Update ingest to accommodate oropouche segments

ingest/Snakefile

@@ -8,6 +8,11 @@ workdir: workflow.current_basedir
 # Use default configuration values. Override with Snakemake's --configfile/--config options.
 configfile: "defaults/config.yaml"
 
+segments = ['L', 'M', 'S']
+
+wildcard_constraints:
+    segment = "|".join(segments)
+
 # This is the default rule that Snakemake will run when there are no specified targets.
 # The default output of the ingest workflow is usually the curated metadata and sequences.
 # Nextstrain-maintained ingest workflows will produce metadata files with the
@@ -17,8 +22,9 @@ configfile: "defaults/config.yaml"
 # TODO: Add link to centralized docs on standard Nextstrain metadata fields
 rule all:
     input:
-        "results/sequences.fasta",
-        "results/metadata.tsv",
+        sequences=expand("results/{segment}/sequences.fasta", segment=segments),
+        metadata=expand("results/{segment}/metadata.tsv", segment=segments),
+        metadata_all="results/all/metadata.tsv",
 
 
 # Note that only PATHOGEN-level customizations should be added to these
@@ -28,6 +34,7 @@ rule all:
 # by build-specific rules.
 include: "rules/fetch_from_ncbi.smk"
 include: "rules/curate.smk"
+include: "rules/nextclade.smk"
 
 
 # We are pushing to standardize ingest workflows with Nextclade runs to include
@@ -41,21 +48,15 @@ include: "rules/curate.smk"
 # to the final metadata TSV.
 # To run nextclade.smk rules, include the `defaults/nextclade_config.yaml`
 # config file with `nextstrain build ingest --configfile defaults/nextclade_config.yaml`.
-if "nextclade" in config:
-
-    include: "rules/nextclade.smk"
-
-else:
-
-    rule create_final_metadata:
-        input:
-            metadata="data/subset_metadata.tsv"
-        output:
-            metadata="results/metadata.tsv"
-        shell:
-            """
-            mv {input.metadata} {output.metadata}
-            """
+rule create_final_metadata:
+    input:
+        metadata="data/subset_metadata.tsv"
+    output:
+        metadata="results/all/metadata.tsv"
+    shell:
+        """
+        cp {input.metadata} {output.metadata}
+        """
 
 # Allow users to import custom rules provided via the config.
 # This allows users to run custom rules that can extend or override the workflow.

ingest/defaults/config.yaml

@@ -119,3 +119,7 @@ curate:
     "abbr_authors",
     "institution",
   ]
+
+nextclade:
+  segment_reference: "../shared/oropouche_{segment}.fasta"
+  min_seed_cover: 0.01

ingest/rules/curate.smk

@@ -8,7 +8,7 @@ REQUIRED INPUTS:
 OUTPUTS:
 
     metadata    = data/subset_metadata.tsv
-    sequences   = results/sequences.fasta
+    sequences   = results/all/sequences.fasta
 
 """
 
@@ -62,7 +62,7 @@ rule curate:
         annotations=config["curate"]["annotations"],
     output:
         metadata="data/all_metadata.tsv",
-        sequences="results/sequences.fasta",
+        sequences="results/all/sequences.fasta",
     log:
         "logs/curate.txt",
     benchmark:
@@ -121,11 +121,11 @@ rule subset_metadata:
     input:
         metadata="data/all_metadata.tsv",
     output:
-        subset_metadata="data/subset_metadata.tsv",
+        metadata="data/subset_metadata.tsv",
     params:
         metadata_fields=",".join(config["curate"]["metadata_columns"]),
     shell:
         """
         tsv-select -H -f {params.metadata_fields} \
-            {input.metadata} > {output.subset_metadata}
+            {input.metadata} > {output.metadata}
         """

ingest/rules/nextclade.smk

@@ -1,95 +1,55 @@
 """
 This part of the workflow handles running Nextclade on the curated metadata
-and sequences.
+and sequences to split the sequences into L, M, and S segments.
 
