Setting outgroups for the "Dengue virus DENVx genotypes" dataset #67

Addressing the issue of cross-serotype samples being falsely assigned to genotypes within a specific serotype. For example, when querying all DENV1-DENV4 samples against a DENV4 dataset, there were false-positive "DENV4II" genotype calls for samples that were DENV1-3. The cross-serotype samples should be "unassigned".
nextstrain · Jun 20, 2024 · 84d6de2 · 84d6de2
2 parents 26a536a + d3c39c7
commit 84d6de2
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Showing 14 changed files with 109 additions and 18 deletions.
diff --git a/nextclade/config/clades_genotypes_denv1.tsv b/nextclade/config/clades_genotypes_denv1.tsv
@@ -0,0 +1,15 @@
+clade	gene	site	alt
+DENV1/I	E	461	V
+DENV1/I	E	484	L
+DENV1/I	M	122	R
+DENV1/II	E	345	A
+DENV1/II	E	432	M
+DENV1/III	E	297	V
+DENV1/III	M	118	R
+DENV1/IV	E	339	S
+DENV1/IV	M	72	E
+DENV1/IV	E	88	T
+DENV1/V	NS1	324	R
+DENV1/V	NS2A	142	P
+DENV1/V	NS3	185	K
+DENV1/V	NS5	834	E
diff --git a/nextclade/config/clades_genotypes_denv2.tsv b/nextclade/config/clades_genotypes_denv2.tsv
@@ -0,0 +1,23 @@
+clade	gene	site	alt
+DENV2/AM	E	71	D
+DENV2/AM	E	81	T
+DENV2/AM	E	129	I
+DENV2/AM	NS1	21	V
+DENV2/AM	NS1	73	S
+DENV2/AM	NS1	99	V
+DENV2/AM	NS1	170	R
+DENV2/AA	E	491	A
+DENV2/AA	M	15	G
+DENV2/AA	M	39	I
+DENV2/AI	E	484	I
+DENV2/AI	NS1	222	N
+DENV2/AI	NS5	687	I
+DENV2/AII	NS1	51	Q
+DENV2/AII	NS2A	142	R
+DENV2/AII	NS3	160	S
+DENV2/C	E	71	A
+DENV2/C	E	149	N
+DENV2/C	E	462	V
+DENV2/S	E	59	F
+DENV2/S	E	236	M
+DENV2/S	E	432	V
diff --git a/nextclade/config/clades_genotypes_denv3.tsv b/nextclade/config/clades_genotypes_denv3.tsv
@@ -0,0 +1,9 @@
+clade	gene	site	alt
+DENV3/I	M	128	F
+DENV3/I	E	68	V
+DENV3/II	M	57	A
+DENV3/II	NS5	749	K
+DENV3/III	E	132	Y
+DENV3/III	E	301	T
+DENV3/IV	NS1	139	S
+DENV3/IV	NS5	638	P
diff --git a/nextclade/config/clades_genotypes_denv4.tsv b/nextclade/config/clades_genotypes_denv4.tsv
@@ -0,0 +1,9 @@
+clade	gene	site	alt
+DENV4/I	E	429	L
+DENV4/I	NS1	98	S
+DENV4/II	E	265	A
+DENV4/II	E	46	T
+DENV4/II	NS1	246	S
+DENV4/S	E	132	V
+DENV4/S	E	154	S
+DENV4/S	E	162	T
