Rocafort-Gootenberg_Figure2ANCOM
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## vennlibrary("ggplotify")#Load original phyloseq object output from DADA2 pipeline and pull in new metadata
ps_gg_fp_f_prevalence_filter_2019_05_26<-readRDS("ps_gg_fp_f_prevalence_filter_2019_05_26")
readr::read_csv(
"Metadata_formatted_nat_comm_add_2021_10_24.csv",
col_names = TRUE,
col_types = NULL,
col_select = NULL,
id = NULL,
locale = default_locale(),
na = c("", "NA", "empty", "EMPTY"),
quote = "\"",
comment = "",
trim_ws = TRUE,
skip = 0,
name_repair = "unique",
num_threads = readr_threads(),
progress = show_progress(),
show_col_types = should_show_types(),
skip_empty_rows = TRUE,
lazy = TRUE
) -> new_metadata## Rows: 597 Columns: 88
## ── Column specification ────────────────────────────────────────────────────────
## Delimiter: ","
## chr (26): X, SampleID, subject_id, Race, Ethnicity, unique_id, sequencing_da...
## dbl (62): primer_used, read_count, age, height_cm, height_in, weight_kg, wei...
##
## ℹ Use `spec()` to retrieve the full column specification for this data.
## ℹ Specify the column types or set `show_col_types = FALSE` to quiet this message.### add {SampleID} as rownames
new_metadata_as_sample_data <- phyloseq::sample_data(new_metadata)
phyloseq::sample_names(new_metadata_as_sample_data) <- dplyr::pull(new_metadata, 1)
phyloseq::sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26) <- new_metadata_as_sample_data## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
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## Also defined by 'tidytree'#Fix randomness
set.seed(1)#Figure 2A
#FIGURE 2A
#--------------------------------------------------------------------------------------------------------------
#COMPARISON 1: NEG vs ART
#Transform count data in the phyloseq object
ps_gg_fp_f_prevalence_filter_2019_05_26_proportion<-phyloseq::transform_sample_counts(ps_gg_fp_f_prevalence_filter_2019_05_26, function(x)(x/sum(x)))## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'#Select samples of interest and update phyloseq object
metadata<-phyloseq::sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26)
metadata<-metadata[metadata$hiv_phenotype %in% c("1_hiv_negative","2_suppressed"),,drop=F]
metadata<-as.data.frame(as.matrix(metadata[metadata$sexual_orientation != "MSM",,drop=FALSE])) ### swap to dropping "MSM"
phyloseq::sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)<-metadata## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'#Run PCoA on the phyloseq object
ordination<-phyloseq::ordinate(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion, "PCoA", "unifrac")## Warning in matrix(tree$edge[order(tree$edge[, 1]), ][, 2], byrow = TRUE, : data
## length [8987] is not a sub-multiple or multiple of the number of rows [4494]ordination$values[1:2,]## Eigenvalues Relative_eig Rel_corr_eig Broken_stick Cum_corr_eig
## 1 8.666120 0.05178965 0.05085708 0.01560113 0.05085708
## 2 8.650009 0.05169337 0.05076261 0.01324819 0.10161970
## Cumul_br_stick
## 1 0.01560113
## 2 0.02884932metadata_ordered<-metadata[row.names(ordination$vectors),,drop=FALSE]
all.equal(row.names(metadata_ordered), row.names(ordination$vectors))## [1] TRUEmetadata_ordered$Unifrac1 <- ordination$vectors[,1]
metadata_ordered$Unifrac2 <- ordination$vectors[,2]
comparison1 <- metadata_ordered
comparison1$Grouping <- as.factor(paste(comparison1$hiv_phenotype, comparison1$sample_cohort, sep="_"))
#Let's plot the data
us<-comparison1[comparison1$sample_cohort == "boston",,drop=F]
botswana<-comparison1[comparison1$sample_cohort == "botswana",,drop=F]
uganda<-comparison1[comparison1$sample_cohort == "uganda_2",,drop=F]
plot_us_comparison<-ggplot2::ggplot(data=us, aes(x=Unifrac1, y=Unifrac2))+geom_point(color="royalblue4", aes(alpha=hiv_phenotype), size=2, shape=16)+
theme_bw()+stat_ellipse(color="royalblue4", aes(alpha=hiv_phenotype), size=1)+scale_alpha_manual(values=c(1,0.3))+ylim(c(-0.4, 0.4))+xlim(c(-0.4,0.5))+
ggtitle("neg-art boston")+geom_point(data=us %>% group_by(hiv_phenotype) %>% summarise_at(vars(matches("UniFrac")), mean),size=5, color="royalblue4", aes(alpha=hiv_phenotype))## Warning: Using `size` aesthetic for lines was deprecated in ggplot2 3.4.0.
## ℹ Please use `linewidth` instead.plot_botswana_comparison<-ggplot(data=botswana, aes(x=Unifrac1, y=Unifrac2))+geom_point(color="darkorange", aes(alpha=hiv_phenotype), size=2, shape=16)+
theme_bw()+stat_ellipse(color="darkorange", aes(alpha=hiv_phenotype), size=1)+scale_alpha_manual(values=c(1,0.3))+ylim(c(-0.4, 0.4))+xlim(c(-0.4,0.5))+
ggtitle("neg-art botswana")+geom_point(data=botswana %>% group_by(hiv_phenotype) %>% summarise_at(vars(matches("UniFrac")), mean),size=5, color="darkorange", aes(alpha=hiv_phenotype))
plot_uganda_comparison<-ggplot(data=uganda, aes(x=Unifrac1, y=Unifrac2))+geom_point(color="forestgreen", aes(alpha=hiv_phenotype), size=2, shape=16)+
theme_bw()+stat_ellipse(color="forestgreen", aes(alpha=hiv_phenotype), size=1)+scale_alpha_manual(values=c(1,0.3))+ylim(c(-0.4, 0.4))+xlim(c(-0.4,0.5))+
ggtitle("neg-art uganda")+geom_point(data=uganda %>% group_by(hiv_phenotype) %>% summarise_at(vars(matches("UniFrac")), mean),size=5, color="forestgreen", aes(alpha=hiv_phenotype))
ggsave("Figure2A_1v3.pdf", grid.arrange(plot_us_comparison,plot_botswana_comparison,plot_uganda_comparison, nrow=3, ncol=3), width=15, height=10)
#Adonis (n = 427)
ASV_table <- as.data.frame(phyloseq::otu_table(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion))
all.equal(row.names(ASV_table), row.names(metadata_ordered))## [1] TRUEunifrac.distance <- unname(phyloseq::UniFrac(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion, weighted = FALSE)) ### unname fixes error introduced by Desctools see https://github.com/joey711/phyloseq/issues/1457## Warning in matrix(tree$edge[order(tree$edge[, 1]), ][, 2], byrow = TRUE, : data
## length [8987] is not a sub-multiple or multiple of the number of rows [4494]attributes(unifrac.distance)$Labels <- phyloseq::sample_names(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)
print(vegan::adonis2(unifrac.distance ~ metadata_ordered$hiv_phenotype, data = ASV_table, permutations = 1000)) -> adon_hiv_phenotype## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = unifrac.distance ~ metadata_ordered$hiv_phenotype, data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$hiv_phenotype 1 1.148 0.00686 2.9357 0.000999 ***
## Residual 425 166.185 0.99314
## Total 426 167.333 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** hiv_phenotype r2 = 0.00686 p = 0.000999
print(vegan::adonis2(unifrac.distance ~ metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)) -> adon_cohort## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = unifrac.distance ~ metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$sample_cohort 2 8.094 0.04837 10.776 0.000999 ***
## Residual 424 159.239 0.95163
## Total 426 167.333 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.04837 p = 0.000999
print(vegan::adonis2(unifrac.distance ~ metadata_ordered$hiv_phenotype + metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)) -> adon_cohort_phenotype## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = unifrac.distance ~ metadata_ordered$hiv_phenotype + metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$hiv_phenotype 1 1.148 0.00686 3.0702 0.000999 ***
## metadata_ordered$sample_cohort 2 8.026 0.04797 10.7332 0.000999 ***
## Residual 423 158.159 0.94517
## Total 426 167.333 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.04797 p = 0.000999, hiv_phenotype r2 = 0.00686 p = 0.000999
#--------------------------------------------------------------------------------------------------------------
### COMPARISON 1: Adonis: controlling for additional metadata
#--------------------------------------------------------------------------------------------------------------
### Extra metadata that have full n: Race, Ethnicity, age, sex, current_art_class_consolid2, tmp_smx_active
### Extra metadata that have <n: BMI, comorbidities (hld_hx, htn_hx, cvd_hx, ckd_hx, cvd_dx [missing boston], dm2hx_dx, hldhx_dx, htnhx_dx, cvdhx_dx, ever_smoke, current_smoke, smoke_cat), school_level [uganda2 only], monthly_income[uganda2 only], current_art_class_consolid2, tmp_smx_active, days_on_art, sexual_orientation
### additional: smoking_years, fram_10yr_risk_lab, fram_10yr_risk_nonlab, mean_imt, total_plaques, any_plaques
metadata_ordered$age <- as.numeric(metadata_ordered$age)
metadata_ordered$BMI <- as.numeric(metadata_ordered$BMI)
metadata_ordered$days_on_art <- as.numeric(metadata_ordered$days_on_art)
### control for metadata with full n
covars_full_n <- c("Ethnicity", "age", "sex", "Race", "hiv_phenotype", "tmp_smx_active", "current_art_class_consolid2", "sample_cohort")
print(vegan::adonis2(as.formula(paste("unifrac.distance~metadata_ordered$", paste(covars_full_n, collapse = "+metadata_ordered$"), sep = "")), data=ASV_table, permutations=1000)) -> adon_full_n## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = as.formula(paste("unifrac.distance~metadata_ordered$", paste(covars_full_n, collapse = "+metadata_ordered$"), sep = "")), data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F
## metadata_ordered$Ethnicity 1 0.489 0.00292 1.3177
## metadata_ordered$age 1 1.448 0.00865 3.9011
## metadata_ordered$sex 1 0.682 0.00408 1.8372
## metadata_ordered$Race 5 5.128 0.03064 2.7626
## metadata_ordered$hiv_phenotype 1 1.064 0.00636 2.8659
## metadata_ordered$tmp_smx_active 1 1.568 0.00937 4.2230
## metadata_ordered$current_art_class_consolid2 3 1.486 0.00888 1.3346
## metadata_ordered$sample_cohort 2 2.898 0.01732 3.9032
## Residual 411 152.570 0.91178
## Total 426 167.333 1.00000
## Pr(>F)
## metadata_ordered$Ethnicity 0.026973 *
## metadata_ordered$age 0.000999 ***
## metadata_ordered$sex 0.000999 ***
## metadata_ordered$Race 0.000999 ***
## metadata_ordered$hiv_phenotype 0.000999 ***
## metadata_ordered$tmp_smx_active 0.000999 ***
## metadata_ordered$current_art_class_consolid2 0.001998 **
## metadata_ordered$sample_cohort 0.000999 ***
## Residual
## Total
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.01732 p = 0.000999, hiv_phenotype r2 = 0.00636 p = 0.000999
# Df SumOfSqs R2 F Pr(>F)
# metadata_ordered$Ethnicity 1 0.489 0.00292 1.3177 0.023976 *
# metadata_ordered$age 1 1.448 0.00865 3.9011 0.000999 ***
# metadata_ordered$sex 1 0.682 0.00408 1.8372 0.000999 ***
# metadata_ordered$Race 5 5.128 0.03064 2.7626 0.000999 ***
# metadata_ordered$hiv_phenotype 1 1.064 0.00636 2.8659 0.000999 ***
# metadata_ordered$tmp_smx_active 1 1.568 0.00937 4.2230 0.000999 ***
# metadata_ordered$current_art_class_consolid2 3 1.486 0.00888 1.3346 0.000999 ***
# metadata_ordered$sample_cohort 2 2.898 0.01732 3.9032 0.000999 ***
# Residual 411 152.570 0.91178
# Total 426 167.333 1.00000
#--------------------------------------------------------------------------------------------------------------
### COMPARISON 1: Adonis: controlling for additional metadata - removing samples w/o values - BMI/comorbidities/days_on_art
#--------------------------------------------------------------------------------------------------------------
### remove samples with BMI/comorbidities/days_on_art is NA (n = 427 -> n = 405)
ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bcd <- ps_gg_fp_f_prevalence_filter_2019_05_26_proportion
metadata_ordered_bcd <- metadata_ordered[!is.na(metadata_ordered$hld_hx) & !is.na(metadata_ordered$days_on_art) & !is.na(metadata_ordered$BMI), , drop = FALSE]
sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bcd) <- metadata_ordered_bcd## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'### subset ASV table and distance matrix
ASV_table_bcd <- as.data.frame(otu_table(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bcd))
all.equal(row.names(ASV_table_bcd), row.names(metadata_ordered_bcd))## [1] TRUEunifrac.distance_bcd <- unname(usedist::dist_subset(unifrac.distance, sample_names(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bcd))) ### unname fixes error introduced by Desctools see https://github.com/joey711/phyloseq/issues/1457
attributes(unifrac.distance_bcd)$Labels <- phyloseq::sample_names(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bcd)
### run adonis
covars_bcd <- c("BMI", "hld_hx", "htn_hx", "cvd_hx", "dm2hx_dx", "hldhx_dx", "cvdhx_dx", "days_on_art", covars_full_n)
print(vegan::adonis2(as.formula(paste("unifrac.distance_bcd~metadata_ordered_bcd$", paste(covars_bcd, collapse = "+metadata_ordered_bcd$"), sep = "")), data = ASV_table_bcd, permutations=1000)) -> adon_full_n_bcd## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = as.formula(paste("unifrac.distance_bcd~metadata_ordered_bcd$", paste(covars_bcd, collapse = "+metadata_ordered_bcd$"), sep = "")), data = ASV_table_bcd, permutations = 1000)