 REQUIRED INPUTS:
 
-    metadata    = data/subset_metadata.tsv
-    sequences   = results/sequences.fasta
+    metadata     = data/subset_metadata.tsv
+    all_metadata = results/all/metadata.tsv
+    sequences    = results/all/sequences.fasta
 
 OUTPUTS:
 
-    metadata        = results/metadata.tsv
-    nextclade       = results/nextclade.tsv
-    alignment       = results/alignment.fasta
-    translations    = results/translations.zip
+    metadata        = results/{segment}/metadata.tsv
+    sequences       = results/{segment}/sequences.fasta
 
 See Nextclade docs for more details on usage, inputs, and outputs if you would
 like to customize the rules:
 https://docs.nextstrain.org/projects/nextclade/page/user/nextclade-cli.html
 """
-DATASET_NAME = config["nextclade"]["dataset_name"]
 
-
-rule get_nextclade_dataset:
-    """Download Nextclade dataset"""
-    output:
-        dataset=f"data/nextclade_data/{DATASET_NAME}.zip",
-    params:
-        dataset_name=DATASET_NAME
-    shell:
-        """
-        nextclade3 dataset get \
-            --name={params.dataset_name:q} \
-            --output-zip={output.dataset} \
-            --verbose
-        """
-
-
-rule run_nextclade:
+rule run_nextclade_to_identify_segment:
     input:
-        dataset=f"data/nextclade_data/{DATASET_NAME}.zip",
-        sequences="results/sequences.fasta",
+        metadata = "data/subset_metadata.tsv",
+        sequences = "results/all/sequences.fasta",
+        segment_reference = config["nextclade"]["segment_reference"],
     output:
-        nextclade="results/nextclade.tsv",
-        alignment="results/alignment.fasta",
-        translations="results/translations.zip",
+        sequences = "results/{segment}/sequences.fasta",
     params:
-        # The lambda is used to deactivate automatic wildcard expansion.
-        # https://github.com/snakemake/snakemake/blob/384d0066c512b0429719085f2cf886fdb97fd80a/snakemake/rules.py#L997-L1000
-        translations=lambda w: "results/translations/{cds}.fasta",
+        min_seed_cover = config["nextclade"]["min_seed_cover"],
     shell:
         """
-        nextclade3 run \
-            {input.sequences} \
-            --input-dataset {input.dataset} \
-            --output-tsv {output.nextclade} \
-            --output-fasta {output.alignment} \
-            --output-translations {params.translations}
-
-        zip -rj {output.translations} results/translations
+        nextclade run \
+            --input-ref {input.segment_reference} \
+            --output-fasta {output.sequences} \
+            --min-seed-cover {params.min_seed_cover} \
+            --silent \
+            {input.sequences}
         """
 
-
-rule join_metadata_and_nextclade:
+rule subset_metadata_by_segment:
     input:
-        nextclade="results/nextclade.tsv",
-        metadata="data/subset_metadata.tsv",
-        nextclade_field_map=config["nextclade"]["field_map"],
+        metadata = "results/all/metadata.tsv",
+        sequences = "results/{segment}/sequences.fasta",
     output:
-        metadata="results/metadata.tsv",
+        metadata = "results/{segment}/metadata.tsv",
     params:
-        metadata_id_field=config["curate"]["output_id_field"],
-        nextclade_id_field=config["nextclade"]["id_field"],
+        strain_id_field = config["curate"]["output_id_field"],
     shell:
         """
-        export SUBSET_FIELDS=`grep -v '^#' {input.nextclade_field_map} | awk '{{print $1}}' | tr '\n' ',' | sed 's/,$//g'`
-
-        csvtk -tl cut -f $SUBSET_FIELDS \
-            {input.nextclade} \
-        | csvtk -tl rename2 \
-            -F \
-            -f '*' \
-            -p '(.+)' \
-            -r '{{kv}}' \
-            -k {input.nextclade_field_map} \
-        | tsv-join -H \
-            --filter-file - \
-            --key-fields {params.nextclade_id_field} \
-            --data-fields {params.metadata_id_field} \
-            --append-fields '*' \
-            --write-all ? \
-            {input.metadata} \
-        | tsv-select -H --exclude {params.nextclade_id_field} \
-            > {output.metadata}
-        """
+        augur filter \
+            --sequences {input.sequences} \
+            --metadata {input.metadata} \
+            --metadata-id-columns {params.strain_id_field} \
+            --output-metadata {output.metadata}
+        """

