAGCAGACTTTAAAATCGCAACATCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAATAATTACTGTCGTTGACGGACTGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAACACATCTAGGTTTTGTCGGGTGGCAATGAAAGGTAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGAAAACACGTCAACTCAGTTTGCCTGTTTACGGTTCGCGACGTACTCGTAAATGCGGTAACTGGGAACTCGTGGAGGGTCTTATCAGAGGCACGTCAACATCTCTAAAGATGAGCAATACGTAATGCTTGA
++
+$#'%#$/05<C?$=%(--&(&%%$$%*.*22;3459/+>>=??=:BA;IA2---'$6''()''-&)%-.%&%%1/776AA:4.2$"$%%%-:,.1:@@A>=9900&&*09=50/6:<8>744;>;7863510/3+068481;>?AE8GG=7'&,..(),378899@?;=859ACCB;7334=>9,#%,@@=?TQMFD@CD==:G:=88<=;221?4/<9)&.(:79&%0;<RSF3+B02**.(%%%&&(112&%%(-110,*32==/:-002(()+5C@<7.**&)4/0,&%&)$1;9>:;'=6GA3DGC<A@,$$%)/B<<996,'$$&-8($%%$&&%$)/62-,-+($$30>@<:8((+,-&%'(+8)*)*-''((153+-,+.,)(./588''&7;=:>>=3//+%$"##$#
+@197e8890-1cb3-4c52-a783-5ef3ce80953a
+CTGCTTCGTTCAGTTACGTATTGCACAAACCAATAACTTTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCGACTGCATGCTTAGTGCACTCACGCGGTATAATTAATAACTAATTACTGTCGTTGACAGGACACGGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAACATCTAGGTTTGTCAGGTGTGACCGAAAGGTAAAGATAGAGCCTTGTCCCTGGTTTCAACGAGAAAACACACGATCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGATATCATTACGTGGCATTGGAGACTCCGTGGAGGAGGTCTTATCAGAGGCGTCAACATCTTAAAGATGGTAATACGTAACTTG
++
+#$$""&+0.<C;BEE<>?<<;9'%$$##&$$555:2''+99<?@@=8C7EAB;+(4<=AGHGCHD:KGH5694''@FOF93;=66-=0==<=:470+1:>=?9636@A=E586?;<=CFJA@8''*CD?A?<<:68@5:?F526AAHFAC-;<?7..C54922//79695=;;:;@??KB=6(''0311106;>ACEHDCA9=B@@EDA>A<=AB=@664/68:363/8,,26<GH-'7$37>7339DI@:CA8?9EA>;)$$$4*45237>B@-?79BEFCG<5892%*+)$%'$%4$%::7;;@/?AE;@CDF@EFH=;;<3><69769444)*((,''+253>63119<=<;8??BJG@C@>4%%-9><=G:=77),$%1'*7:<5640)++58463..',-+(.53341##$$
+@74fdf93b-4411-4aba-8346-7c443da2eead
+GTTGTACTTCGTTCAGTTACGTGACAACACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTCGTGCAGCTGTCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAATAATTACTGTCGTTGACAGGACTGAGTAACTCGTCTATCTTCTGCAAGGCATACGGTTTTCGTCCGTGTTGGCAATCCATCATCGGTTTTTTGTCGGGTGTGACCGAAAGGTAGATGGAGAGCCTTGTCCCTACTGGTTTCTTGAAAACACACGTCACTCGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTATTGGCTTTGGAGGCTGTGGAGGATATATCAGAGGCGTCAACATCTTAAAGATGAGCAATACGTAGCCT
++
+#,-.00//9CDH?<16)*%%&#&#&#$#%(''-4457227??:;=7>;63;:::0,1,.02:;84A>5889<CGGDNE@;555(&$$'#"$/13-%4%%<;59<>>>@<1&(()57355>BA7>/&&><DEMGBC<97</).1-4>=BA;>.>;/,&&:6/+)+(%/1/3470'9-C?C>;*('&&&'&&#&&*%'&,/.*-))*%'%(,%$*%)##''(%$*0%(,139:<0)405C>?BD@BA995,1.((+23311,08,-+/2435/&&%('*%&,&$$&--/,,''$'+).%$'&'(7C/;:;:CBA+%&*/9CE>4801')+**./**%%'1399<635/-$$&%-//&&*%$$''2738018*->A<.:;:75534334556979589:3-*%#$#
+@8fbace70-be6b-4be2-b03c-9265c1693576