## Df SumOfSqs R2 F
## metadata_ordered_bcd$BMI 1 0.983 0.00621 2.6552
## metadata_ordered_bcd$hld_hx 1 0.457 0.00289 1.2361
## metadata_ordered_bcd$htn_hx 1 0.515 0.00326 1.3917
## metadata_ordered_bcd$cvd_hx 1 0.622 0.00393 1.6796
## metadata_ordered_bcd$dm2hx_dx 1 0.443 0.00280 1.1978
## metadata_ordered_bcd$hldhx_dx 1 0.534 0.00338 1.4439
## metadata_ordered_bcd$cvdhx_dx 1 0.684 0.00433 1.8494
## metadata_ordered_bcd$days_on_art 1 0.928 0.00587 2.5080
## metadata_ordered_bcd$Ethnicity 1 0.487 0.00308 1.3146
## metadata_ordered_bcd$age 1 1.184 0.00749 3.1999
## metadata_ordered_bcd$sex 1 0.798 0.00504 2.1563
## metadata_ordered_bcd$Race 4 3.610 0.02282 2.4388
## metadata_ordered_bcd$hiv_phenotype 1 0.731 0.00462 1.9738
## metadata_ordered_bcd$tmp_smx_active 1 1.008 0.00637 2.7248
## metadata_ordered_bcd$current_art_class_consolid2 3 1.554 0.00982 1.3996
## metadata_ordered_bcd$sample_cohort 2 2.292 0.01449 3.0969
## Residual 382 141.381 0.89361
## Total 404 158.214 1.00000
## Pr(>F)
## metadata_ordered_bcd$BMI 0.000999 ***
## metadata_ordered_bcd$hld_hx 0.068931 .
## metadata_ordered_bcd$htn_hx 0.013986 *
## metadata_ordered_bcd$cvd_hx 0.000999 ***
## metadata_ordered_bcd$dm2hx_dx 0.058941 .
## metadata_ordered_bcd$hldhx_dx 0.005994 **
## metadata_ordered_bcd$cvdhx_dx 0.000999 ***
## metadata_ordered_bcd$days_on_art 0.000999 ***
## metadata_ordered_bcd$Ethnicity 0.030969 *
## metadata_ordered_bcd$age 0.000999 ***
## metadata_ordered_bcd$sex 0.000999 ***
## metadata_ordered_bcd$Race 0.000999 ***
## metadata_ordered_bcd$hiv_phenotype 0.000999 ***
## metadata_ordered_bcd$tmp_smx_active 0.000999 ***
## metadata_ordered_bcd$current_art_class_consolid2 0.000999 ***
## metadata_ordered_bcd$sample_cohort 0.000999 ***
## Residual
## Total
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.01449 p = 0.000999, hiv_phenotype r2 = 0.00462 p = 0.000999
# Df SumOfSqs R2 F Pr(>F)
# metadata_ordered_bcd$BMI 1 0.983 0.00621 2.6552 0.000999 ***
# metadata_ordered_bcd$hld_hx 1 0.457 0.00289 1.2361 0.057942 .
# metadata_ordered_bcd$htn_hx 1 0.515 0.00326 1.3917 0.008991 **
# metadata_ordered_bcd$cvd_hx 1 0.622 0.00393 1.6796 0.000999 ***
# metadata_ordered_bcd$dm2hx_dx 1 0.443 0.00280 1.1978 0.089910 .
# metadata_ordered_bcd$hldhx_dx 1 0.534 0.00338 1.4439 0.007992 **
# metadata_ordered_bcd$cvdhx_dx 1 0.684 0.00433 1.8494 0.000999 ***
# metadata_ordered_bcd$days_on_art 1 0.928 0.00587 2.5080 0.000999 ***
# metadata_ordered_bcd$Ethnicity 1 0.487 0.00308 1.3146 0.024975 *
# metadata_ordered_bcd$age 1 1.184 0.00749 3.1999 0.000999 ***
# metadata_ordered_bcd$sex 1 0.798 0.00504 2.1563 0.000999 ***
# metadata_ordered_bcd$Race 4 3.610 0.02282 2.4388 0.000999 ***
# metadata_ordered_bcd$hiv_phenotype 1 0.731 0.00462 1.9738 0.000999 ***
# metadata_ordered_bcd$tmp_smx_active 1 1.008 0.00637 2.7248 0.000999 ***
# metadata_ordered_bcd$current_art_class_consolid2 3 1.554 0.00982 1.3996 0.000999 ***
# metadata_ordered_bcd$sample_cohort 2 2.292 0.01449 3.0969 0.000999 ***
# Residual 382 141.381 0.89361
# Total 404 158.214 1.00000
#--------------------------------------------------------------------------------------------------------------
#--------------------------------------------------------------------------------------------------------------
#COMPARISON 2: NEG vs UNTREATED
#Transform count data in the phyloseq object
ps_gg_fp_f_prevalence_filter_2019_05_26_proportion <- transform_sample_counts(ps_gg_fp_f_prevalence_filter_2019_05_26, function(x)(x/sum(x)))## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'#Select samples of interest and update phyloseq object
metadata <- phyloseq::sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26)
metadata <- metadata[metadata$hiv_phenotype%in%c("1_hiv_negative","4_unsuppressed"),,drop=F]
metadata <- as.data.frame(as.matrix(metadata[metadata$sexual_orientation!="MSM",,drop=F]))
metadata <- metadata[metadata$sample_cohort%in%c("botswana", "boston"),,drop=F]
phyloseq::sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)<-metadata## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'#Run PCoA on the phyloseq object
ordination <- phyloseq::ordinate(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion, "PCoA", "unifrac")## Warning in matrix(tree$edge[order(tree$edge[, 1]), ][, 2], byrow = TRUE, : data
## length [8987] is not a sub-multiple or multiple of the number of rows [4494]ordination$values[1:2,]## Eigenvalues Relative_eig Broken_stick Cumul_eig Cumul_br_stick
## 1 5.257069 0.06368984 0.02756855 0.06368984 0.02756855
## 2 4.098253 0.04965069 0.02293892 0.11334053 0.05050747metadata_ordered<-metadata[row.names(ordination$vectors),,drop=FALSE]
all.equal(row.names(metadata_ordered), row.names(ordination$vectors))## [1] TRUEmetadata_ordered$Unifrac1 <- ordination$vectors[,1]
metadata_ordered$Unifrac2 <- ordination$vectors[,2]
comparison2 <- metadata_ordered
comparison2$Grouping <- as.factor(paste(comparison2$hiv_phenotype, comparison2$sample_cohort, sep="_"))
#Let's plot the data
us<-comparison2[comparison2$sample_cohort=="boston",,drop=F]
botswana<-comparison2[comparison2$sample_cohort=="botswana",,drop=F]
plot_us_comparison<-ggplot2::ggplot(data=us, aes(x=Unifrac1, y=Unifrac2))+geom_point(color="royalblue4", aes(alpha=hiv_phenotype), size=2, shape=16)+
theme_bw()+stat_ellipse(color="royalblue4", aes(alpha=hiv_phenotype), size=1)+scale_alpha_manual(values=c(1,0.3))+ylim(c(-0.5, 0.5))+xlim(c(-0.5,0.5))+
ggtitle("neg-naive us")+geom_point(data=us %>% group_by(hiv_phenotype) %>% summarise_at(vars(matches("UniFrac")), mean),size=5, color="royalblue4", aes(alpha=hiv_phenotype))
plot_botswana_comparison<-ggplot(data=botswana, aes(x=Unifrac1, y=Unifrac2))+geom_point(color="darkorange", aes(alpha=hiv_phenotype), size=2, shape=16)+
theme_bw()+stat_ellipse(color="darkorange", aes(alpha=hiv_phenotype), size=1)+scale_alpha_manual(values=c(1,0.3))+ylim(c(-0.5, 0.5))+xlim(c(-0.5,0.5))+
ggtitle("neg-naive botswana")+geom_point(data=botswana %>% group_by(hiv_phenotype) %>% summarise_at(vars(matches("UniFrac")), mean),size=5, color="darkorange", aes(alpha=hiv_phenotype))
ggsave("Figure2A_2v3.pdf", grid.arrange(plot_us_comparison,plot_botswana_comparison, nrow=3, ncol=3), width=15, height=10)
#Adonis (n = 217)
ASV_table <- as.data.frame(phyloseq::otu_table(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion))
all.equal(row.names(ASV_table), row.names(metadata_ordered))## [1] TRUEunifrac.distance <- unname(phyloseq::UniFrac(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion, weighted = FALSE)) ### unname fixes error introduced by Desctools see https://github.com/joey711/phyloseq/issues/1457## Warning in matrix(tree$edge[order(tree$edge[, 1]), ][, 2], byrow = TRUE, : data
## length [8987] is not a sub-multiple or multiple of the number of rows [4494]attributes(unifrac.distance)$Labels <- phyloseq::sample_names(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)
print(vegan::adonis2(unifrac.distance ~ metadata_ordered$hiv_phenotype, data = ASV_table, permutations = 1000)) -> adon_hiv_phenotype## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = unifrac.distance ~ metadata_ordered$hiv_phenotype, data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$hiv_phenotype 1 0.550 0.00666 1.4412 0.008991 **
## Residual 215 81.992 0.99334
## Total 216 82.542 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** r2 = 0.00666 p = 0.005994
print(vegan::adonis2(unifrac.distance ~ metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)) -> adon_cohort## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = unifrac.distance ~ metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$sample_cohort 1 2.805 0.03398 7.563 0.000999 ***
## Residual 215 79.737 0.96602
## Total 216 82.542 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** r2 = 0.03398 p = 0.000999
print(vegan::adonis2(unifrac.distance ~ metadata_ordered$hiv_phenotype + metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)) -> adon_cohort_phenotype## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = unifrac.distance ~ metadata_ordered$hiv_phenotype + metadata_ordered$sample_cohort, data = ASV_table, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$hiv_phenotype 1 0.550 0.00666 1.4840 0.007992 **
## metadata_ordered$sample_cohort 1 2.736 0.03315 7.3888 0.000999 ***
## Residual 214 79.256 0.96019
## Total 216 82.542 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.03315 p = 0.000999, hiv_phenotype r2 = 0.00666 p = 0.008991
#--------------------------------------------------------------------------------------------------------------
### COMPARISON 2: Adonis: controlling for additional metadata
#--------------------------------------------------------------------------------------------------------------
### Extra metadata to add: (in addition to sample_cohort, hiv_phenotype)
### that have full n: Race, Ethnicity, age, sex
metadata_ordered$age <- as.numeric(metadata_ordered$age)
metadata_ordered$BMI <- as.numeric(metadata_ordered$BMI)
### metadata not collected in Boston:
metadata_ordered$monthly_income <- as.numeric(metadata_ordered$monthly_income)
metadata_ordered$smoking_years <- as.numeric(metadata_ordered$smoking_years)
metadata_ordered$fram_10yr_risk_lab <- as.numeric(metadata_ordered$fram_10yr_risk_lab)
metadata_ordered$fram_10yr_risk_nonlab <- as.numeric(metadata_ordered$fram_10yr_risk_nonlab)
metadata_ordered$mean_imt <- as.numeric(metadata_ordered$mean_imt)
metadata_ordered$total_plaques <- as.numeric(metadata_ordered$total_plaques)
metadata_ordered$any_plaques <- as.numeric(metadata_ordered$any_plaques)
### control for metadata with full n
covars_full_n_untreat <- covars_full_n[!covars_full_n %in% c("tmp_smx_active", "current_art_class_consolid2")]
print(vegan::adonis2(as.formula(paste("unifrac.distance~metadata_ordered$", paste(covars_full_n_untreat, collapse = "+metadata_ordered$"), sep = "")), data=ASV_table, permutations=1000, by = "margin")) -> adon_full_n_marg## Permutation test for adonis under reduced model
## Marginal effects of terms
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = as.formula(paste("unifrac.distance~metadata_ordered$", paste(covars_full_n_untreat, collapse = "+metadata_ordered$"), sep = "")), data = ASV_table, permutations = 1000, by = "margin")
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered$Ethnicity 1 0.342 0.00414 0.9289 0.668332
## metadata_ordered$age 1 0.604 0.00732 1.6423 0.003996 **
## metadata_ordered$sex 1 0.442 0.00536 1.2025 0.073926 .
## metadata_ordered$Race 4 1.708 0.02069 1.1615 0.018981 *
## metadata_ordered$hiv_phenotype 1 0.471 0.00571 1.2816 0.041958 *
## metadata_ordered$sample_cohort 1 0.848 0.01027 2.3059 0.000999 ***
## Residual 207 76.105 0.92202
## Total 216 82.542 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.01027 p = 0.000999, hiv_phenotype r2 = 0.00571 p = 0.040959