shared/oropouche_L.fasta

@@ -0,0 +1,100 @@
+>NC_005776.1 Oropouche virus segment L, complete genome
+AGTAGTGTGCTCCTATTCCGAAACAAACAAAAACAATCTCAAAATGTCACAACTGTTGCTCAACCAATAT
+CGGAATAGGATATTGCACTGCCGTGAACCTGAGATAGCAAAGGATATATGGCGAGATCTATTAAATGATC
+GACACAATTACTTTTCTCGGGAATTTTGCAGAGCTGCAAATCTTGAGTACAGAAATGATGTTCCTGCTGA
+GGATATTTGTGCTGAAGTTCTTGATGGTTATAAAGCAAGGAAAGTTCGCTTTTGTACACCTGATAATTAC
+TTACTACATGATGGAAAGATGTATATAATAGACTTCAAAGTGTCTGTAGACGACCGATCTTCTAGAATCA
+CAAGGGAGAAATATAATGAGATTTTTGGAGAGGTATTCAATCCAGAAGGTGTAGATTTTGAAATTGTTAT
+TATTAGATTAGATCCTTCAAATATGACGATACATGTGGACTCTCGAGATTTCGTGAATACAATTGGGCCG
+ATTACATTAAACATTAGTATGCAATGGTTTTTTGATATGAAAGACTTCTTGTTCGGGAAATTTCGGGATG
+ATGATAAATTCCATGCTATAATAAGTCAAGGAGAATTCACAATGACATTGCCATGGATTGAAGAAGACAC
+CCCAGAATTGCTTACTCATCCTATATACAATGAATTCATGAGTTCAATGCCAGAGGCAGAACAGGCCCTA
+TTCAAGGAAGCATTGGAATTCAAATCATTTGGGGCAGAAAAATGGAATATCTTTTTGAAGGGGGTGATGT
+CAAAGTATGGTGAATATTATAAAGAATTTACTAAAGGACATGCTCATTCTATATTTCTGACAACAGGGGA
+CTACCCCAAGCCAGACAAAGACCAAATTTCAGCAGGTTGGAGAGAAATGGTAAACAGAGTAAGCTCTGAA
+CGTGACATGTCAAATGACATAAATCAGGAAAAACCAAGCATGCATTTTATATGGGCAAAGAATGATTCAA
+ATAGCAACAATAATATACAAAAGCTAATCAAACTATCTAAATCACTGCAAGCTATGAGCGGGACAGGGAG
+CTATGTAAATGCTTTCAAGTCATTAGGGAGATTAATGGATATATCATCAGATGTTAAAAAATATGAATCA
+TTTTGTGGGAAATTGAAATCTCTGGCAAGGTCTAGTATAAAAAAACTTGACAGGAAAATAGAGCCAATAC
+AAATTGGGACTGCAACTGTCTTATGGGAACAGCAATTTAAACTAGATACAGATGTTATAAAAAGAGAAGA
+CAGAATACATTTAATGAAAGATTATCTTGGGATCGGTAAGCACAAATCATTTTCAAAGAAATTAAACAAC
+GACATAAATACTGATAAGCCTAAAATATTAAATTTCAACAATGATGATATAGTCAGGAAATGCAAAGATA
+AATATAATCAAGTCATACATAACCTATCCCAAATCAATGAATTAGATAAGATTGGAAACTACCTAGAGCA
+CTTTTCAGCTAAAATTAGTGCCTGCAGTGTAGAAATGTGGGATTTTATATATAATACAACCAAAACTAAA
+TACTGGCAATGCATCAATGACTATTCCACCCTAATGAAAAACATGTTAGCTGTCTCTCAATATAATAGAC