+ATATCGCCGTTCAGTTACGTGTACACACCAACCAGCACCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTCCCAAATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCCTCACGCGGTATAATTAATAATAATTACTGTCGTTGACAGGACACGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCGGCCGATCATCAACACACATCTAGGTTTTGTCCGGGTGCTGACCCAGGTAAGATGGAAGCCTTGTCCCACCTAGTTTCAACGAGAAAACACACGTCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTGGAGACTCCAGTGGAGGTCTTATCAGGAGGCACGTCAACATCACTAAAGATGAGCAATACATGTG
++
+##$%%&'*-/,<()13+00'&*'%$$$'(),562%%%%,00660,$**)../0;,.>?ACB3576///2))+&'&'+-+@,015(),@;5<D<;B?,07:688:/&..%-/,$%);00/$$&42/0/.876;<37@:62;@A9EG9A?8,$$591./4,%-999=9@==?=<<?-7494,+,/-/)#&'+31?DJFC>;<@@<94/&&&(*+*9==73;5*00'3&96@1.0;BLSN>%%@'(5;9$@JCBB&&'=@DDDJDD=;(.568895562-13*%:>EB:;<55467,2-&%##&*493.<;?-:7B;2>?=9><7*%%$9:@35657232;5>B@05187***',(-4:669:5')458.-??EGD467(+'(&-++-.7?:,::93%#0/0,04+++6=>999(%%$%#
+@2c03f5fc-9a7b-479a-8d6c-a5bb76a79ed2
+AGTGTGCTTCGTTCAGTTACGTATTGCTACCAACCAATAAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTGATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCACATGGTAGTATAATTAATAACTAGTACTGTCGTTGACAGGACACGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAGCTTTATCTAGGTTTTGTCCGGGTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTGTTTTCAACGAGAAAACACGTCAACTCAGTTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGCACGTCAACATCTTAAAGATAGGCAATACGTAACTGA
++
+)&%&+,6==>:I7D+@<?BA4@@@>/(9;;18812;0,.-(@HH?BEB?@DBC?0(((=;<DHJKDKA.IFD22341131#&-.-09575911+7;4<@?9=B@@@C<9./<6AGCBCDFA3&$$&(2-*033@BBBA;8D:578$E?@EDAB?KDB4,,DC88785'1:E@>B?>?AA@BFPI>A53347<;1))9<@<B@QNG:89DAFCJB?77C>?DACF:5;.-((+061B7DB<AKHD>&&?9>D?CFGJADFDF=EEOG;<7787?7B=>:?>=>AC:4><EB;FC==G7'*;4,-.'&)&(89=6<6BKIDHJB@AA>761..86/<.9;;:-9:@C>$6,23<4AEFGG@?F>@>FEC>C@IF)+079>@?A1+:0,',***78@B1::53(+<<<7*$$$;=CDBA=000,)*$$
+@78cf30ba-8f84-45cd-b9b6-0bc6f3500787
+AAACGTACTTCGTTCAGTTACGTATTGCTAACCAACCAACTTTCGATCTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCCGCAGTATAATTAATAACTAATTACTGTCGTTGACAGGACACGAGTAATATTATCTTCTTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAGCACATCTAGGTTTGTGATTGTGACCGAAGGTAAGATGGAGAGCCTTGTCCCTAGTTTCAACGAGAAAACACACGTCAACTCAGTTTGCCTCTTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGGCACGTCAACATCTTAAAGATGAGCAATACGTAACTGA
++
+(($%%$#$*/39A9C4:=:''$''&'27,53-/%((,-,$$-3732/12$*120*((6@BFFB*MIE1113667+6;@@;;;-$,6077831;13;7)'%+*,-5&#&--:1+/**9;/*435BDDFA=>@@AG;4:>83:>>C@<C=JG9;99GG<8221.**)&')&*7??=90+''(()8/165AA><--8<8%,4496?866>;;A@:<9:<5/<<$8@(;4>1/.8,>,$%$%)+-=><?@:+*&%;9>?C??C>8DF=>?=:226&%$*0026<3476,718:66/,*.&5>;A=<1:38-&''+&'*==89878CFE7//3-4*56>?9*5598..0101(%(*.;9;9410.0*%$2<?@@??97<47,+>AB:;:CC@>ABFF?D<;;8001ACC5665.))($$##