# Df SumOfSqs R2 F Pr(>F)
# metadata_ordered$Ethnicity 1 0.342 0.00414 0.9289 0.683317
# metadata_ordered$age 1 0.604 0.00732 1.6423 0.002997 **
# metadata_ordered$sex 1 0.442 0.00536 1.2025 0.095904 .
# metadata_ordered$Race 4 1.708 0.02069 1.1615 0.021978 *
# metadata_ordered$hiv_phenotype 1 0.471 0.00571 1.2816 0.040959 *
# metadata_ordered$sample_cohort 1 0.848 0.01027 2.3059 0.000999 ***
# Residual 207 76.105 0.92202
# Total 216 82.542 1.00000
#--------------------------------------------------------------------------------------------------------------
### COMPARISON 2: Adonis: controlling for additional metadata - removing samples w/o values - BMI/comorbidities
#--------------------------------------------------------------------------------------------------------------
### remove samples with BMI/comorbidities is NA (n=217 -> n=201)
ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bc <- ps_gg_fp_f_prevalence_filter_2019_05_26_proportion
metadata_ordered_bc <- metadata_ordered[!is.na(metadata_ordered$hld_hx) & !is.na(metadata_ordered$BMI), , drop = FALSE]
sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bc) <- metadata_ordered_bc## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'### subset ASV table and distance matrix
ASV_table_bc<-as.data.frame(otu_table(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bc))
all.equal(row.names(ASV_table_bc), row.names(metadata_ordered_bc))## [1] TRUEunifrac.distance_bc <- unname(usedist::dist_subset(unifrac.distance, sample_names(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bc))) ### unname fixes error introduced by Desctools see https://github.com/joey711/phyloseq/issues/1457
attributes(unifrac.distance_bc)$Labels <- phyloseq::sample_names(ps_gg_fp_f_prevalence_filter_2019_05_26_proportion_bc)
### run adonis
covars_bc <- c("BMI", "hld_hx", "htn_hx", "cvd_hx", "dm2hx_dx", "hldhx_dx", "cvdhx_dx", covars_full_n_untreat)
print(vegan::adonis2(as.formula(paste("unifrac.distance_bc~metadata_ordered_bc$", paste(covars_bc, collapse = "+metadata_ordered_bc$"), sep = "")), data=ASV_table_bc, permutations=1000)) -> adon_full_n_bc## Permutation test for adonis under reduced model
## Terms added sequentially (first to last)
## Permutation: free
## Number of permutations: 1000
##
## vegan::adonis2(formula = as.formula(paste("unifrac.distance_bc~metadata_ordered_bc$", paste(covars_bc, collapse = "+metadata_ordered_bc$"), sep = "")), data = ASV_table_bc, permutations = 1000)
## Df SumOfSqs R2 F Pr(>F)
## metadata_ordered_bc$BMI 1 0.542 0.00713 1.4843 0.010989 *
## metadata_ordered_bc$hld_hx 1 0.432 0.00568 1.1830 0.109890
## metadata_ordered_bc$htn_hx 1 0.360 0.00474 0.9874 0.456543
## metadata_ordered_bc$cvd_hx 1 0.502 0.00660 1.3755 0.010989 *
## metadata_ordered_bc$dm2hx_dx 1 0.397 0.00522 1.0880 0.204795
## metadata_ordered_bc$hldhx_dx 1 0.597 0.00785 1.6350 0.001998 **
## metadata_ordered_bc$cvdhx_dx 1 0.511 0.00672 1.4002 0.010989 *
## metadata_ordered_bc$Ethnicity 1 0.413 0.00543 1.1309 0.164835
## metadata_ordered_bc$age 1 0.879 0.01156 2.4075 0.000999 ***
## metadata_ordered_bc$sex 1 0.451 0.00593 1.2354 0.066933 .
## metadata_ordered_bc$Race 4 2.699 0.03550 1.8488 0.000999 ***
## metadata_ordered_bc$hiv_phenotype 1 0.423 0.00557 1.1599 0.104895
## metadata_ordered_bc$sample_cohort 1 0.660 0.00869 1.8091 0.000999 ***
## Residual 184 67.155 0.88338
## Total 200 76.020 1.00000
## ---
## Signif. codes: 0 '***' 0.001 '**' 0.01 '*' 0.05 '.' 0.1 ' ' 1###*** sample_cohort r2 = 0.00854 p = 0.000999, hiv_phenotype r2 = 0.00557 p = 0.135864
# Df SumOfSqs R2 F Pr(>F)
# metadata_ordered_bc$BMI 1 0.542 0.00713 1.4843 0.006993 **
# metadata_ordered_bc$hld_hx 1 0.432 0.00568 1.1830 0.118881
# metadata_ordered_bc$htn_hx 1 0.360 0.00474 0.9874 0.481518
# metadata_ordered_bc$cvd_hx 1 0.502 0.00660 1.3755 0.015984 *
# metadata_ordered_bc$dm2hx_dx 1 0.397 0.00522 1.0880 0.210789
# metadata_ordered_bc$hldhx_dx 1 0.597 0.00785 1.6350 0.000999 ***
# metadata_ordered_bc$cvdhx_dx 1 0.511 0.00672 1.4002 0.014985 *
# metadata_ordered_bc$Ethnicity 1 0.413 0.00543 1.1309 0.135864
# metadata_ordered_bc$age 1 0.879 0.01156 2.4075 0.000999 ***
# metadata_ordered_bc$sex 1 0.451 0.00593 1.2354 0.061938 .
# metadata_ordered_bc$Race 4 2.699 0.03550 1.8488 0.000999 ***
# metadata_ordered_bc$hiv_phenotype 1 0.423 0.00557 1.1599 0.119880
# metadata_ordered_bc$sample_cohort 1 0.660 0.00869 1.8091 0.001998 **
# Residual 184 67.155 0.88338
# Total 200 76.020 1.00000
#--------------------------------------------------------------------------------------------------------------#Figure 2BCDE
#FIGURE 2B, C, D: ANCOM NEG-ART & FIGURE 2E
#--------------------------------------------------------------------------------------------------------------
#Transform count data in the phyloseq object
ps_gg_fp_f_prevalence_filter_2019_05_26_proportion<-transform_sample_counts(ps_gg_fp_f_prevalence_filter_2019_05_26, function(x)(x/sum(x)))## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'###RUN ANCOM###
#US
dataset<-ps_gg_fp_f_prevalence_filter_2019_05_26
metadata<-as.data.frame(sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26))
metadata<-metadata[metadata$hiv_phenotype %in% c("1_hiv_negative", "2_suppressed"), , drop=F]
metadata<-as.data.frame(as.matrix(metadata[metadata$sexual_orientation != "MSM" | is.na(metadata$sexual_orientation), , drop=F]))
metadata_boston<-metadata[metadata$sample_cohort == "boston", , drop=F]
sample_data(dataset)<-metadata_boston## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'ps.taxa.sub <- phyloseq::prune_taxa(taxa_sums(dataset) > 0, dataset)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'out <- ANCOMBC::ancombc(data = ps.taxa.sub, formula = "hiv_phenotype", tax_level = NULL,
p_adj_method = "BH", prv_cut = 0.05, lib_cut = 1000,
group = "hiv_phenotype", struc_zero = TRUE, neg_lb = FALSE, tol = 1e-5,
max_iter = 100, conserve = FALSE, alpha = 0.05, global = FALSE, n_cl = 6)## 'ancombc' is deprecated
## Use 'ancombc2' instead## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## `tax_level` is not speficified
## No agglomeration will be performed
## Otherwise, please speficy `tax_level` by one of the following:
## Kingdom, Phylum, Class, Order, Family, Genus, Species## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Warning: The group variable has < 3 categories
## The multi-group comparisons (global/pairwise/dunnet/trend) will be deactivated## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'res <- out$res
res_rn <- purrr::imap(res, function(x, y) dplyr::rename(x, !!y := hiv_phenotype2_suppressed))
res_df <- purrr::reduce(res_rn, dplyr::left_join, by = "taxon")
res_df <- dplyr::select(res_df, !starts_with("(Int"))
res_df_taxa <- dplyr::left_join(res_df, tibble::rownames_to_column(as.data.frame(phyloseq::tax_table(ps.taxa.sub))), by = c("taxon" = "rowname"))
res_df_taxa[["index_num"]] <- 1:nrow(res_df_taxa)
res_df_taxa[["cohort"]] <- "boston"
res_df_taxa[["method"]] <- "ancom"
res_df_taxa <- tidyr::unite(res_df_taxa, col = "Genus_Species", Genus, Species, index_num, remove = FALSE)
alpha = 0.05
taxa_sig <- dplyr::filter(res_df_taxa, q_val < 0.05)## Warning: Using one column matrices in `filter()` was deprecated in dplyr 1.1.0.
## ℹ Please use one dimensional logical vectors instead.
## ℹ The deprecated feature was likely used in the dplyr package.
## Please report the issue at <]8;;https://github.com/tidyverse/dplyr/issueshttps://github.com/tidyverse/dplyr/issues]8;;>.taxa_sig$Genus_Species <- forcats::fct_reorder(taxa_sig$Genus_Species, taxa_sig$lfc, min)
taxa_sig$taxon_short <- stringr::str_sub(taxa_sig$taxon, 1, 4)
ps.taxa.rel.sig <- phyloseq::prune_taxa(taxa_sig[["taxon"]], ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'# Only keep filtered samples
ps.taxa.rel.sig <- phyloseq::prune_samples(rownames(phyloseq::otu_table(ps.taxa.sub)), ps.taxa.rel.sig)
sigtab_dataset_us <- taxa_sig
write.csv(sigtab_dataset_us, "ANCOM_US_NEG_ART_Filtered.csv")
#BOTSWANA
dataset<-ps_gg_fp_f_prevalence_filter_2019_05_26
metadata<-as.data.frame(sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26))
metadata<-metadata[metadata$hiv_phenotype %in% c("1_hiv_negative", "2_suppressed"), , drop=F]
metadata<-as.data.frame(as.matrix(metadata[metadata$sexual_orientation != "MSM" | is.na(metadata$sexual_orientation), , drop=F]))
metadata_botswana<-metadata[metadata$sample_cohort == "botswana", , drop=F]
sample_data(dataset)<-metadata_botswana## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'ps.taxa.sub <- phyloseq::prune_taxa(taxa_sums(dataset) > 0, dataset)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'out <- ANCOMBC::ancombc(data = ps.taxa.sub, formula = "hiv_phenotype", tax_level = NULL,
p_adj_method = "BH", prv_cut = 0.05, lib_cut = 1000,
group = "hiv_phenotype", struc_zero = TRUE, neg_lb = FALSE, tol = 1e-5,
max_iter = 100, conserve = FALSE, alpha = 0.05, global = FALSE, n_cl = 6)## 'ancombc' is deprecated
## Use 'ancombc2' instead## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## `tax_level` is not speficified
## No agglomeration will be performed
## Otherwise, please speficy `tax_level` by one of the following:
## Kingdom, Phylum, Class, Order, Family, Genus, Species## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Warning: The group variable has < 3 categories
## The multi-group comparisons (global/pairwise/dunnet/trend) will be deactivated## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'res <- out$res
res_rn <- purrr::imap(res, function(x, y) dplyr::rename(x, !!y := hiv_phenotype2_suppressed))
res_df <- purrr::reduce(res_rn, dplyr::left_join, by = "taxon")
res_df <- dplyr::select(res_df, !starts_with("(Int"))
res_df_taxa <- dplyr::left_join(res_df, tibble::rownames_to_column(as.data.frame(phyloseq::tax_table(ps.taxa.sub))), by = c("taxon" = "rowname"))
res_df_taxa[["index_num"]] <- 1:nrow(res_df_taxa)
res_df_taxa[["cohort"]] <- "botswana"
res_df_taxa[["method"]] <- "ancom"
res_df_taxa <- tidyr::unite(res_df_taxa, col = "Genus_Species", Genus, Species, index_num, remove = FALSE)
alpha = 0.05
taxa_sig <- dplyr::filter(res_df_taxa, q_val < 0.05)
taxa_sig$Genus_Species <- forcats::fct_reorder(taxa_sig$Genus_Species, taxa_sig$lfc, min)
taxa_sig$taxon_short <- stringr::str_sub(taxa_sig$taxon, 1, 4)
ps.taxa.rel.sig <- phyloseq::prune_taxa(taxa_sig[["taxon"]], ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'# Only keep filtered samples
ps.taxa.rel.sig <- phyloseq::prune_samples(rownames(phyloseq::otu_table(ps.taxa.sub)), ps.taxa.rel.sig)
sigtab_dataset_botswana <- taxa_sig
write.csv(sigtab_dataset_botswana, "ANCOM_BOTS_NEG_ART_Filtered.csv")
#UGANDA
dataset<-ps_gg_fp_f_prevalence_filter_2019_05_26
metadata<-as.data.frame(sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26))
metadata<-metadata[metadata$hiv_phenotype %in% c("1_hiv_negative", "2_suppressed"), , drop=F]
metadata<-as.data.frame(as.matrix(metadata[metadata$sexual_orientation != "MSM" | is.na(metadata$sexual_orientation), , drop=F]))
metadata_uganda<-metadata[metadata$sample_cohort == "uganda_2", , drop=F]
sample_data(dataset)<-metadata_uganda## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'ps.taxa.sub <- phyloseq::prune_taxa(taxa_sums(dataset) > 0, dataset)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'out <- ANCOMBC::ancombc(data = ps.taxa.sub, formula = "hiv_phenotype", tax_level = NULL,
p_adj_method = "BH", prv_cut = 0.05, lib_cut = 1000,
group = "hiv_phenotype", struc_zero = TRUE, neg_lb = FALSE, tol = 1e-5,
max_iter = 100, conserve = FALSE, alpha = 0.05, global = FALSE, n_cl = 6)## 'ancombc' is deprecated
## Use 'ancombc2' instead## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## `tax_level` is not speficified
## No agglomeration will be performed
## Otherwise, please speficy `tax_level` by one of the following:
## Kingdom, Phylum, Class, Order, Family, Genus, Species## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Warning: The group variable has < 3 categories
## The multi-group comparisons (global/pairwise/dunnet/trend) will be deactivated## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'res <- out$res
res_rn <- purrr::imap(res, function(x, y) dplyr::rename(x, !!y := hiv_phenotype2_suppressed))
res_df <- purrr::reduce(res_rn, dplyr::left_join, by = "taxon")
res_df <- dplyr::select(res_df, !starts_with("(Int"))
res_df_taxa <- dplyr::left_join(res_df, tibble::rownames_to_column(as.data.frame(phyloseq::tax_table(ps.taxa.sub))), by = c("taxon" = "rowname"))
res_df_taxa[["index_num"]] <- 1:nrow(res_df_taxa)
res_df_taxa[["cohort"]] <- "uganda_2"
res_df_taxa[["method"]] <- "ancom"
res_df_taxa <- tidyr::unite(res_df_taxa, col = "Genus_Species", Genus, Species, index_num, remove = FALSE)
alpha = 0.05
taxa_sig <- dplyr::filter(res_df_taxa, q_val < 0.05)
taxa_sig$Genus_Species <- forcats::fct_reorder(taxa_sig$Genus_Species, taxa_sig$lfc, min)
taxa_sig$taxon_short <- stringr::str_sub(taxa_sig$taxon, 1, 4)
ps.taxa.rel.sig <- phyloseq::prune_taxa(taxa_sig[["taxon"]], ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'# Only keep filtered samples
ps.taxa.rel.sig <- phyloseq::prune_samples(rownames(phyloseq::otu_table(ps.taxa.sub)), ps.taxa.rel.sig)
sigtab_dataset_uganda <- taxa_sig
write.csv(sigtab_dataset_uganda, "ANCOM_UGANDA_NEG_ART_Filtered.csv")
# Save plots of individual ANCOM fold-change
ggsave("Figure2BCD_NEG_ART_ANCOM_US_v7.pdf", ggplot(data = sigtab_dataset_us, aes(x = Genus_Species, y = lfc)) + theme_bw() + coord_flip() + geom_bar(stat="identity", aes(fill = Genus)) + ggtitle("NEG-ART US") + theme(legend.position = "bottom"), width = 10, height = 10, units = "in", dpi = 300)
ggsave("Figure2BCD_NEG_ART_ANCOM_BOTS_v7.pdf", ggplot(data = sigtab_dataset_botswana, aes(x = Genus_Species, y = lfc)) + theme_bw() + coord_flip() + geom_bar(stat="identity", aes(fill = Genus)) + ggtitle("NEG-ART BOTSWANA") + theme(legend.position = "bottom"), width = 10, height = 10, units = "in", dpi = 300)
ggsave("Figure2BCD_NEG_ART_ANCOM_UGANDA_v7.pdf", ggplot(data = sigtab_dataset_uganda, aes(x = Genus_Species, y = lfc)) + theme_bw() + coord_flip() + geom_bar(stat="identity", aes(fill = Genus)) + ggtitle("NEG-ART UGANDA") + theme(legend.position = "bottom"), width = 10, height = 10, units = "in", dpi = 300)
#Merge results from all three sample cohorts:
sigtab_dataset_art<-rbind(sigtab_dataset_us, sigtab_dataset_botswana, sigtab_dataset_uganda)
sigtab_dataset_art[["index_num_all"]] <- 1:nrow(sigtab_dataset_art)
sigtab_dataset_art <- tidyr::unite(sigtab_dataset_art, col = "Genus_Species_all", Genus, Species, index_num_all, sep = "_", remove = FALSE)
sigtab_dataset_art <- tidyr::unite(sigtab_dataset_art, col = "Genus_Species_all_for_color", Genus, Species, sep = " ", remove = FALSE)
# Remove extra brackets around taxonomic name for labels
sigtab_dataset_art$Genus_label <- stringr::str_replace(sigtab_dataset_art$Genus, "^\\[([^\\]]+)\\]", "\\1")
sigtab_dataset_art$Species_label <- stringr::str_replace(sigtab_dataset_art$Species, "^\\[([^\\]]+)\\]", "\\1")
sigtab_dataset_art %>% dplyr::mutate(Genus_species_label =
dplyr::case_when(is.na(stringr::str_extract(Genus_label, "\\[")) & is.na(stringr::str_extract(Species_label, "\\[")) ~ paste(Genus_label,Species_label),
TRUE ~ paste(Genus_label,"sp"))) -> sigtab_dataset_art
sigtab_dataset_art$Genus_species_label <- stringr::str_replace(sigtab_dataset_art$Genus_species_label, "\\[([^\\]]+)\\]", "")
sigtab_dataset_art$Genus_species_label <- stringr::str_replace(sigtab_dataset_art$Genus_species_label, "\\ +", " ")
sigtab_dataset_art <- dplyr::arrange(sigtab_dataset_art, cohort, lfc)
sigtab_dataset_art$Genus_Species_all<-factor(sigtab_dataset_art$Genus_Species_all, levels = c(sigtab_dataset_art$Genus_Species_all))
# Make lookup table for Genus_species_label
Genus_species_label_lookup <- dplyr::distinct(sigtab_dataset_art, Genus_Species_all, .keep_all = TRUE)$Genus_species_label
names(Genus_species_label_lookup) <- dplyr::distinct(sigtab_dataset_art, Genus_Species_all, .keep_all = TRUE)$Genus_Species_all
#Load color dictionary and construct figure:
dictionary_unique <- read.csv("COLOR_DICTIONARY3.csv", sep=";")
sigtab_dataset_art$Genus_species_label <- factor(sigtab_dataset_art$Genus_species_label, levels = unique(sigtab_dataset_art$Genus_species_label))
dictionary_plot <- dictionary_unique[dictionary_unique$Genus_species_label%in%sigtab_dataset_art$Genus_species_label,,drop=F]
rownames(dictionary_plot)<-dictionary_plot$Genus_species_label
dictionary_plot<-dictionary_plot[as.character(unique(sigtab_dataset_art$Genus_species_label)),,drop=F]
sigtab_dataset_art$cohort <- fct_relevel(as.factor(sigtab_dataset_art$cohort), "boston", "botswana", "uganda_2")
header_namer <- as_labeller(c(`boston` = "U.S.\nHIV-uninfected HIV+ ART-treated", `botswana` = "Botswana\nHIV-uninfected HIV+ ART-treated", `uganda_2` = "Uganda\nHIV-uninfected HIV+ ART-treated"))
ggsave("Figure2BCD_NEG_ART_ANCOM_ALL_v9.pdf",
ggplot(data = sigtab_dataset_art, aes(x = Genus_Species_all, y = lfc)) +
geom_bar(stat = "identity", aes(fill = Genus_species_label)) +
scale_fill_manual(values = as.character(dictionary_plot$Color)) +
coord_flip() + scale_x_discrete(label = as_labeller(Genus_species_label_lookup)) + theme_bw() +
theme(legend.position = "bottom", axis.text.x = element_text(size = 16), axis.text.y = element_text(size = 6, face = "plain"), axis.ticks.y = element_blank(),
panel.border = element_rect(linetype = "solid", fill = NA, linewidth = 1), axis.title.y = element_blank(),
strip.background = element_rect(colour = "white", fill = "white"), strip.text = element_text(colour = "black", face = "bold", size = rel(1.2))) +
geom_hline(yintercept = 0) + ylab("log2FoldChange") +
facet_wrap(vars(cohort), scales = "fixed", ncol = 3, labeller = labeller(cohort = header_namer)), width = 15, height = 20, units = "in", dpi = 300)
#FIGURE 2E
#--------------------------------------------------------------------------------------------------------------
# HEATMAPS
us <- dplyr::filter(.data = sigtab_dataset_art, cohort == "boston")
botswana <- dplyr::filter(.data = sigtab_dataset_art, cohort == "botswana")
uganda <- dplyr::filter(.data = sigtab_dataset_art, cohort == "uganda_2")
length(intersect(us$taxon, botswana$taxon)) -> us_botswana_overlap #3
length(intersect(us$taxon, uganda$taxon)) -> us_uganda_overlap #0
length(intersect(botswana$taxon, uganda$taxon)) -> botswana_uganda_overlap #9
# US-BOTSWANA Overlap
common_us_bots <- c(intersect(us$taxon, botswana$taxon))
botswana_common <- botswana[botswana$taxon%in%common_us_bots,,drop=F]
us_common <- us[us$taxon%in%common_us_bots,,drop=F]
all.equal(botswana_common$taxon, us_common$taxon)## [1] "2 string mismatches"data1 <- cbind(botswana_common$lfc, us_common$lfc)
colnames(data1) <- c("Botswana", "US")
row.names(data1) <- botswana_common$taxon
data1 <- as.data.frame(data1)
data1$Genus_Species_all <- botswana_common$Genus_Species_all
data_melt1 <- melt(data1)## Using Genus_Species_all as id variablesggsave("Figure2E_heat_US_BOTSWANA_v4.pdf",
ggplot(data = data_melt1, aes(x = variable, y = Genus_Species_all)) + geom_tile(aes(fill = value), color = "white", size = 2) +
scale_fill_gradient2(low = "brown", mid = "white", high = "darkblue", limits = c(-1.1,1.1)) + theme_bw() +
scale_y_discrete(label = as_labeller(Genus_species_label_lookup)) +
theme(axis.text.x = element_text(size = 16), axis.text.y = element_text(size = 16, face = "plain"),
axis.ticks = element_blank(), axis.title = element_blank(),
panel.grid = element_line(linewidth = 0.1), panel.background = element_rect(fill = "white")), width = 15, height = 10)
# UGANDA-BOTSWANA Overlap
common_ug_bots <- c(intersect(botswana$taxon, uganda$taxon))
botswana_common <- botswana[botswana$taxon%in%common_ug_bots,,drop=F]
uganda_common <- uganda[uganda$taxon%in%common_ug_bots,,drop=F]
all.equal(botswana_common$taxon, uganda_common$taxon)## [1] "8 string mismatches"data2 <- cbind(botswana_common$lfc, uganda_common$lfc)
colnames(data2) <- c("Botswana", "Uganda")
row.names(data2) <- botswana_common$taxon
data2 <- as.data.frame(data2)
data2$Genus_Species_all <- botswana_common$Genus_Species_all
data_melt2 <- melt(data2)## Using Genus_Species_all as id variablesggsave("Figure2E_heat_UGANDA_BOTSWANA_v2.pdf",
ggplot(data = data_melt2, aes(x = variable, y = Genus_Species_all)) + geom_tile(aes(fill = value), color = "white", size = 2) +
scale_fill_gradient2(low = "brown", mid = "white", high = "darkblue", limits = c(-1.8, 1.8)) + theme_bw() +
scale_y_discrete(label = as_labeller(Genus_species_label_lookup)) +
theme(axis.text.x = element_text(size = 16), axis.text.y = element_text(size = 16, face = "plain"),
axis.ticks = element_blank(), axis.title = element_blank(),
panel.grid = element_line(linewidth = 0.1), panel.background = element_rect(fill = "white")), width = 15, height = 10)
#VENN-EULER DIAGRAMS
BioVenn::draw.venn(us$taxon, botswana$taxon, uganda$taxon, title = NULL, subtitle = NULL,
xtitle = "U.S.", x_c = "royalblue4",
ytitle = "Botswana", y_c = "darkorange",
ztitle = "Uganda", z_c = "forestgreen",
nrtype = "abs",
output = "pdf", filename = "Figure2E_venn_v2.pdf")## [1] "x total: 53"
## [1] "y total: 26"
## [1] "z total: 128"
## [1] "x only: 50"
## [1] "y only: 14"
## [1] "z only: 119"
## [1] "x-y total overlap: 3"
## [1] "x-z total overlap: 0"
## [1] "y-z total overlap: 9"
## [1] "x-y only overlap: 3"
## [1] "x-z only overlap: 0"
## [1] "y-z only overlap: 9"
## [1] "x-y-z overlap: 0"## $x
## [1] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGTCTGAAGTGAAATGCGGGGGCTCAACCCCTGAACTGCTTTGGAAACTGTCAGGCTGGAGTGCAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [2] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCCGTGCAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGTATGGCTGGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [3] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCTGTTTTTTAAGTTAGAGGTGAAAGCTCGACGCTCAACGTCGAAATTGCCTCTGATACTGAGAGACTAGAGTGTAGTTGCGGAAGGCGGAATGTGTGGTGTAGCGGTGAAATGCTTAGATATCACACAGAACACCGATTGCGAAGGCAGCTTTCCAAGCTATTACTGACGCTGAGGCACGAAAGCGTGGGGAGCGAAC"
## [4] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [5] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGTGTAGGCGGGAAGGCAAGTCAGAAGTGAAAATTATGGGCTTAACCCATAACCTGCTTTTGAAACTGTTTTTCTTGAGTGAGGCAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGGCCTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [6] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGTCCTGCAAGTCTGATGTGAAAACCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTAGGACTAGAGTGTCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGACCACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [7] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATGCCAAGTCAGCTGTGAAAACTATGGGCTTAACCTGTAGACTGCAGTTGAAACTGGTATTCTTGAGTGAAGTAGAGGTTGGCGGAATTCCGAGTGTAGCGGTGAAATGCGTAGATATTCGGAGGAACACCGGTGGCGAAGGCGGCCAACTGGGCTTTAACTGACGCTGAGGCTCGAAAGTGTGGGGAGCAAAC"
## [8] "ACAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGCGTAGGCGGACTGTCAAGTCAGTCGTGAAATACCGGGGCTTAACCCCGGGGCTGCGATTGAAACTGACAGCCTTGAGTATCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [9] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGGAAGTGTGGGTAGCAAAC"
## [10] "ACAAGCGTTGTCCGGAACTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [11] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGCACGCCAAGTCAGCGGTGAAATTTTCGGGCTCAACCCGGACTGTGCCGTTGAAACTGGCGAGCTAGAGTGCACAAGAGGCAGGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCCTGCTAGGGTGCGACAGACGCTGAGGCACGAAAGCGTGGGTATCGAAC"
## [12] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGAAGAATAAGTCAGCGGTGAAATGCTTCAGCTCAACTGGAGAATTGCCGATGAAACTGTTTTTCTAGAGTATAAAAGAGGTATGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCATACTGGGCTATAACTGACGCTGAAGCACGAAAGCGTGGGTATCGAAC"
## [13] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCGATCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGATTGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [14] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTTCTTAAGTCCATCTTAAAAGTGCGGGGCTTAACCCCGTGATGGGATGGAAACTGGGAGGCTGGAGTATCGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCGGTGGCGAAGGCGACTTTCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACA"
## [15] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGCATGATAAGTCTGATGTGAAAACCCAAGGCTCAACCATGGGACTGCATTGGAAACTGTCGTGCTGGAGTGTCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [16] "GCGAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGTGTAGGCGGGGTGTCAAGTCAGATGTGAAAACTGTGGGCTCAACCCACAAACTGCATTTGAAACTGATACTCTTGAGAGTGGGAGAGGTAAACGGAATTCCTGGTGTAGTAGTGAAATGCGTAGATATCAGGAGGAACACCGGTGGCGAAGGCGGTTTACTGGACCACAACTGACGCTGAGACACGAAAGCGTGGGGAGCAAAC"
## [17] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGCCAGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTTGGAACTGTCAGGCTAGAGTGTCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [18] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGGAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [19] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGGTGATTAAGTCAGCGGTGAAATGCGTCAGCTTAACTGGCGAACTGCCATTGAAACTGGTTACCTTGAGTGTAGCGGAAGTATGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCATACCATACTATAACTGACGCTGAAGCACGAAAGCGTGGGTATCGAAC"
## [20] "GCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGAAGAATAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAACTGCATCGGAAACTGTTTTTCTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAAC"