diff --git a/nextclade/config/config_dengue.yaml b/nextclade/config/config_dengue.yaml
@@ -21,13 +21,19 @@ filter:
     denv3: '36'
     denv4: '36'
 
+outgroup:
+  denv1: "config/outgroup.fasta"
+  denv2: "config/outgroup_denv2.fasta"
+  denv3: "config/outgroup.fasta"
+  denv4: "config/outgroup.fasta"
+
 refine:
   root_id:
-    all: "Reconstructed_root_sequence_of_https_nextstrain_org_dengue/all/genome"
-    denv1: "Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv1/genome"
-    denv2: "Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv2/genome"
-    denv3: "Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv3/genome"
-    denv4: "Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv4/genome"
+    all: ["Reconstructed_root_sequence_of_https_nextstrain_org_dengue/all/genome"]
+    denv1: ['Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv1/genome', 'Reconstructed_root_sequence_of_https_nextstrain_org_dengue/all/genome']
+    denv2: ['Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv2/genome', 'Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv4/genome']
+    denv3: ['Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv3/genome', 'Reconstructed_root_sequence_of_https_nextstrain_org_dengue/all/genome']
+    denv4: ['Reconstructed_root_sequence_of_https_nextstrain_org_dengue/denv4/genome', 'Reconstructed_root_sequence_of_https_nextstrain_org_dengue/all/genome']
 
 traits:
   sampling_bias_correction: '3'
@@ -41,7 +47,7 @@ traits:
 clades:
   clade_definitions:
     all: '../phylogenetic/config/clades_serotypes.tsv'
-    denv1: '../phylogenetic/config/clades_genotypes.tsv'
-    denv2: '../phylogenetic/config/clades_genotypes.tsv'
-    denv3: '../phylogenetic/config/clades_genotypes.tsv'
-    denv4: '../phylogenetic/config/clades_genotypes.tsv'
+    denv1: 'config/clades_genotypes_denv1.tsv'
+    denv2: 'config/clades_genotypes_denv2.tsv'
+    denv3: 'config/clades_genotypes_denv3.tsv'
+    denv4: 'config/clades_genotypes_denv4.tsv'
diff --git a/nextclade/config/outgroup.fasta b/nextclade/config/outgroup.fasta
@@ -0,0 +1,2 @@
+>Reconstructed_root_sequence_of_https_nextstrain_org_dengue/all/genome