+ACAATACGTTTAGAATTGTCTCATGTGCAAACAATAATGTATTTGGTCTAGTAATGCCAAGCTCAGATAT
+AAAGACAAAAAAAGCAACTTTAGTCTATGCAATAATGGCTCTCCATAATGAGGAGGCAGAAATAGCAGAA
+CTTGGCTCACTCTACTCAACTTTTAAGACAGCAACAGGATATATTTCAATATCAAAGGCTTTTAGGCTGG
+ATAAAGAAAGATGCCAACGCATAGTATCCTCTCCAGGCTTGTTCCTCATGACAAGCTGCCTATTATTCAA
+CGGTAACAAGAGTTTAGAATTTGATAAATTACTAGGATTTTCATTTTTTACGTCAATATCAATTACGAAA
+GCTATGCTCTCCCTTACTGAGCCTTCACGTTATATGATCATGAACTCGTTAGCAGTTTCCAGCCATGTAA
+GAGAGTATATATCTGAAAAATTCTCCCCTTATACAAAAACATCATTTTCTGTGGTAATGACAGACTTAAT
+CAAGAAGGGTTGCTATTCAGCATATGAACAGAGAAAAAAAGTACAAATAAGAGACATAAAATTAACAGAT
+TATGATATAACACAAAAGGGAGTGGATTCCAAAAGAGATCTTAAATCTATTTGGTTCCCAGGAAAGGTAA
+ACCTGAAAGAATATTTAAACCAAATTTATCTACCATTTTATTTTAACTCTAAAGGATTACATGAAAAACA
+TCATGTCTTGATAGATTTGGCTAAAACAGTACTAGAAATCGAAAAAGAGCAAAGGGAGTCATTACCTGAG
+CCATGGTCAGAGATACCTGCTAAGCGACTGTCACTTAATGTTTTAATTTACTCATTGCAGGAACTGAATT
+TAGATACTTCAAGACATAATTTTGTAAGAAGCCGGGTGGAAAACGCAAATAATTTCAACAGATCTATAAC
+GACAATATCTACTTTTACCAGCTCAAAATCATGCATTAAGATTGGTGATTTTGAAGAAGAAAAAAGAGAA
+AAACTAAGAATGATACAAAAGAAACTTGCAAAGGATATTTCTAAATTAACCATAGCCAACCCAGCATTCT
+TAGATGAGATCACAAACGAACATGAGATAAGGCATTCAACTTATGAGGACTTAAAACAATCTATCCCAGA
+TTACACAGATTATATGTCTGTGAAAGTTTTTGACAGATTGTACGAGAAGATTACTACCAATGAAATAAAT
+GATAAGGAAACAGTCAAGCTGATTCTAGAGACCATGAAAAAACATAAAATATTTCATTTTGGATTCTTCA
+ATAAAGGACAAAAAACAGCCAAAGATAGAGAAATATTTTTAGGTGAATTTGAAGCAAAAATGTGTCTGTA
+CCTTGTCGAAAGAATAGCTAAAGAGAGGTGCAAATTAAACCCTGAAGAAATGATAAGTGAACCAGGCGAC
+TCGAAACTAAGGGTATTAGAGAAGCAATCAGAAGACGAAATCAGGTATATTAGCAATACAATAAAGACAT
+TAGGGAATGCCATAGAGAACTTGCAATCTGGATCTTTAAATTGGGCAGATATATGCGAAAACAAAGCAAG
+AGGACTTAAGATAGAAATAAATGCTGATATGTCCAAATGGAGTGCCCAAGATGTACTTTTTAAATATTTT
+TGGTTGATAGTGCTTGATCCCATCTTATATCCTGCTGAGAGGAAAAGGATAATTTATTTCCTCTGTAATT
+ATATGCAGAAAAGGCTTATAATGCCCGATGAATTGCTCACTACTATATTGGATCAAAGAGTTCCTTATTC