## [21] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGTGCGTAGGTGGCAGTGCAAGTCAGATGTGAAAGGCCGGGGCTCAACCCCGGAGCTGCATTTGAAACTGCGCGGCTAGAGTACAGGAGAGGCAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACTGTTACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [22] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGTACGTAGGCGGTTTGCTAAGCGCAAGGTGAAAGGCAGTGGCTTAACCATTGTAAGCCTTGCGAACTGGCAGACTTGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGCTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [23] "GCAAGCGTTGTCCGGAATCACTGGGCGTAAAGGGCGCGTAGGCGGCAATATAAGTCAGATGTGAAAGGTGAGGGCTCAACCCTTAGACTGCATCTGATACTGTATAGCTTGAGTGTGAGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGCTCATAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [24] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGCGATGCAAGTCAGAAGTGAAAGCCCAGGGCTTAACCGTGGGACTGCTTTTGAAACTGTGTTGCTGGATTGCCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTGAATGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [25] "GCGAGCGTTGTCCGGAATTACTGGGCGTAAAGGGAGCGTAGGCGGTCTGATAAGTTGGATGTGAAATACCCGGGCTTAACTTGGGGGGTGCATCCAATACTGTTGGACTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCATAGATATTAGGAGGAACATCGGTGGCGAAGGCGGCTTTCTGGACTGCAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [26] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAAGGTAAGTTAGTTGTGAAATCCCTCGGCTCAACTGAGGAACTGCGACTAAAACTGCTTTTCTTGAGTGCTGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACAGCAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [27] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGTGGACAGTTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGCTGTCTTGAGTACAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [28] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGTGCGTAGGTGGTGAGACAAGTCTGAAGTGAAAATCCGGGGCTTAACCCCGGAACTGCTTTGGAAACTGCCTGACTAGAGTACAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTACTGGACTGCTACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [29] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGTGGCAAGGCAAGCCAGAAGTGAAAACCCGGGGCTCAACCGCGGGATTGCTTTTGGAACTGTCATGCTAGAGTGCAGGAGGGGTGAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [30] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGCCGGCAAGTCAGATGTGAAATCCACGGGCTTAACCCGTGAACTGCATTTGAAACTGTTGGTCTTGAGTATCGGAGAGGTAATCGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGATTACTGGACGACAACTGACGGTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [31] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCGGATTATTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGTAGTCTTGAGTGCAGCAGAGGTAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTACTGGACTGTAACTGACGCTGATGCTCGAAAGTGTGGGTATCAAAC"
## [32] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTTCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
## [33] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGACGGCACTGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGTAGAGCTAGAGTGCTGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [34] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATGCCAAGTCAGCTGTGAAAACTATGGGCTTAACTTGTAGACTGCAGTTGAAACTGGTATTCTTGAGTGAAGTAGAGGTTGGCGGAATTCCGAGTGTAGCGGTGAAATGCGTAGATATTCGGAGGAACACCGGTGGCGAAGGCGGCCAACTGGGCTTTAACTGACGCTGAGGCTCGAAAGTGTGGGGAGCAAAC"
## [35] "GCAAGCGTTATCCGGATCTACTGGGTGTAAAGGGAGCGTAGACGGATGGACAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGCCCGTCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [36] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTATGGCAAGTCTGATGTGAAAGGCCAGGGCTCAACCCTGGGACTGCATTGGAAACTGTCGAACTAGAGTGTCGGAGAGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [37] "GCAAGCGTTGTCCGGAATGATTGGGCGTAAAGGGCGCGTAGGCGGCCAACTAAGTCTGGAGTGAAAGTCCTGCTTTTAAGGTGGGAATTGCTTTGGAAACTGGATGGCTTGAGTGCAGGAGAGGTAAGCGGAATTCCCGGTGTAGCGGTGAAATGCGTAGAGATCGGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGCTGAGGCGCGAAAGTGTGGGGAGCAAAC"
## [38] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGATGGACAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGCCCGTCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATACGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACATTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [39] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGTACGTAGGCGGTCTGGTAAGCGCAAGGTGAAAGGCATAGGCTCAACCAATGTCAGCCTTGCGAACTGTCAGACTTGAGTGCAGGAGGGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACTGTAACTGACGCTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [40] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGACAGCAAGTTGAATGTGAAATCTATGGGCTCAACCCATAAACTGCGTTCAAAACTGTTGTTCTTGAGTGAAGTAGAGGTAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCTTTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCAAAC"
## [41] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGTCCTGCAAGTCTGATGTGAAAGGCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTAGGACTAGAGTGTCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [42] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGGAGACAAGTTGAATGTTTAAACTATCGGCTCAACTGATAGTCGCGTTCAAAACTATCACTCTTGAGTGCAGTAGAGGTAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCAAACA"
## [43] "GCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGTAGGCTGTATGTCAAGTCAAGGGTGAAATCCCACGGCTCAACCGTGGAACTGCCTTTGAAACTGGCAAACTGGAGTATGTGAGAGGGCGGCGGAATTCCTGGTGTAGGAGTGAAATCCGTAGATATCAGGAGGAACATCAGTGGCGAAGGCGGCCGCCTGGCACAAAACTGACGCTGAGGTGCGAAAGCGTGGGTAGCAAAC"
## [44] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGACTGCAAGTCAGGTGTGAAAACCAGGGGCTCAACCTCTGGCCTGCATTTGAAACTGTAGTTCTTGAGTGCTGGAGAGGCAATCGGAATTCCGTGTGTAGCGGTGAAATGCGTAGATATACGGAGGAACACCAGTGGCGAAGGCGGATTGCTGGACAGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [45] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGTGTAGGCGGGACTGCAAGTCAGACGTGAAAATCATGGGCTCAACCCATGACTTGCGTTTGAAACTGCGGTTCTTGAGAGTGGGAGAGGTAAACGGAATTCCTGGTGTAGCGGTGAAATGCGTAGATATCAGGAGGAACACCGGTGGCGAAGGCGGTTTACTGGACCACAACTGACGCTGAGACACGAAAGCGTGGGGAGCAAAC"
## [46] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGATGGATGTTTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGCAGTCTTGAGTGCAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCCTGCTAAGCTGCAACTGACATTGAGGCTCGAAAGTGTGGGTATCAAAC"
## [47] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGCCCTATAAGTCAGCGGTGAAATGTTCCGGCTCAACCGGGAAACTGCCGTTGAAACTGTAGAGCTAGAGTCCACAAGAGGTATGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCATACTGGGGTGAAACAGACGCTGAAGCACGAAAGCGTGGGTATCGAAC"
## [48] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGCGTGTAGGCGGAGCTGCAAGTCAGATGTGAAATCCCGGGGCTCAACCCCGGAACTGCATTTGAAACTGTAGCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [49] "GCGAGCGTTGTCCGGAATTACTGGGCGTAAAGGGCGCGTAGGCGGCCAATTAAGTTAGATGTGAAATCCCCGGGCTTAACCTGGGTGTTGCATTTAAAACTGATAGGCTTGAGTGCAGGAGAGGGAAGCGGAATTCCTAGTGTAGCGGTGGAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGCTGAGGCGCGAGAGCGTGGGGAGCAAAC"
## [50] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGAGTAGGCGGCACGGCAAGTAAGATGTGAAAGCCCGAGGCTTAACCTCGGGATTGCATTTTAAACTGCTGAGCTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGAAACTGACGCTGAGGCACGAAAGCGTGGGGAGCGAAC"
## [51] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGCGTAGGCGGGGATGCAAGTCAGATGTGAAATCTATGGGCTTAACCCATAAACTGCATTTGAAACTGTATCTCTTGAGTGCTGGAGAGGTAGACGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGTCTACTGGACAGTAACTGACGCTGAGGCGCGAGAGCGTGGGGAGCAAAC"
## [52] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGCAGCAAGTCAGAAGTGAAATCTCTGGGCTCAACCCAGAAACTGCTTTTGAAACTGTTGCCCTTGAGTATCGGAGAGGCAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [53] "GCGAGCGTTGTCCGGATTTACTGGGTGTAAAGGGTGCGTAGGCGGCTAGACAAGTCAGGTGTGAAATACCGCAGCTCAACTGCGGGGCTGCACTTGAAACTGTAGAGCTTGAGTGATGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTAAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACATTAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAAC"
##
## $y
## [1] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCGATCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGGTCGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [2] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [3] "GCGAGCGTTGTCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATTTTTAAGTGAGATGTGAAATACTCGGGCTTAACCTGAGTGCTGCATTTCAAACTGGAAGTCTAGAGTGCAGGAGAGGAGAAGGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGCTTCTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [4] "GCGAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTTGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTCCAAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [5] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGCATGCAAGTCAGATGTGAAATCTCAGGGCTTAACCCTGAAACTGCATTTGAAACTGTATGTCTTGAGTGCCGGAGAGGTAATCGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGATTACTGGACGGTAACTGACGGTGAGGCGCGAAAGCGTGGGGAGCGAAC"
## [6] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATGGCAAGTCAGATGTGAAAACTATGGGCTCAACCCATAGACTGCATTTGAAACTGTTGTTCTTGAGTGAGGTAGAGGTAAGCGGAATTCCTGGTGTAGCGGTGAAATGCGTAGAGATCAGGAGGAACATCGGTGGCGAAGGCGGCTTACTGGGCCTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [7] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGTCTGCAAGTCAGATGTGAAATCCATGGGCTCAACCCATGAACTGCATTTGAAACTGTAGATCTTGAGTGTCGGAGGGGCAATCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGATTGCTGGACGATAACTGACGGTGAGGCGCGAAAGTGTGGGGAGCAAAC"
## [8] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGAATGCAAGTCAGATGTGAAATCCATGGGCTTAACCCATGAACTGCATTTGAAACTGTATTTCTTGAGTACTGGAGAGGCAATCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGATTGCTGGACAGCAACTGACGGTGAGGCGCGAAAGTGTGGGGAGCAAAC"
## [9] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTCGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCGAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [10] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTTAGGAGTGAAATCTATGGGCTCAACCCATAAACTGCTTCTAAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [11] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCCGTGCAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGTATGGCTGGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [12] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATTGCAAGTCAGATGTGAAAACTATGGGCTTAACCCATAGACTGCATTTGAAACTGTAGTTCTTGAGTGAAGTAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCGGTGGCGAAGGCGGCTTACTGGGCTTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [13] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGAAGGACAAGTTGGAAGTGAAACCCACGGGCTCAACCCGTGAACTGCTTTCAAAACTGTTTTTCTTGAGTGGTGTAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [14] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGACGGGACTTTAAGTCAGCTGTGAAATTTTCCGGCTCAACCGGGAAACTGCAGTTGATACTGGCGTCCTTGAGTACGGTCGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACCCCGATTGCGAAGGCAGCCTGCCAGACCGCAACTGACGTTCATGCTCGAAAGTGCGGGTATCAAAC"
## [15] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATCTTAAGTCAGGTGTGAAAACTATGGGCTCAACCCATAGACTGCACTTGAAACTGAGGTTCTTGAGTGAAGTAGAGGCAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCCTGCTGGGCTTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [16] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGCGTAGGCGGGGATGCAAGTCAGATGTGAAATCTATGGGCTTAACCCATAAACTGCATTTGAAACTGTATCTCTTGAGTGCTGGAGAGGTAGACGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGTCTACTGGACAGTAACTGACGCTGAGGCGCGAGAGCGTGGGGAGCAAAC"
## [17] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCGGGCTGTTAAGTCAGCGGTCAAATGTCAGGGCCCAACCTTGGCATGCCGTTGATACTGGCGGCCTTGAGTTCACACAAGGAAGGTGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTTCTGGGGTGTTACTGACGCTGAGGCTCGAAAGTGCGGGAATCAAACA"
## [18] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGAAAGCAAGTCAGATGTGAAAACTGTGGGCTCAACCCACAGCCTGCATTTGAAACTGTTTTTCTTGAGTACTGGAGAGGCAGATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGATCTGCTGGACAGCAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [19] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGCGGTATGGCAAGTTTGATGTGAAACCCACAGGCTTAACCTGTGGCTTGCATCGAAAACTACTGAACTAGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGCAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [20] "GCGAGCGTTGTTCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTCGTTAAGTCTGCTGTCAAAGGCTGAGGCTCAACCTCAGTTCTACAACAGATACTGGCGGACTAGAGTATGTGAGAGGGAAGTGGAATTCCCGGTGTAGCGGTGAAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCTTCCTGGCACACTACTGACGCTCATGTGCGAAAGCCAGGGCAGCGAAC"
## [21] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGACGGCACTGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGTAGAGCTAGAGTGCTGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [22] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGAAGCAAGTCTGAAGTGAAAACCCAGGGCTCAACCCTGGGACTGCTTTGGAAACTGTTTTGCTAGAGTGTCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [23] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGCCGGCAAGTCAGATGTGAAATCTGGAGGCTTAACCTCCAAACTGCATTTGAAACTGTAGGTCTTGAGTACCGGAGAGGTTATCGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGATAACTGGACGGCAACTGACGGTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [24] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCGCGCAGGCGGCGTCGTAAGTCGGTCTTAAAAGTGCGGGGCTTAACCCCGTGAGGGGACCGAAACTGCGATGCTAGAGTATCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAAGCGGCTTTCTGGACGACAACTGACGCTGAGGCGCGAAAGCCAGGGGAGCAAACG"