+AGTTGTTAGTCTACGTGGACCGACAAGGACAGTTTCGAATCGGAAGCTTGCTTAACGTAGTTCTAACAGTTTTTTTAATTAGAGAGCAGATCTCTGAGATGAACAACCAACGGAAAAAGGCGGGAAAACCGCCTTTCAATATGCTGAAACGCGAGAGAAACCGCGTGTCAACTGTACAACAGTTGGCGAAGAGATTCTCAATAGGATTGCTGTAAGGGCAAGGACCATTAAAATTGGTTATGGCTTTCATAGCATTCCTACGATTCCTAGCCATCCCACCAACAGCAGGGATTTTGAAAAGATGGGGAACATTCAAGAAGTCTAAGGCTATCAAGGTACTGAGAGGATTCAGGAAGGAGATCGGAAGCATGCTGAACATTTTGAACAGGAGAAAAAGAACAGCAATCATTCTCATGATGCTGATTCCAACAGTAATGGCGTTCCATTTGACCACACGAGATGGAGAGCCGCACATGATTGTGAGCAAGCATGAAAGAGGAAAATCCCTTCTGTTTAAGACAGCGGATGGAATCAACATGTGCACCCTCATTGCCATGGATCTGGGTGAGTTGTGTGAGGACACAATCACGTACAAATGCCCCCTTCTCACCGAAACCGAACCAGAAGACATTGACTGCTGGTGCAATCCCACGTCCACATGGGTGACCTATGGGACGTGTACCCAAACTGGAGAACACAGACGAGAGAAAAGATCAGTGGCACTAGCCCCACATGTGGGAATGGGACTGGAAACAAGAACTGAAACATGGATGTCATCAGAAGGAGCTTGGAAACAAGCCCAGAGAGTAGAGACTTGGATTCTGAGACACCCAGGATTCACGATAATGGCAGTATTTCTGGCATACACCATAGGAACAACATATATCCAGAGAGTCGTTATTTTTATCCTACTGATGCTGGTCGCCCCATCCATGACAATGCGATGCGTGGGAGTAGGAAACAGAGACTTTGTGGAAGGAGTTTCAGGAGGAACGTGGGTTGACGTGGTGCTAGAACATGGAAGTTGTGTCACAACCATGGCAAAGAACAAACCAACACTGGACTTTGAACTGATGAAGACAGAAGCCAAACAACCGGCCACACTAAGGAAATTCTGCATTGAAGCTAAAATAACCAACACAACAACAGAATCAAGATGTCCAACACAAGGGGAACCCAATCTAGTAGAGGAGCAGGACCAGAAGTTCGTCTGCAAACAAACGATGGTGGACAGAGGATGGGGCAATGGCTGTGGTTTGTTTGGAAAAGGAGGCATTGTGACATGTGCGAAGTTTACATGCTTGAAGAAAATAGAAGGAAAAGTGGTGCAACCTGAAAACTTAGAATACACCGTGGTCATAACAGTCCACACAGGAGACGAACACGCAGTGGGAAATGACACAACAAAACATGGAATGACGGCCAAGATAACACCCCAGGCTTCCACCGCAGAAGCTAAACTGACAGACTATGGAACTCTCACGCTGGATTGCTCACCAAGAACAGGTCTCGACTTTAATGAGATGGTCTTGTTGAAAATGAAAAACAAAGCATGGCTTGTACACAGACAATGGTTCCTAGACCTACCTCTACCATGGACATCAGGAGCTGACACATAAGAAGCAAATTGGAACCAGAAAGAGATGCTGGTGACATTCAAAAATCCTCATGCAAAGAAACAGGATGTAGTCGTCCTAGGATCCCAAGAAGGAGCCATGCACACAGCACTCACAGGAGCTACAGAAATCCAAACGTCAGGAGGAAACTTAATGTTTGCAGGACACCTGAAGTGCAGACTGAGAATGGACAAACTGAAACTCAAAGGGATGTCATACGCGATGTGCACAGGAAAGTTCAAAATTGAGAAAGAAATGGCAGAAACACAGCATGGAACAATAGTCATTAAGGTTAAGTATGAAGGAGATGGTGCACCATGCAAGATCCCTTTTGAGATAAAGGATGTGAAAAAGAAAAATGTCATTGGGCGACTGATCACAGCCAACCCAATAGTCACAGAGAAAGACAGTCCAGTCAACATTGAAGCAGAACCTCCTTTTGGGGACAGCTACATCGTGATAGGAGTAGGAGAAAGAGCATTGAAACTCAACTGGTTCAAGAAAGGAAGCTCCATTGGGAAAATGTTTGAGGCAACTGCCAGAGGAGCAAAAAGAATGGCCATCTTGGGAGACACAGCCTGGGATTTTGGATCTGTAGGAGGAGTGTTCACATCAGTAGGAAAGGCGGTGCACCAGGTTTTTGGAACTGTCTATGGGGCTATGTTTAGCGGAGTCTCATGGACCATGAAAATCCTAATAGGGGTTCTCATAACATGGATAGGAATGAATTCAAGAAGCACTTCCCTGTCAATGACATGCATAGCAGTAGGAATCGTCACACTGTACCTGGGAGTCATGGTGCAGGCGGACATGGGTTGTGTTGTGAACTGGAAAGGCAAAGAACTGAAATGTGGAAGTGGAATTTTTGTCACTGACAACGTGCACACTTGGACAGAACAATACAAATTCCAACCAGACTCCCCAACAAGACTGGCGTCAGCCATTCCGAAAGCTCATGAGGAGGGAGTCTGTGGAATCCGATCAGCAACAAGACTGGAGAATCTCATGTGGAAGCAAATAACAAATGAATTGAACCACATCCTATGGGAAAATGAAATCAAACTAACTGTAGTGGCAGGAGACATTAAAGGAATCTTGGAGAAAGGGAAAAGAACACTTACACCTCAACCCATTGAGCTGAAATATTCATGGAAAACATGGGGAAAAGCAAAAATACTCACAGCAGAAGTACAAAATAACACCTTCCTCATAGACGGTCCAGACACACCAGAATGCCCTAATGACCAAAGAGCATGGAATTTTCTGGAAGTTGAGGACTATGGATTTGGAATCTTCACGACCAACATATGGCTGAAATTGCGAGAAGAGTACACCGAAGTGTGTGACCACAGGCTAATGTCAGCAGCCATCAAGGACAACAGGGCCGTCCATGCTGACATGGGCTATTGGATAGAAAGCGCAAAGAATGAGACCTGGAAGATAGAGAAAGCATCCTTCATTGAAGTTAAAACCTGCCTCTGGCCAAAATCACACACTCTTTGGAGCAATGGAGTGCTAGAAAGTGAAATGATAATTCCAAAGATTTTTGCTGGACCAATTTCACAGCACAACTACAGACCAGGCTATTACACACAAACAGCAGGACCATGGCACTTAGGCAAATTGGAGATGGACTTTGATTTTTGTGAAGGAACCACAGTTGTAGTGACTGAGGACTGTGGAAATAGAGGCCCATCTTTGAGAACAACCACAGCCTCAGGAAAACTGATAACTGAATGGTGCTGCCGATCTTGCACATTACCTCCCCTAAGATACATAGGAGAGGATGGATGCTGGTATGGGATGGAAATCAGACCCTTGAATGAGAAAGAAGAGAACCTGGTCAAGTCATTGGTCACAGCCGGGCATGGAAAGGTGGACAATTTTTCAATAGGACTCTTGTGCATGGCATTGTTCATGGAAGAGGTGCTGAGAACCAGAGTTGGCAGAAAACACATGATATTAGTAGTAGTGGTTACCTTCGTGACCCTCATCACAGGGAATATGACATGGAGAGACCTGGTAAGAGTACTTATCATGGTGGGAGCCACTATGTCTGATAGAATGGGGGGCGTGACTTACCTAGCTTTAATGGCAGCTTTCAAAATGAGACCAATCTTCGCAGTGGGATTGTTCTTGAGGAAACTGACTTCAAGAGAAATAATACTGATGACAATAGGAATGGCCCTGGTAACATCGTTCCAAATACCAGATGCCATTGTGGAACTGACTGATGGAATAGCATTGGGGATAATGGTGCTTAAAATGGTGACACATTTTGAAAAATACCAGTTGGGAGCGACCATCCTGTCCTTGACGTGCATCAAAACAACTATGACATTGCATAATGCATGGAAAACAATCACCATTATATTGGCAGTAGTTTCTCTTTTCCCATTGTGCCTGACTTCCTCCCTTCAAAAACAGACAGATTGGATACCAATAACATTGACAATCATAGGAGTCAAACCACTACCCATGTTTCTAATAACTTTAACAAAAACCAGCAAAAAAAGAAGTTGGCCTCTCAATGAGGGCATTATGGCTGTTGGAATGGTGAGCATTCTAGCAAGTTCTCTCCTAAAGAATGATGTTCCCATGGCTGGACCATTAGTGGCTGGAGGCTTGCTCATAGCGTGCTACGTGATAACTGGCAGCTCAGCAGACTTGTCACTGGAGAAAGCCGCCGATGTGAAATGGGAAGAAGAGGCAGAGATATCAGGAGCCAGCCCAATCATATTAGTCACAGTAGAAGAAGATGGTACCATGTCAATAAAGGATGAAGAGGAAGAGAACACACTAACCATACTCATAAAAACAGCACTGCTAGTAGTGTCAGGCCTCTTCCCATTGTCAATACCAGTCACAATGGCGGTATGGTACATGTGGCAAATGAAAAAACAAAGATCCGGAGTGCTGTGGGACGTCCCCTCACCCCCAGCAGTGGAAAAAGCTGAACTGGAAGATGGAGTCTATAGAATCAAGCAAAGAGGGATTTTTGGGAAATCCCAAGTTGGAGTTGGAGTATACAAAGAAGGAGTATTCCACACAATGTGGCACGTCACAAGAGGAGCTGTCCTCATGCATAAAGGGAAGAGACTGGAACCATCTTGGGCTGACGTCAAGAAAGACTTGATATCATATGGAGGAGGTTGGAGGCTTGAAGGAAAATGGAAAAAAGGAGAAGAAGTGCAGGTCTTGGCCGTAGAACCTGGGAAAAATCCAAAAAACGTCCAAACAAAACCTGGCCTTTTCAAGACCACTACAGGAGAAATAGGAGCCGTAGCCCTGGACTTCAAACCTGGAACATCAGGATCTCCCATCATTAACAAAAAAGGAAAAGTTGTGGGTCTCTATGGAAATGGAGTGGTTACAAAAAGTGGAGCCTATGTCAGTGCCATAGCGCAAACTGAAAAAATTGAAGAGCCGAATCCAGAGATTGAAGATGACATCTTTAGAAAGAAAAGATTAACCATAATGGACCTACATCCAGGAGCGGGAAAGACAAAAAGATATCTCCCAGCCATAGTCAGAGAGGCCATAAAAAGAAGCCTGAGAACTCTAATCTTGGCTCCCACAAGAGTGGTGGCAGCTGAAATGGAAGAAGCCCTAAGAGGACTCCCAATAAGTTATCAGACCCCAGCCATGAAGGCTGAACACACAGGAAGGGAGATTGTAGACCTAATGTGCCATGCCACTTTCACCATGAGACTGCTGTCACCAATCAGGGTTCCAAATTACAACCTGATAATAATGGATGAAGCACATTTCACAGACCCAGCCAGCATAGCGGCTAGAGGATACATCTCAACCCGAGTGGAAATGGGAGAGGCAGCTGCAATTTTTATGACAGCCACTCCCCCAGGAACGACAGATCCCTTTCCTCAGAGCAACGCCCCAATCAAAGATGAAGAAAGAGAAATTCCTGAACGCTCATGGAATTCAGGATATGAATGGATCACAGATTTTCAAGGGAAGACAGTGTGGTTTGTTCCAAGCATAAAAGCCGGAAATGACATAGCAAACTGCTTGAGAAAAAATGGAAAAAGGGTGATTCAACTGAGTAGGAAAACCTTTGACACAGAATATCCAAAAACAAAAATCAATGACTGGGACTTTGTGGTCACAACAGACATTTCCGAAATGGGAGCCAATTTCAAAGCTGAGAGAGTGATAGACCCAAGGAGATGCCTGAAGCCAGTTATACTGACAGATGGTCCAGAGCGGGTGATTCTGGCAGGACCCATGCCAGTGACCCTAGCAAGTGCTGCCCAAAGAAGAGGGAGAATAGGAAGGAACCCAAAAAAAGAAAATGACCAGTACATTTACATGGGAGAGCCTCTAAAAAATGATGAAGACCATGCTCACTGGACAGAAGCAAAGATGCTCCTTGACAACATCAACACACCAGAAGGGATCATCCCAACCCTGTTTGAACCAGAAAGGGAAAAGACTGAAGCCATAGACGGTGAATACCGCCTGAGAGGGGAAGCAAGGAAAACCTTTGTGGAACTAATGAGAAGAGGAGACCTACCAGTTTGGCTGGCCTACAAAGTAGCTTCAGAAGGCATCAAATACACAGACAGAAGGTGGTGCTTTGATGGAGAAAGGAACAATCAAATTTTGGAGGAGAACATGGAAGTGGAAATCTGGACAAAAGAAGGAGAAAGGAAGAAATTAAGACCTAGATGGTTGGATGCTAGAACTTATTCTGACCCACTGGCACTAAAGGAATTCAAGGAGTTTGCAGCTGGAAGAAAGTCCATAACCCTTGACCTAATAACAGAAATAGGTAGACTGCCAACTTACTTGACCCACAGGGCCAGAAATGCCCTGGACAACTTGGTCATGCTGCACACGACAGAAAAAGGAGGAAGGGCCTACAAACATGCTCTGAATGAACTACCAGAAACCCTGGAAACATTCATGCTCCTAGCTCTGTTGGCTGCGATGACAGGAGGAATCTTTCTGTTCTTGATGTAAGGAAGAGGTATAGGGAAAATGTCCATGGGACTGATCTGCATAATGGCAGCCAGCGGCCTGCTATGGATGGCAGAAATCCAGCCCCATTGGATAGCGGCCTCAATAATACTGGAGTTTTTTCTCATGGTACTGCTTATTCCAGAACCAGAAAAGCAGAGAACTCCCCAGGACAACCAATTGACCTATGTCGTGATAGCCATCCTCACAATGGTTGCAACAATAGCAGCCAATGAAATGGGATTGTTGGAAAAAACAAAGAAAGATCTTGGGTTTGGCCATGTAGCAACTGAAAATACAATTCTGGACGTAGATCTACGTCCAGCATCAGCATGGACACTCTATGCAGTGGCCACAACAATTATCACACCAATGTTGAGACACACCATAGAAAACTCCTCAGCAAATGTGTCCCTGACAGCCATTGCCAACCAGGCAGCGGTCTTGATGGGTCTTGGAAAAGGATGGCCATTATCGAAAATGGACATAGGAGTTCCACTTCTCGCCTTGGGATGCTATTCACAAGTGAACCCAATAACTCTCACAGCAGCACTACTTTTGCTAGTGGCACATTATGCCATAATAGGTCCAGGACTGCAAGCAAAAGCCACTAGAGAAGCTCAGAAAAGGACAGCAGCTGGAATCATGAAGAATCCAACGGTGGATGGGATAACAGTAATAGACCTAGATCCAATACCCTATGATCCAAAATTTGAAAAGCAGTTAGGACAAGTAATGCTCCTGGTCCTTTGTGCAGCACAGCTACTGTTGATGAGAACTACATGGGCTTTGTGTGAAGCCTTAACCCTAGCCACAGGACCAATATCAACACTGTGGGAAGGAAATCCAGGGAAATTCTGGAACACAACGATAGCTGTGTCCATGGCTAACATTTTCAGAGGGAGTTATTTGGCCGGAGCTGGACTTGCTTTTTCTATCATGAAGAATGCAGGAAGCGCAAGGAGAGGAACTGGGACCATAGGAGAAACACTGGGAGAAAAATGGAAAAGACAACTAAACCCATTGGGCAGGAAAGAATTCGAAATATACAAGAAAAGTGGAATTCTAGAAGTGGACAGAACCGAAGCCAAAGAAGGCTTGAAAAGAGGAGAAACAACCCACCATGCCGTGTCGAGAGGCTCCGCAAAACTGAGATGGTTCGTTGAGAGAAACATGGTCAAACCAGAAGGGAAAGTCGTAGACCTTGGTTGTGGAAGAGGAGGCTGGTCATATTACTGTGCGGGACTGAAGAATGTGACAGAAGTGAAAGGATACACAAAAGGAGGACCAGGACATGAAGAACCAATCCCTATGTCTACATATGGATGGAACCTAGTCAAGCTTCAGAGTGGAGTGGATGTGTTCTTCACTCCACCTGAGAAATGTGACACCCTGTTGTGTGACATTGGGGAATCATCACCAAATCCAACAGTAGAAGAAGGAAGAACATTAAGAGTTCTGAAGATGGTGGAACCATGGCTCAACAACAACACTCAATTTTGCATAAAAGTTCTCAACCCCTACATGCCAACAGTCATAGAAAAACTGGAAAAACTACAAAGGAAACATGGAGGAATCCTTGTGAGAAATCCACTCTCACGAAATTCCACCCATGAGATGTACTGGGTGTCCAATGCTTCAGGAAACATTGTGTCTTCAGTGAACATGATATCAAGAATGTTGCTGAACAGATTCACAATGAGACACAGGAAACCCACTTATGAGAAAGACGTGGACCTTGGAGCTGGAACACGAAATGTCGCAATTGAAACAGAAACACCAAACATGGATATCATTGGGAAAAGGATAGAGAAAATAAAAGAAGAGCATAAAACAACATGGCACTATGATCAGGACAATCCATACAAAACATGGGCTTACCATGGAAGCTATGAAGCCAAACCGACAGGATCAGCATCATCCATGGTGAATGGAGTGGTGAGACTGCTGACAAAACCCTGGGACGTGATTCCCATGGTGACACAGATGGCAATGACAGACACAACTCCCTTTGGACAACAGAGAGTTTTTAAAGAGAAAGTGGACACGAGAACACCAAAACCAAAACCAGGCACAAGAAAAGTTATGAAGATCACAGCCGAGTGGCTTTGGAGATTCCTTGGAAGGAAAAAAAAACCCAGAATGTGCACAAGAGAAGAGTTCACAAAAAAAGTTAGAACCAACGCAGCTATGGGCGCCGTGTTCACTGATGAGAACAAATGGAACTCAGCAAGTGAAGCTGTTGAAGACGAACGGTTTTGGGAACTGGTGGACAAAGAAAGGGATCTTCACCAAGAAGGAAAGTGTGAAACGTGTGTCTACAACATGATGGGAAAAAGAGAGAAGAAACTAGGAGAGTTTGGCAAAGCAAAAGGCAGCAGAGCAATATGGTACATGTGGCTGGGAGCACGCTTCCTAGAGTTTGAAGCCCTAGGTTTCTTGAATGAAGACCACTGGTTCTCCAGAGAAAATTCACTGAGTGGAGTGGAAGGAGAAGGACTGCACAAGCTGGGATACATCCTAAGAGACATAAACAAGAAAGAAGGAGGAGCAATGTATGCTGATGACACAGCTGGTTGGGACACAAGAATCACAGAAGATGACCTGCAGAATGAAGAAATGATCACAGACCAAATGGACCCTGAACATAAGAAATTGGCCAAGGCCATTTTCAAACTAACATACCAAAACAAGGTGGTGAGAGTCCAAAGACCAACACCAAAAGGCACGGTGATGGATATCATATCCAGGAGAGACCAAAGAGGCAGTGGACAGGTTGGAACTTATGGTCTGAACACTTTCACCAACATGGAAGCACAACTAATCAGACAAATGGAAGGTGAAGGAGTCTTTACACCAAAAGAATTCTAGACCCCACATCTGCTAGAAAAAAGAGTTCAAGAATGGTTGGAAAAACATGGAGTCGAAAGGTTAAAAAGAATGGCAATCAGTGGAGATGATTGTGTGGTGAAACCACTAGATGACAGGTTCGCCACAGCCCTAACAGCTTTGAATGACATGGGAAAGGTTAGGAAAGACATACCGCAATGGGAACCTTCAAAAGGATGGAATGATTGGCAACAAGTGCCTTTCTGCTCCCACCACTTTCATGAGCTGATCATGAAAGATGGTCGCACGCTAGTAGTTCCATGCAGAAACCAAGATGAACTGATAGGTAGAGCCAGAATCTCTCAGGGAGCTGGATGGAGCTTGAGAGAAACAGCCTGTCTGGGGAAGTCTTACGCCCAAATGTGGAGGCTGATGTACTTCCACAGAAGAGATCTGAGGCTGGCGGCCAATGCCATATGTTCAGCAGTTCCAGTACATTGGGTTCCAACAAGCAGAACAACCTGGTCGATCCATGCCCACCATCAATGGATGACAACAGAAGACATGTTTACAGTCTGGAACAGGGTGTGGATAGAGGACAACCCATGGATGGAAGACAAAACTCCAGTCTAATCTTGGGAAGACATACCATACCTAGGGAAAAGAGAAGACCAGTGGTGTGGATCCCTGATTGGCCTAACATCCAGGGCCACCTGGGCAAAGAACATTCAAACAGCCATAAACCAAGTCAGAAGCCTCATTGGGAATGAGGAGTATATAGATTACATGCCATCAATGAAAAGATTCAGGAATGAGGAGGAGAACGAAGGAGTTCTGTGGTAAACAGCAAAAGAAGTCAGGCATAAAAGAAGTCAGGCCACTAATGCCACGGCTTGAGCAAACCGTGCTGCCTGTAGCTCCACCAAAAAGGGGAGGTGTAAAAAAACCCCGGGAGGCCACAAACCATGGAAGCTGTACGCATGGTGTAGTGGACTAGCGGTTAGAGGAGACCCCTCCCATGACAGACAAAACGCAGCACAAGGGGGCCCAAAGCCAGGAGGAAGCTGTACTCTCGTTGGAAGGACTAGAGGTTAGAGGAGACCCCCCCAAAACAAAAACAGCATATTGACGCTGGGAAAGACCAGAGATCCTGCTGTCTCCTCAGCATCATTCCAGGCACAGAACGCCAGAAAATGGAATGGTGCTGTTGAATCAACAGGTTCT