+AAATGACATAATTGGATTAATGACAAACAATTATAGGTCTAATACAGTAGAAATAAAGCGTAACTGGCTT
+CAAGGCAACTTAAATTATACAAGCAGTTACTTACACAGCTGTAGTATGTCTGTGTACAAAGATATAATAA
+GAGAAGCAGCAATATTATTAGAAGGAGAAGCCCTTGTGAACTCAATGGTACATTCTGATGATAATCAAAC
+ATCTATATGTATGGTGCAGAATAAATTACCAGATGACAATATAATTGAATTTTGCATTAAGATATTCGAG
+AAGATATGCTTAACTTTTGGCAATCAGGCAAATATGAAGAAGACATATCTAACTAACTTCATCAAAGAGT
+TTGTTTCTTTATTTAATATACATGGAGAACCATTTTCTATATATGGGAGATTTCTACTCACAGCAGTAGG
+AGACTGTGCCTATCTAGGGCCTTATGAAGATTTAGCAAGTAGGCTATCTGCAACACAAACTGCTATAAAG
+CATGGTTGCCCACCATCACTTGCATGGGTATCTATCGCTCTAAATCACTGGATAACCCACACTACATATA
+ATATGTTGCCTGGCCAAAATAATGACCCGTTACCATTCTTCCCTACTAACAATAGAAGTGAAATACCAGT
+AGAGATGTGCGGAATACTAGAAAGTGATTTATCAACAATTGCACTAACTGGTTTAGAAGCAGGGAATGTC
+ACGTTTCTAACAAATATAGCAAGGAAGTTATCATCCCCAATCTTACAAAGAGAAAGTATTCAAGATCAAT
+ACAATTCTATAGAAAAGTGGGATCTGAGCAAATTATCACAGATCGACATTCTAAGGCTTAAAATGCTCAG
+GTATATATCTCTTGATAGTTCAGTCACATCTGATGATGGTATGGGGGAGACTAGTGAAATGAGATCTCGA
+TCACTTTTAACACCTCGTAAATTCACAACAAGTGGGTCACTTAATAGGTTGAAATCATATAAAGACTTTC
+AAGATATAATAGCAGATGAGGACAAGACAAACGAACTATTTGAGAATTTCATTAGACACCCAGAGTTACT
+GGTTACAAAAGGCGAAACATTTGAAGAATTTGTTAATACGATATTATTTAGGTACAATTCAAAGAAATTC
+AAAGAATCTTTGTCAATACAAAACCCAGCACAGCTTTTTATTGAGCAAATATTATTTTCCAATAAACCAG
+TAATTGACTACACTAGCATACATGACAAGATTTTTGGATTACAAGACATGCCAGGAATTGAAGAACTAGA
+TACAATTATAGGTCGCAAAACATTTGTTGAGAGTTATGTTCAAATCGTAGATGACTTAAGCAATTTAACA
+TTGGATATAAACGATGTCAAGACTATATTTGCCTTTTGTCTTATGAATGACCCACTACTGATCACATCTG
+CTAACAATATAATAATGTCTGTTAAGGGACATAGTCAAGAAAGAATAGGTCAATCAGCATGCAAAATGCC
+AGAGGTCCGAAGTCTAAAACTCATACATTATTCACCAGCAGTTGTTTTGAGAGCCTATGTGAGAGGGCCA
+ACAAATGTACCGAATGTAGATATAGATGAACTTGCAAGGGATCTATCTCATTTAGAAGACTTCATACAAA
+GTACAAAACTCAGAGAAAATATGAGAGAGAGAATAGAAATAAATGAGAAGCGGCACTTAGGAAGGGATTT
+CAAATTTGAAATCAAAGAACTAACTAGATTTTACCAAGTGTGTTATGATTACATAAAGTCTACAGAACAT