## [25] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGAGCGCAGGCGGGAAACTAAGCGGATCTTAAAAGTGCGGGGCTCAACCCCGTGATGGGGTCCGAACTGGTTTTCTTGAGTGCAGGAGAGGAAAGCGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCTAGGGTAGCGAACG"
## [26] "GCAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGCAGGCGGTTCATCAAGTAGGATGTGAAATCCCCGGGCTCAACCTGGGAACAGCATACTAAACTGGTGGACTAGAGTATTGCAGGGGGAGACGGAATTCCAGGTGTAGCGGTGGAATGCGTAGATATCTGGAAGAACACCAAAGGCGAAGGCAGTCTCCTGGGCAAATACTGACGCTCATATGCGAAAGCGTGGGTAGCAAAC"
##
## $z
## [1] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [2] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [3] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCAGGTTAAGCGTGTTGTGAAATGTAGGGGCTCAACCTCTGCACTGCAGCGCGAACTGGCTTGCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [4] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGAGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [5] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [6] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGGGCGCAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [7] "GCAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGCAGGCGGTTCATCAAGTAGGATGTGAAATCCCCGGGCTCAACCTGGGAACAGCATACTAAACTGGTGGACTAGAGTATTGCAGGGGGAGACGGAATTCCAGGTGTAGCGGTGGAATGCGTAGATATCTGGAAGAACACCAAAGGCGAAGGCAGTCTCCTGGGCAAATACTGACGCTCATATGCGAAAGCGTGGGTAGCAAAC"
## [8] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGAGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [9] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGCAGGCGGCGCGCCAAGTCAGCGGTCAAAGTTCCGGGCTCAACCCGGTGTCGCCGTTGAAACTGGCGTGCTCGAGTGCGTGCGAGGAAGGCGGAATGCGTTGTGTAGCGGTGAAATGCATAGATATGACGCAGAACTCCGATTGCGAAGGCAGCTTTCCAGCGCGCTACTGACGCTGAGGCACGAAAGCGTGGGGATCGAACA"
## [10] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTGCGGGAGTGCGACTGACGCTGAAGCTCGAAGGTGCGGGTATCGAAC"
## [11] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTTCTTAAGTCCATCTTAAAAGTGCGGGGCTTAACCCCGTGATGGGATGGAAACTGAGAGGCTGGAGTATCGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCGGTGGCGAAGGCGACTTTCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACA"
## [12] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGGGCGCAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [13] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTGAGGTAAGCGTGTTGTGAAATGTAGGCGCCCAACGTCTGCACTGCAGCGCGAACTGCCCCACTTGAGTGCGCGCAACGCCGGCGGAACTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACCCCGATTGCGAAGGCAGCTGGCGGGAGCGTAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [14] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGTGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [15] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTTTTAAGCGTGTTGTGAAATACTGTCGCTCAACGACAGAGGTGCAGCGCGAACTGGGAGACTTGAGTGCGCGGAATGCAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCTTGCAGTAGCGTAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [16] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTTATAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGTGAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [17] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [18] "GCGAGCGTTGTTCGGAATCATTGGGCGTAAAGGGCGTGTAGGCGGCCCTGCAAGCCTGGCGTGAAATCCCGGGGCCCAACCCCGGAACCGCGCTGGGAACTGCTGGGCTTGAGCCGCTGTGGCGCAGCCGGAATTCCAGGTGTAGGGGTGAAATCTGTAGATATCTGGAAGAACACCGATGGCGAAGGCAGGCTGCGAGCGGACGGCTGACGCTGAGGCGCGAAGGCGCGGGGAGCGAAC"
## [19] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAAGCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [20] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCAGGTTAAGCGTGTTGTGAAATGTAGGGGCTCAACCTCTGCACTGCAGCGCGAACTGGCTTGCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [21] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCATGTAGGCGGTTTTTTAAGTCTGGAGTGAAAATGCGGGGCTCAACCCCGTATGGCTCTGGATACTGGAAGACTTGAGTGCAGGAGAGGAAAGGGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGCCTTTCTGGACTGTGTCTGACGCTGAGATGCGAAAGCCAGGGTAGCGAACG"
## [22] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTCGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCGAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [23] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [24] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGAACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [25] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGTGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [26] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGCTAAGCGTGCCGTGAAATTCTGTCGCTCAACGGCAGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGACGGTAGCGCAACTGACGCTGAGGCTCGAAAGCGCGGGTATCGAAC"
## [27] "CCAGGCGTTATCCGGATTCATTGGGTTTAAAGGGAGCGTAGGCCGCCTTTTAAGCGTGTTGTGAAATGTAGGGGCTCAACCCCTGCACTGCAGCGCGAACTGGAGGGCTTGAGTACACACAAAGTAGGCGGAATTCATGGTGTAGCGGTGAAATGCTTAGATATCATGAAGAACCCCGATTGCGAAGGCAGCTTACTGGAGTGTAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [28] "GCGAGCGTTGTTCGGAATCATTGGGCGTAAAGGGCATGTAGGCGGTTACGCAAGCCTGGTGTGAAAGCCCGAGGCTCAACCTCGGGATGCGCCGGGAACTGTGCAACTAGAGTAGCTGAGGGGCAGCCGGAATTCCAGGTGTAGGGGTGAAATCTGTAGATATCTGGAAGAACACCGATGGCGAAGGCAGGCTGCCAGCAGATTACTGACGCTGAGGTGCGAAGGTGCGGGGAGCGAACA"
## [29] "GCGAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTTGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTCCAAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [30] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCGGACCTTTAAGTCAGCTGTGAAATACGGCGGCTCAACCGTCGAACTGCAGTTGATACTGGAGGTCTTGAGTGCACACAGGGATACTGGAATTCATGGTGTAGCGGTGAAATGCTCAGATATCATGAAGAACTCCAATCGCGAAGGCAGGTATCCGGGGTGCAACTGACGCTGAGGCTCGAAAGTGCGGGTATCAAAC"
## [31] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGGCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [32] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTGCGGGAGTGCGACTGACGCTGAAGCTCGAAGGTGCGGGTATCGAAC"
## [33] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [34] "GCGAGCGTTGTCCGGAATGACTGGGTGTAAAGGGAGCGTAGGCGGGATGGCAAGTCAGATGTGAAACCTGAGGGCTCAACCTTCAGACTGCATTTGAAACTGCTGTTCTTGAGTGAAGTAGAGGTAAGCGGAATTCCTGGTGTAGCGGTGAAATGCGTAGAGATCAGGAGGAACATCGGTGGCGAAGGCGGCTTACTGGGCTTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [35] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTGCGGGAGTGCGACTGACGCTGAAGCTCGAAGGTGCGGGTATCGAAC"
## [36] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGTTAAGCGTGTCGTGAAATTCCGTCGCTCAACGGCGGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGACGGTAGCGCAACTGACGCTGAGGCTCGAAAGTGCGGGTATCGAAC"
## [37] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTTTGGTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGCCAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [38] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTGGTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGTCAGACTTGAGTGCGCGGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [39] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [40] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGTGTAGGTGGCCATGCAAGTCAGAAGTGAAAATCCGGGGCTCAACCCCGGAACTGCTTTTGAAACTGTAAGGCTGGAGTGCAGGAGGGGTGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTCACTGGACTGTAACTGACACTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [41] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGTTGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [42] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCTGTCCTTTAAGCGTGTTGTGAAATGTAGGCGCTCAACGCCTGCACTGCAGCGCGAACTGAAGGACTTGAGTGTGCACGACGCTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGGCGAGAGCACAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [43] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAGAACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [44] "GCAAGCGTTATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATATTTAAGTGGGATGTGAAATACCCGAGCTTAACTTGGGAGCTGCATTCCAAACTGGATATCTAGAGTGCAGGAGAGGAGAATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGATTCTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [45] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGCTAAGCGTGCCGTGAAATTCTGTCGCTCAACGGCAGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [46] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGCGCAGGCGGGCCGGCAAGTTGGAAGTGAAATCCGGGGGCTTAACCCCCGAACTGCTTTCAAAACTGCTGGTCTTGAGTGATGGAGAGGCAGGCGGAATTCCGTGTGTAGCGGTGAAATGCGTAGATATACGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACATTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [47] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [48] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGACGGTAGCGCAACTGACGCTGAGGCTCGAAAGCGCGGGTATCGAAC"
## [49] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTTATAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGTGAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [50] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTGTTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGGCAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGTAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [51] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGAACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [52] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [53] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGAGTAGGCGGATTGGCAAGTTGGGAGTGAAATGTCGGGGCTTAACCCCGGAACTGCTTCCAAAACTGTTGATCTTGAGTGATGGAGAGGCAGGCGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACATTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCAAAC"
## [54] "GCAAGCGTTAATCGGAATAACTGGGCGTAAAGCGCACGTAGGTGGTTCGACAAGTCAGATGTGAAAGCCCCGGGCTTAACCTGGGATGTGCATTTGAAACTGTTGGACTCGAGTACTGTAGAGGGAGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGATATCTGGAGGAATACCAGTGGCGAAGGCGGCCTCCTGGACAGACACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAAC"
## [55] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCGATCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGGTCGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [56] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCGGGCTGTTAAGTCAGCGGTCAAATGTCAGGGCCCAACCTTGGCATGCCGTTGATACTGGCGGCCTTGAGTTCACACAAGGAAGGTGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTTCTGGGGTGTTACTGACGCTGAGGCTCGAAAGTGCGGGAATCAAACA"
## [57] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGAAGGACAAGTTGGAAGTGAAACCCACGGGCTCAACCCGTGAACTGCTTTCAAAACTGTTTTTCTTGAGTGGTGTAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [58] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCTGTCCTTTAAGCGTGTTGTGAAATGTAGGCGCTCAACGCCTGCACTGCAGCGCGAACTGAAGGACTTGAGTGTGCACGACGCTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [59] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGTTAAGCGTGTCGTGAAATTCCGTCGCTCAACGGCGGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [60] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGAGACAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGCCTTGCTAGAGTGCTGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [61] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGTCGCCGATTAAGTCAGCGGTGAAATCCAGTGGCTCAACCATCGGACTGCCGTTGAAACTGGCCGGCTTGAGTATGATTGAGGCAGGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCTTGCCAAGCCATGACTGACACTGAAGCACGAAAGCGTGGGTATCAAAC"
## [62] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCACAGCAAGTCTGATGTGAAAGCCCGGGGCCCAACCCCGGAACTGCATTGGAAACTGCTGGGCTTGAGTGCAGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [63] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGTCTGATGTGAAAATCCCGGGCTCAACCCGGGAACTGCATTGGAAACTGTTAAGCTAGAGTGCAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [64] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTAAAGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCGCGGGACTGCTTTGGAAACTGTTTAACTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [65] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTGTTAAGCGTGTTGTGAAATGTCGTGGCTCAACCGGGGCACTGCAGCGCGAACTGGCAGACTTGAGTGCACGGTAGGAAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCTTTCCGTAGTGTAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [66] "GCGAGCGTTGTCCGGAATGACTGGGCGTAAAGGGCGTGTAGGCGGCAGTATAAGTCCGGAGTGAAAGTCCTGCTTTCAAGGTGGGAATTGCTTTGGAGACTGTACAGCTTGAGTGCGGAAGAGGTAAGTGGAATTCCCAGTGTAGCGGTGAAATGCGTAGAGATTGGGAGGAACACCAGTGGCGAAGGCGACTTACTGGGCCGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAAC"
## [67] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGCATGCAAGTTGGATGTGAAATGTCACGGCTTAACCGTGGAGCTGCATCCAAAACTGCAAGTCTTGAGTGCCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGCTTTCTGGACGGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [68] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGATGCAAGTCTGAAGTGAAATACCCGGGCTCAACCTGGGAACTGCTTTGGAAACTGTATTGCTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [69] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTTTTTTAAGCGTGTTGTGAAATACTGTCGCTCAACGACAGAGGTGCAGCGCGAACTGGAGGACTTGAGTGCGCGGAATGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCTTACAGTAGCGTAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [70] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGTGTAGGTGGCCAGGCAAGTCAGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTTGAAACTGCAGGGCTAGAGTGCAGGAGGGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACTGTAACTGACACTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [71] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGACGGGACTTTAAGTCAGCTGTGAAATTTTCCGGCTCAACCGGGAAACTGCAGTTGATACTGGCGTCCTTGAGTACGGTCGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACCCCGATTGCGAAGGCAGCCTGCCAGACCGCAACTGACGTTCATGCTCGAAAGTGCGGGTATCAAAC"
## [72] "GCAAGCGTTAATCGGAATCACTGGGCGTAAAGCGCACGTAGGCTGTTATGTAAGTCAGGGGTGAAATCCCACGGCTCAACCGTGGAACTGCCCTTGATACTGCACGACTTGAATCCGGGAGAGGGTGGCGGAATTCCAGGTGTAGGAGTGAAATCCGTAGATATCTGGAGGAACATCAGTGGCGAAGGCGGCCACCTGGACCGGTATTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAAC"
## [73] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTTTGGTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGCCAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [74] "GCAAGCGTTGTCCGGAATCATTGGGCGTAAAGAGTTCGTAGGCGGTTTGTTAAGTCTGGTGTTAAAGCCCGAAGCTCAACTTCGGTTCGGCACTGGATACTGGCAGACTAGAATGCGGTAGAGGTAAAGGGAATTCCTGGTGTAGCGGTGAAATGCGTAGATATCAGGAGGAACATCGGTGGCGTAAGCGCTTTACTGGGCCGTAATTGACGCTGAGGAACGAAAGCCAGGGTAGCGAAT"
## [75] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTTAAGCAAGTCAGAAGTGAAAGGCTGGGGCTCAACCCCGGGACTGCTTTTGAAACTGTTTAACTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [76] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGAAAGCAAGTCAGATGTGAAAACTGTGGGCTCAACCCACAGCCTGCATTTGAAACTGTTTTTCTTGAGTACTGGAGAGGCAGATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGATCTGCTGGACAGCAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [77] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCATGCAAGTTGGAAGTGAAAACTATGGGCTCAACCCATAGCCTGCTTTCAAAACTGCGTGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [78] "GCGAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGTGTAGGCGGGAAGGCAAGTCAGATGTGAAAACCAAAGGCTCAACCTTTGGCTTGCATTTGAAACTGTTTTTCTTGAGAGTGGGAGAGGTAAACGGAATTCCTAGTGTAGTAGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGTTTACTGGACCACAACTGACGCTGAGACACGAAAGCGTGGGGAGCAAAC"
## [79] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGCATGGTAAGTAAGATGTGAAAGCCCGAGGCTTAACCTCGAGGATTGCATTTTAAACTATCAAGCTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGAAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCGAA"
## [80] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTCAAGCAAGTCAGAAGTGAAAGGCTGGGGCTCAACCCCGGGACTGCTTTTGAAACTGTTTGACTGGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [81] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGACGCAAGTCTGAAGTGAAATACCCGGGCTCAACCTGGGAACTGCTTTGGAAACTGTGTTGCTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [82] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGGAGAGCAAGTCAGCGGTGAAATACATGGGCTTAACCCATGGGCTGCCGTTGAAACTGTCCTTCTTGAGTGAAGTAGAGGCAAGCGGAATTCCGAGTGTAGCGGTGAAATGCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGGCTTTTACTGACGCTGAGGCTCGAAAGTGTGGGGAGCAAAC"
## [83] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGTCTGGCAAGTCTGATGTGAAATCCCGGGGCTCAACCCTGGAACTGCATTGGAAACTGTCAGACTAGAGTGCCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [84] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTAAGCAAGTCAGAAGTGAAAGGCTGGGGCTCAACCCCGGGACTGCTTTTGAAACTGTTTGGCTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [85] "GCAAGCGTTACTCGGAATTACTAGGCGTAAAGCGCGCGTAGGCGGAATGTTAAGTCTGTTGTGTAATCTCTGGGCTCAACCCAGAAACTGCAACAGAAACTGGCGTTCTTGAGTGAGGCAGAGGAAATCGGAATTCCTAGTGTAGCAGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGATTTCTGGGCCTTTACTGACGCTAAAGTGCGAAAGCTAGGGGAGCAAAC"
## [86] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGATGCCTAAGCGGGACCTCTAACCCGGGGGCTCAACCCCCGGCCGGGTCCCGAACTGGGCGTCTCGAGTGCGGTAGGGGCAGGTGGAATTCCATGTGTAGCGGTGGAATGCGCAGATATATGGAAGAACACCGACGGCGAAGGCAGCCTGCTGGGCCGACACTGACGCTGAGGTGCGAAAGCGCGGGGAGCGAACA"
## [87] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTAGTGCAAGTCTGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTATAACTAGAGTGTCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [88] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGAAGATAAGTTGGACGTCTAATCTATCGGCTCAACCGATAGTCGCGTTCAAAACTGTTTTTCTTGAGTGAAGTAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAAATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGGCTTTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCAAACA"
## [89] "GCGAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAAATTAAGTTAGTTGTGAAATCCCTCGGCTCAACTGAGGAACTGCAACTAAAACTGATTTTCTTGAGTACTGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACAGAAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [90] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTTGTAGGCGGTTTGTCGCGTCTGCTGTGAAAGGCCGGGGCTTAACCCCGTGTATTGCAGTGGGTACGGGCAGACTAGAGTGCAGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGGAATGCGCAGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCTCTGGGCTGTAACTGACGCTGAGAAGCGAAAGCATGGGGAGCGAA"
## [91] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAATACAAGTTAGTTGTGAAATCCCTCGGCTTAACTGAGGAACTGCAACTAAAACTATATTTCTTGAGTGCTGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACAGTAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [92] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGTGGATTGTTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGAAACTGGCAGTCTTGAGTACAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTAGACTGCAACTGACACTGATGCTCGAAAGTGTGGGTATCAAAC"
## [93] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGTATGGCAAGTCTGATGTGAAAGGCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTCACACTTGAGTGTCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGACAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [94] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAAATTAAGTTAGTTGTGAAATCCCTCGGCTCAACTGAGGAACTGCAACTAAAACTGGTTTTCTTGAGTGCAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTACTGGACTGTAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [95] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAAACCATGGGCTCAACCCATGAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGTCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [96] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTGTGTAAGTCTGAAGTGAAAGCCCGGGGCTCAACCGCGGGACTGCTTTGGAAACTATGCAGCTAGAGTGTCGGAGAGGTAAGTGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [97] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCGCTCGAGCGGGACCTCTAACCCGGGGGCTCAACCTCCGGCCGGGTCCCGGACCGTGCGGCTCGGGTGCGGTAGGGGCAGGCGGAACTCCAAGTGTAGCGGTGAAATGCGCAGATATTTGGAGGAACACCGATGGCGAAGGCAGCCTGCTGGGCCGCCACCGACGCTGAGGCGCGAAAGCCGGGGGAGCGAACA"
## [98] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGTGGTATGGCAAGTCAGAAGTGAAAACCCAGGGCTTAACTCTGGGACTGCTTTTGAAACTGTCAGACTAGAGTGCAGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCTTACTGGACTGAAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [99] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTGTGGCAAGTCTGATGTGAAAGGCATGGGCTCAACCTGTGGACTGCATTGGAAACTGTCATACTTGAGTGCCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [100] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCGACGGCGAAGGCAGCTCTCTGGGCCGAAACTGACGCTGAGGCGCGAAAGCTGGGGGAGCGAAC"
## [101] "CCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGACGGTTATTTAAGTCTGAAGTGAAAGCCCTCAGCTCAACTGAGGAATTGCTTTGGAAACTGGATGACTTGAGTGCAGTAGAGGAAAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGTTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [102] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGTACGTAGGTGGTTACCTAAGCACGAGGTATAAGGCAATGGCTTAACCATTGTTCGCCTTGTGAACTGGGCTACTTGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACACTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [103] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCACGGCAAGCCAGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTTGGAACTGCTGAGCTAGAGTGTCGGAGAGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [104] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGTACGTAGGTGGTTTTCTAAGCACGGGGTTTAAGGCAATGGCTTAACCATTGTTCGCCTTGTGAACTGGAAGACTTGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACACTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [105] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTTTGCAAGTCTGATGTGAAAGGCGGGGGCTCAACCCCTGGACTGCATTGGAAACTGTGAGGCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [106] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGACCGGCAAGTTGGAAGTGAAAACTATGGGCTCAACCCATAAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGACCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [107] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGACCGGCAAGTTGGAAGTGAAAACCATGGGCTCAACCCGTGAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGACCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [108] "GCGAGCGTTATCCGGAATCATTGGGCGTAAAGCGCGCGCAGGCGGGCTTTCAAGCGGCGGCGTCGAAGCCGGGGGCTCAACCCCCGGAAGCGCCCCGAACTGGAAGCCTCGGATGCGGCAGGGGGAGGCGGAATTCCCGGTGTAGCGGTGAAATGCGCAGATATCGGGAAGAACACCGACGGCGAAGGCAGCCTCCTGGGCCGGCATCGACGCTGAGGCGCGAAAGCTGGGGGAGCGAAC"
## [109] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGAGCGCTAAGCGGGACCTCTAACCCGAGGGCTCAACCCCCGGCCGGGTCCCGAACTGGCGCTCTCGAGTGCGGTAGGGGAGAGCGGAATTCCCGGTGTAGCGGTGGAATGCGCAGATATCGGGAAGAACACCGACGGCGAAGGCAGCTCTCTGGGCCGAAACTGACGCTGAGGCGCGAAAGCTGGGGGAGCGAACA"
## [110] "GCTAGCGTTATCCGGATTTACTGGGCGTAAAGGGTGCGTAGGCGGTCTTTTAAGTCAGGAGTGAAAGGCTACGGCTCAACCGTAGTAAGCTCTTGAAACTGGAGGACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTAGCGAAGGCGGCTCTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAACA"
## [111] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGTGGTATGGCAAGTCAGAAGTGAAAACCCAGGGCTTAACTCTGGGACTGCTTTTGAAACTGTCAGACTGGAGTGCAGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCTTACTGGACTGAAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [112] "GCGAGCGTTATCCGGAATGATTGGGCGTAAAGCGCGCGCAGGCGGCCGCTCAAGCGGGACCTCTAACCCCGGGGCTCAACCCCGGGCCGGGTCCCGAACTGGGCGGCTCGAGTGCGGTAGGGGAGAGCGGAATTCCAAGTGTAGCGGTGAAATGCGCAGATATTTGGAAGAACACCGATGGCGAAGGCAGCTCTCTGGGCCGTCACTGACGCTGAGGCGCGAAAGCCGGGGGAGCGAACA"
## [113] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTTGTAGGCGGTTGGTCGCGTCTGCCGTGAAATCCTCTGGCTTAACTGGGGGCGTGCGGTGGGTACGGGCTGACTTGAGTGCGGTAGGGGAGACTGGAACTCCTGGTGTAGCGGTGGAATGCGCAGATATCAGGAAGAACACCGGTGGCGAAGGCGGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
## [114] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGAATGGCAAGTCTGATGTGAAAGGCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTCAATCTAGAGTACCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [115] "TCTAGTGGTAGCAGTTTTTATTGGGCCTAAAGCGTCCGTAGCCGGTTTAATAAGTCTCTGGTGAAATCCTGCAGCTTAACTGTGGGAATTGCTGGAGATACTATTAGACTTGAGATCGGGAGAGGTTAGAGGTACTCCCAGGGTAGAGGTGAAATTCTGTAATCCTGGGAGGACCGCCTGTTGCGAAGGCGTCTGACTGGAACGATTCTGACGGTGAGGGACGAAAGCTAGGGGCGCGAA"
## [116] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTATGGCAAGTCTGATGTGAAAGGCCAGGGCTCAACCCTGGGACTGCATTGGAAACTGTCGAACTAGAGTGTCGGAGAGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [117] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGAGCGCTAAGCGGGACCTCTAACCCGAGGGCTCAACCCCCGGCCGGGTCCCGAACTGGCGCTCTCGAGTGCGGTAGGGGAGAGCGGAATTCCCGGTGTAGCGGTGGAATGCGCAGATATCGGGAGGAACACCGACGGCGAAGGCAGCTCTCTGGGCCGAAACTGACGCTGAGGCGCGAAAGCTGGGGGAGCGAACA"
## [118] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTGTGCAAGTCTGAAGTGAAAGGCATGGGCTCAACCTGTGGACTGCTTTGGAAACTGTGCAGCTAGAGTGTCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [119] "GCTAGCGTTATCCGGAATTACTGGGCGTAAAGGGTGCGTAGGTGGTTTCTTAAGTCAGAGGTGAAAGGCTACGGCTCAACCGTAGTAAGCCTTTGAAACTGGGAAACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTTGCGAAGGCGGCTCTCTGGACTGTAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAACA"
## [120] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGTCAAGCGGGGTTTCAAATCCAGGGGCTCAACCTCTGGCCGGACCCCGAACTGGCGGGCTCGAGTGCGGTAGAGGAAGGTGGAATTCCCAGTGTAGCGGTGAAATGCGCAGATATTGGGAAGAACACCGATGGCGAAGGCAGCCTTCTGGGCCGCCACTGACGCTGAGGCGCGAAAGCTAGGGGAGCGAACA"
## [121] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGCAGCAAGTCTGATGTGAAAGGCAGGGGCTTAACCCCTGGACTGCATTGGAAACTGCTGTGCTTGAGTGCCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [122] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGATTAGCAAGTCTGATGTGAAAGGCAGGGGCTCAACCCCTGGACTGCATTGGAAACTGCCAGTCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGCAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [123] "TCAAGCGTTGTTCGGAATCACTGGGCGTAAAGCGTGCGTAGGCTGTTTCGTAAGTCGTGTGTGAAAGGCGCGGGCTCAACCCGCGGACGGCACATGATACTGCGAGACTAGAGTAATGGAGGGGGAACCGGAATTCTCGGTGTAGCAGTGAAATGCGTAGATATCGAGAGGAACACTCGTGGCGAAGGCGGGTTCCTGGACATTAACTGACGCTGAGGCACGAAGGCCAGGGGAGCGAAA"
## [124] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGGCAGGCCGGGGGTCGAAGCGGGGGGCTCAACCCCCCGAAGCCCCCGGAACCTCCGCGGCTTGGGTCCGGTAGGGGAGGGTGGAACACCCGGTGTAGCGGTGGAATGCGCAGATATCGGGTGGAACACCGGTGGCGAAGGCGGCCCTCTGGGCCGAGACCGACGCTGAGGCGCGAAAGCTGGGGGAGCGAAC"
## [125] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGACCGGCAAGTTGGAAGTGAAATCCATGGGCTCAACCCGTGAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGACCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [126] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAAACCATGGGCTCAACCCATGAATTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGATGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGTCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [127] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