+AAAGTCAAGGTATTCATATTGCCATACAAAGTTTTCACATCAATAGAATTCTGCGGGGCACTGACAGGTA
+ACTTGATAAATGACAAATTATGGTACATAACGCATTATCTGAAAAATATAGTGTCTACTACACATAAGGC
+ACAAATTTCTTCTTCACCTGAATTGGAATTGCAAATTGCTGATGAGGCACTAAGACTAGTAGCACATTTT
+GCTGATACTTTCTTGGCATCAGAATCAAGAATACAATTTCTGAAGAAAATTATTGAAGAATTCACATACA
+AAGGGATACCTGTAAAACATTTATACTCAAAAATAAAGAACTCCAAGTTGAGGGTTAAATTTCTAGGGAT
+TCTTTTATGGTTAGATGATCTAACACAGAATGATCTGGATAAATTTGATGCAGATAAATCAGATGAAAAG
+ATTATATGGAATAACTGGCAAGTGTCAAGAGATATGAATACTGGACCAATAGACTTAATGATAAGCGGTT
+ACTCTAGACAGCTGCGGATCACTGGGGAAGATGACAAATTGATTGCTGCTGAATTGCAGGTTACTAGATT
+GTCAGAAGATTTAATTTATAGACACGGTCAGGCAATGTTGAATAAGCCACACGGCTTAAAGCTTGAAAAA
+ATGCAACCTGTGACTGAGATGTCTAAACGATTACATTATATCGTTTTCCAGCAAAGATCACGGAAACGAT
+ACTTCTATTCTATATTACCCACCCAAGTAATTGAGGACCATAATTCTAGAGTTGAATCATCTAGGCTAAG
+CAGAGATTCAAAATGGGTTCCTGTATGCCCTGTTGCAATATCAAAACTCTACCAACAAGGACGGCCTATA
+CTTTCCAAAGTTAGAAATCTGAATATGCAGACTCATTCGCTTTCCAGAATACAAGTTAATGTAGATGAAT
+ATGCCATCACGAGAAGAGCACATTTTCAGAAAATGCCTTTCTTCGAAGGACCATCAATCCCTTCTGGTGG
+TATGGATTTGTCTGAGTTGATGAAATCTACATCCCTATTAAGCTTGAATTATGATAACATAAAAAATGCA
+TCCTTATTGGACATGTCTAGGGTATTTAAGTGCAATGGCAGTGGAGATGACCAAATGGCTTTCGAATTTC
+TATCGGACGAAATTTTGGAGCAAGATGTAGTTGAAGAAATAGAATGCAACCCTATATTTTCTATTAGTTA
+TACAAAAAGAGGAGAATCCAATATGACTTATAAAAATGCTTTCCACAAAGCCTTAATCTCAGAATGTGAC
+AAATTTGAAGAAGCATTTGACTTCCTCGACATGGGATTTTGCTCGAATGAAAATCTTAGTATTCTGGAGG
+AAATACATTGGATAATCAGTTATTTAAAAACAAATCAATGGTCTACGGAACTAGACAATTGTATTCACAT
+GTGCATGTACAGGAATGGATATGATGCAGAATATCATAAATTTGATATACCCTCTAAATTCCTCAAAGAC
+CCAATAAACCGAACAATAAATTGGACTGAAGTCATTGAATTTATATTATTAATTGAAGATTTCCAAACAA
+AAATTGAGCCATGGTCTAGTATGAAGTCACACTTCTGTTCAAAAGCACACAGTGTAGCACTAGAGTGTAT
+GAAAAATGAGAAAAGATCATTGGCAGAATTTGTAGACAAAAGTAAGAAAACTGGCAAATCCAAATTTGAC
+TTCTAAGGTATACACATGTAAAAGTAGTGTTTGTTTCTAAATAGGAGCACACTACT
+

shared/oropouche_M.fasta

@@ -0,0 +1,65 @@
+>NC_005775.1 Oropouche virus segment M, complete sequence
+AGTAGTGTGCTACCGGCAACAAACAGTGACAATGGCGAATTTAATAATTATTTCAATGGTTCTGGGCGTT
+GCCTATGGGCATCCGCTTTCAACAAGTCAAATTGGTGACCGCTGTTTTGCTGGTGGTAACCTCTTCAAGG
+AGATGAACTTGAGTGTAGGACTTGGCGAAATATGTGTTAAAGATGACATATCTATTGTTAAGAGTACTAC
+TGTTTTTAGCAAGAATAAACCAGCTCTTGAGGCAACTACAAAATTTTATAGATCTTTTATTGTGAAAGAT
+TGGTCTGAGTGCAATCCAGTGCTAGACAAATTTGGAAATTTTATGGTTTTAAGTGTTGATGATAATGGCC
+ATATAATACCTAAAATGTATACATGTAGAGCAGCATGTGATATTAGATTAAACAAAGATAATGCCGAAAT
+AATATTATCGTCAACAAAACTCAACCACTTTGAAATTGTCGGGACAACTTCGACATCAGGCTGGTTCAAG
+AACACAATAACAAACAACTTAGAACATACATGCGAGCATGTAACCGTAAATTGTGGTCAGAAATCAGTAA
+AATTCCATGCATGCTTTAGACAACATAGGGGTTGTATAAGATTCTTTAAAGGAACATATATGCCATACTC
+AATGATTGAAGCAATGTGCGTTAACATAGAGCTAATAATTCTGACATTATATATCTTTGCTGCAATTATA
+TTTGCATTAATTATAACAAAGAGTTATGTGGCATACTTGCTCCTGCCGTTATTTTATCCAGTGACGTGGT
+TTTACGGAAAGGTTTATAAAAAGATTAATTCTTGCCCAAATTGTCTACTTGCATCTCACCCTTTCACCAG
+CTGCCCTAAATTCTGTATATGCGGTTCTAGGTTTAGTTGCACAGAAGCTCTTAAGGTACATAGAATGGGA
+AAGGACTGCTTAGGTTATAAGTCTCTAAGCAAAGCTAGACAGATGTGTAAATCGAAAAGCTGGTCCTTCA
+CAGCAGCTATATTAACTGGATTAATATTGATGGAATTCATCTCACCAATTGCAGGAGAGAGAATGTATAA
+GCTTGAGGAGCTGGCTGATGAGTTTATAAAGCTAACAGAGCAGGTAAATATTTTAGAAAGAGAAATGGAA
+GTCTTGAAGCAGTCAATAATTGTTATGTTTGCAATCATTTTGGTATTGTTGCTTTTTGAAAATATGATAT
+TCAACCGGCTTTTCCGCATTTTTTATCGTTCTTGCTCCATGTGCGGCTTGATACATTATAGACCAGGACT
+CAAGATCGATTTAACCAAAACTAACAGATGTGGTAGTTGCATCTGTGGATTCGACGAGCAGCAATCATCT
+GGTTTTGAATATGAAATATTTCTAAAGGATATGCATGTCCAGAAAGAATCATGCAAATTTAGTCCAAGAG
+TAAACCACTTTAGGAATATTAAAGCTTTATTATTTGCACTTGTAATCTGTGCTACTATATATACAGTTTA
+TGCAGATGAAGATTGCTTATCTAAAGATATTAAAATAACATATCAAGAGTTGCACAATTGCATAGGTCCA
+AAGATAATGGGCGATACTTGTGTATCAAAGAGTGAATTGTATTCTGATTTATTCTCAAAAAATCTAGTCA
+CAGAATATGATAAAAAATATTTTGAACCCGATACTGTAAATGATCAATTCAACAAGATAGAGTTTGCCCA
+AGATGCACATAGAATGATACTATTAGAAAGAATACTTTACAAGACAGAATGTGAAATGCTAAGCTTAAAA
+AAGAACAGTGGACCTTATAATGTTGCCTGGCGCACATATTTAAAAAATCATAATATAGACTTATGCAGTA
+GGCACAACTATAAAATGATATGTCAGTGTATTAATACACACTCTATGTGTAATAATACAGATATAGATTA
+TAACAAAGAGATCGAAACATATTATAAAAGCAATGCTGCAGCCTATAGAGCTGACCTGAACACCATAATG
+GACACATTAAAAACAGCATTCAGAGGGCTTACAAAGGTCCTAATTGAAAATTATATAGAAAAAGATGACT
+CTGATGCTCTTAAGGCCTTATTTAGTAATATTACTGATTCTGTTCAGGACAATTATCAGATGATTGGAAT
+ATTGAAATTTGCCAGTAAGCTATTAGATATTAACCTTGGTAGATCCACCCGCTCAGCTCATCATTCTATA
+ATGACAAACGAAATTCCCAAATCTAATCCTTTTACTGATTACAGTTATTCTAATCTGAATATAAAGGAGT
+GCATGTCACCAGAGAGCTTGAAATGTTTTAAGAAGAGAGGCAGTACACCGCATACAAACCACCTGCTTTG
+CAAGATAGACAACAAATATAAAGCATTTGACTGGCCAGAGATAGAAACAATTCAAAAAGGTCAAAAATTG
+TGCCTTGGTGACAGTCATTGCAACTTGGAATTTACAGCCATAACTGCAGACAAGATTATGTCATTAACTA
+ACTGTTATAAAGAATCTTTCACTGCGCAACCAGCAGATATGCAGGCTGGAATTAAAAAATGCTCAGCTGA
+TGAAATTGGAGAATGTACAACACTTGAAGACAAGACATGGCCAATTATATTCTGTGGTGGCAAATATTAC
+TACTCTGATAGCAAGGAGCATGCCAAGGACGGTTCTATCAACAATTATTGTTTAACCAACAAATGCTCTG
+AACAACGTTTCCCAATACATGAAAATTGGTTCAAAAAATGTAATTGGGACAAGACCCACAAAGAATTTAG
+CACTATGAGACAAATAAACTATAATGATATAACTTCTTATAGAAAGGCAATAGAATCTGAGATAGGAACA
+GATTTGATGACACACCATTATAAGCCCACAAAGAACCTTCCACATGTTGTACCAAGGTATCATAGTATAG
+ATGTACAAGGTACAGAATCAACAGAGGGTATTATCAATGGTTTCATACAAAATACAATACCTGCTATTAG
+TGGTCTAGGTGTCGGTTATCATCTGAATTTTCAAAGCAATCAGTTATTTGACATAGTTATTTTTGTGAAA
+AAAGCTGTATATAAAGCACAGTACCAGAAAGCATACACCACAGGTCCTAGCATATCCATAAATATTGAAC
+ACAACGAAAGATGCACAGGCCACTGCCCCGAAAAGATTCCTGCAAAAGAAGGTTGGTTAACATTTTCTAA
+AGAGCATACTAGTTCTTGGGGATGTGAAGAATATGGCTGTCTTGCTATAGATACAGGCTGTCTATATGGC
+TCTTGTCAAGATGTCATACGCCCAGAATTAGATGTTTATAAGAAGATTGGCAGTGAGGTATCACTGATAG
+AAATATGTATAACTTTGCCTCATGAAACTTATTGCAATGATATGGACATATTAGAACCGATTATAGGAGA
+TAAATTAAGTGCATCCTTTCAGAATACACAGACTAATCAACTTCCGACCCTTATAGCTTATAAAAAAGGG
+AAGATATACACAGGTCAAATCAACGATATTGGAAACACAGCATTGCAGTGCGGCTCAATCCAAGTAATCA
+ATGGATCAACTATCGGAACTGGCAGCCCCAAATTTGATTATATATGCCATGCTATGAGAAGGAAAGATGT
+TATTGTAAGGAAGTGTTTCAATGACAATTATCAGTCGTGCACTAGGCTAGAAAAACGGAACGACTTGATC
+CCTTATAGAAAAGGCGATGTCATCGAAGTATCAAAAACAGGATCGAACATGGGCCAAATGACATTTAAAA
+TAGAACTAGGTGATATCAACTATAAAATATTTACTAAATCAATAGATCTTCAAATGTCTGGCATTTGTGC
+TGGGTGCATTGACTGTGCTGAAGGCATTTCATGCTCTATAAATGCTGAAGTGCCAGCCGAAACAGTCTGC
+CATTGCAAAACAAACTGTGAAGATTTTATAAACAATATAGTGTTTTCACCGCAAATCAAGAATTACAATA
+TAAAAGTCCACTGTAAATCTAAAGTTGAAAAAATAACAGCACATATCTGTGGGAGAGACATTGATTTACA
+GTTAACAATTAAGCCATACAATCAGAAGATAGACTTGTCGCAGTTAGATGAATCAAATTACATAAGGGAA
+GAGGACCTCCAGTGTGGAACATGGTTATGTAAAGTGCAAAAAGAGGGGATTGATATTATATTTAAAGGAT
+TATTTTCAGGCTTAGGGAGATATTGGACCATATTAATTTATTCAATTATAGGTGTGGTAATAATTGTGAT
+ATTGGTTTATATCCTGTTGCCCATAGGTCGGCTTTTAAAAGCATTCCTAATCAGACATGAAATAGAATAT
+GCCATGGAGCAGAAAATCAAGTAGATTTGGCTAAAAAGGGTAGGCAGGTCTAAAATCAGGTATAAATAAA
+ATTCATATAAATAAAGTCAAAAATTGTTGTCGGTAGCACACTACT
+