## [128] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
##
## $x_only
## [1] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGTCTGAAGTGAAATGCGGGGGCTCAACCCCTGAACTGCTTTGGAAACTGTCAGGCTGGAGTGCAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [2] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCTGTTTTTTAAGTTAGAGGTGAAAGCTCGACGCTCAACGTCGAAATTGCCTCTGATACTGAGAGACTAGAGTGTAGTTGCGGAAGGCGGAATGTGTGGTGTAGCGGTGAAATGCTTAGATATCACACAGAACACCGATTGCGAAGGCAGCTTTCCAAGCTATTACTGACGCTGAGGCACGAAAGCGTGGGGAGCGAAC"
## [3] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [4] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGTGTAGGCGGGAAGGCAAGTCAGAAGTGAAAATTATGGGCTTAACCCATAACCTGCTTTTGAAACTGTTTTTCTTGAGTGAGGCAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGGCCTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [5] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGTCCTGCAAGTCTGATGTGAAAACCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTAGGACTAGAGTGTCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGACCACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [6] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATGCCAAGTCAGCTGTGAAAACTATGGGCTTAACCTGTAGACTGCAGTTGAAACTGGTATTCTTGAGTGAAGTAGAGGTTGGCGGAATTCCGAGTGTAGCGGTGAAATGCGTAGATATTCGGAGGAACACCGGTGGCGAAGGCGGCCAACTGGGCTTTAACTGACGCTGAGGCTCGAAAGTGTGGGGAGCAAAC"
## [7] "ACAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGCGTAGGCGGACTGTCAAGTCAGTCGTGAAATACCGGGGCTTAACCCCGGGGCTGCGATTGAAACTGACAGCCTTGAGTATCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [8] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGGAAGTGTGGGTAGCAAAC"
## [9] "ACAAGCGTTGTCCGGAACTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [10] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGCACGCCAAGTCAGCGGTGAAATTTTCGGGCTCAACCCGGACTGTGCCGTTGAAACTGGCGAGCTAGAGTGCACAAGAGGCAGGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCCTGCTAGGGTGCGACAGACGCTGAGGCACGAAAGCGTGGGTATCGAAC"
## [11] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGAAGAATAAGTCAGCGGTGAAATGCTTCAGCTCAACTGGAGAATTGCCGATGAAACTGTTTTTCTAGAGTATAAAAGAGGTATGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCATACTGGGCTATAACTGACGCTGAAGCACGAAAGCGTGGGTATCGAAC"
## [12] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCGATCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGATTGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [13] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTTCTTAAGTCCATCTTAAAAGTGCGGGGCTTAACCCCGTGATGGGATGGAAACTGGGAGGCTGGAGTATCGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCGGTGGCGAAGGCGACTTTCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACA"
## [14] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGCATGATAAGTCTGATGTGAAAACCCAAGGCTCAACCATGGGACTGCATTGGAAACTGTCGTGCTGGAGTGTCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [15] "GCGAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGTGTAGGCGGGGTGTCAAGTCAGATGTGAAAACTGTGGGCTCAACCCACAAACTGCATTTGAAACTGATACTCTTGAGAGTGGGAGAGGTAAACGGAATTCCTGGTGTAGTAGTGAAATGCGTAGATATCAGGAGGAACACCGGTGGCGAAGGCGGTTTACTGGACCACAACTGACGCTGAGACACGAAAGCGTGGGGAGCAAAC"
## [16] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGCCAGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTTGGAACTGTCAGGCTAGAGTGTCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [17] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGGAGTGAAATCTATGGGCTCAACCCATAAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [18] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGGTGATTAAGTCAGCGGTGAAATGCGTCAGCTTAACTGGCGAACTGCCATTGAAACTGGTTACCTTGAGTGTAGCGGAAGTATGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACTCCGATTGCGAAGGCAGCATACCATACTATAACTGACGCTGAAGCACGAAAGCGTGGGTATCGAAC"
## [19] "GCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGAAGAATAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAACTGCATCGGAAACTGTTTTTCTTGAGTGCAGAAGAGGAGAGTGGAACTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAAC"
## [20] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGTGCGTAGGTGGCAGTGCAAGTCAGATGTGAAAGGCCGGGGCTCAACCCCGGAGCTGCATTTGAAACTGCGCGGCTAGAGTACAGGAGAGGCAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACTGTTACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [21] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGTACGTAGGCGGTTTGCTAAGCGCAAGGTGAAAGGCAGTGGCTTAACCATTGTAAGCCTTGCGAACTGGCAGACTTGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGCTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [22] "GCAAGCGTTGTCCGGAATCACTGGGCGTAAAGGGCGCGTAGGCGGCAATATAAGTCAGATGTGAAAGGTGAGGGCTCAACCCTTAGACTGCATCTGATACTGTATAGCTTGAGTGTGAGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGCTCATAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [23] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGCGATGCAAGTCAGAAGTGAAAGCCCAGGGCTTAACCGTGGGACTGCTTTTGAAACTGTGTTGCTGGATTGCCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTGAATGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [24] "GCGAGCGTTGTCCGGAATTACTGGGCGTAAAGGGAGCGTAGGCGGTCTGATAAGTTGGATGTGAAATACCCGGGCTTAACTTGGGGGGTGCATCCAATACTGTTGGACTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCATAGATATTAGGAGGAACATCGGTGGCGAAGGCGGCTTTCTGGACTGCAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [25] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAAGGTAAGTTAGTTGTGAAATCCCTCGGCTCAACTGAGGAACTGCGACTAAAACTGCTTTTCTTGAGTGCTGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACAGCAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [26] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGTGGACAGTTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGCTGTCTTGAGTACAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [27] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGTGCGTAGGTGGTGAGACAAGTCTGAAGTGAAAATCCGGGGCTTAACCCCGGAACTGCTTTGGAAACTGCCTGACTAGAGTACAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTACTGGACTGCTACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [28] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGTGGCAAGGCAAGCCAGAAGTGAAAACCCGGGGCTCAACCGCGGGATTGCTTTTGGAACTGTCATGCTAGAGTGCAGGAGGGGTGAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [29] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGCCGGCAAGTCAGATGTGAAATCCACGGGCTTAACCCGTGAACTGCATTTGAAACTGTTGGTCTTGAGTATCGGAGAGGTAATCGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGATTACTGGACGACAACTGACGGTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [30] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCGGATTATTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGTAGTCTTGAGTGCAGCAGAGGTAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTACTGGACTGTAACTGACGCTGATGCTCGAAAGTGTGGGTATCAAAC"
## [31] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTTCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
## [32] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATGCCAAGTCAGCTGTGAAAACTATGGGCTTAACTTGTAGACTGCAGTTGAAACTGGTATTCTTGAGTGAAGTAGAGGTTGGCGGAATTCCGAGTGTAGCGGTGAAATGCGTAGATATTCGGAGGAACACCGGTGGCGAAGGCGGCCAACTGGGCTTTAACTGACGCTGAGGCTCGAAAGTGTGGGGAGCAAAC"
## [33] "GCAAGCGTTATCCGGATCTACTGGGTGTAAAGGGAGCGTAGACGGATGGACAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGCCCGTCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [34] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTATGGCAAGTCTGATGTGAAAGGCCAGGGCTCAACCCTGGGACTGCATTGGAAACTGTCGAACTAGAGTGTCGGAGAGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [35] "GCAAGCGTTGTCCGGAATGATTGGGCGTAAAGGGCGCGTAGGCGGCCAACTAAGTCTGGAGTGAAAGTCCTGCTTTTAAGGTGGGAATTGCTTTGGAAACTGGATGGCTTGAGTGCAGGAGAGGTAAGCGGAATTCCCGGTGTAGCGGTGAAATGCGTAGAGATCGGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGCTGAGGCGCGAAAGTGTGGGGAGCAAAC"
## [36] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGATGGACAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGCCCGTCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATACGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACATTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [37] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGTACGTAGGCGGTCTGGTAAGCGCAAGGTGAAAGGCATAGGCTCAACCAATGTCAGCCTTGCGAACTGTCAGACTTGAGTGCAGGAGGGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACTGTAACTGACGCTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [38] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGACAGCAAGTTGAATGTGAAATCTATGGGCTCAACCCATAAACTGCGTTCAAAACTGTTGTTCTTGAGTGAAGTAGAGGTAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCTTTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCAAAC"
## [39] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGTCCTGCAAGTCTGATGTGAAAGGCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTAGGACTAGAGTGTCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [40] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGGAGACAAGTTGAATGTTTAAACTATCGGCTCAACTGATAGTCGCGTTCAAAACTATCACTCTTGAGTGCAGTAGAGGTAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCTGTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCAAACA"
## [41] "GCGAGCGTTAATCGGAATTACTGGGCGTAAAGCGCACGTAGGCTGTATGTCAAGTCAAGGGTGAAATCCCACGGCTCAACCGTGGAACTGCCTTTGAAACTGGCAAACTGGAGTATGTGAGAGGGCGGCGGAATTCCTGGTGTAGGAGTGAAATCCGTAGATATCAGGAGGAACATCAGTGGCGAAGGCGGCCGCCTGGCACAAAACTGACGCTGAGGTGCGAAAGCGTGGGTAGCAAAC"
## [42] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGACTGCAAGTCAGGTGTGAAAACCAGGGGCTCAACCTCTGGCCTGCATTTGAAACTGTAGTTCTTGAGTGCTGGAGAGGCAATCGGAATTCCGTGTGTAGCGGTGAAATGCGTAGATATACGGAGGAACACCAGTGGCGAAGGCGGATTGCTGGACAGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [43] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGTGTAGGCGGGACTGCAAGTCAGACGTGAAAATCATGGGCTCAACCCATGACTTGCGTTTGAAACTGCGGTTCTTGAGAGTGGGAGAGGTAAACGGAATTCCTGGTGTAGCGGTGAAATGCGTAGATATCAGGAGGAACACCGGTGGCGAAGGCGGTTTACTGGACCACAACTGACGCTGAGACACGAAAGCGTGGGGAGCAAAC"
## [44] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGATGGATGTTTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGATACTGGCAGTCTTGAGTGCAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCCTGCTAAGCTGCAACTGACATTGAGGCTCGAAAGTGTGGGTATCAAAC"
## [45] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGCGGCCCTATAAGTCAGCGGTGAAATGTTCCGGCTCAACCGGGAAACTGCCGTTGAAACTGTAGAGCTAGAGTCCACAAGAGGTATGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCATACTGGGGTGAAACAGACGCTGAAGCACGAAAGCGTGGGTATCGAAC"
## [46] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGCGTGTAGGCGGAGCTGCAAGTCAGATGTGAAATCCCGGGGCTCAACCCCGGAACTGCATTTGAAACTGTAGCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [47] "GCGAGCGTTGTCCGGAATTACTGGGCGTAAAGGGCGCGTAGGCGGCCAATTAAGTTAGATGTGAAATCCCCGGGCTTAACCTGGGTGTTGCATTTAAAACTGATAGGCTTGAGTGCAGGAGAGGGAAGCGGAATTCCTAGTGTAGCGGTGGAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGCTGAGGCGCGAGAGCGTGGGGAGCAAAC"
## [48] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGAGTAGGCGGCACGGCAAGTAAGATGTGAAAGCCCGAGGCTTAACCTCGGGATTGCATTTTAAACTGCTGAGCTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGAAACTGACGCTGAGGCACGAAAGCGTGGGGAGCGAAC"
## [49] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGCAGCAAGTCAGAAGTGAAATCTCTGGGCTCAACCCAGAAACTGCTTTTGAAACTGTTGCCCTTGAGTATCGGAGAGGCAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [50] "GCGAGCGTTGTCCGGATTTACTGGGTGTAAAGGGTGCGTAGGCGGCTAGACAAGTCAGGTGTGAAATACCGCAGCTCAACTGCGGGGCTGCACTTGAAACTGTAGAGCTTGAGTGATGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTAAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACATTAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAAC"
##
## $y_only
## [1] "GCGAGCGTTGTCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATTTTTAAGTGAGATGTGAAATACTCGGGCTTAACCTGAGTGCTGCATTTCAAACTGGAAGTCTAGAGTGCAGGAGAGGAGAAGGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGCTTCTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [2] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGCATGCAAGTCAGATGTGAAATCTCAGGGCTTAACCCTGAAACTGCATTTGAAACTGTATGTCTTGAGTGCCGGAGAGGTAATCGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGATTACTGGACGGTAACTGACGGTGAGGCGCGAAAGCGTGGGGAGCGAAC"
## [3] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATGGCAAGTCAGATGTGAAAACTATGGGCTCAACCCATAGACTGCATTTGAAACTGTTGTTCTTGAGTGAGGTAGAGGTAAGCGGAATTCCTGGTGTAGCGGTGAAATGCGTAGAGATCAGGAGGAACATCGGTGGCGAAGGCGGCTTACTGGGCCTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [4] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGTCTGCAAGTCAGATGTGAAATCCATGGGCTCAACCCATGAACTGCATTTGAAACTGTAGATCTTGAGTGTCGGAGGGGCAATCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGATTGCTGGACGATAACTGACGGTGAGGCGCGAAAGTGTGGGGAGCAAAC"
## [5] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGAATGCAAGTCAGATGTGAAATCCATGGGCTTAACCCATGAACTGCATTTGAAACTGTATTTCTTGAGTACTGGAGAGGCAATCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGATTGCTGGACAGCAACTGACGGTGAGGCGCGAAAGTGTGGGGAGCAAAC"
## [6] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTTAGGAGTGAAATCTATGGGCTCAACCCATAAACTGCTTCTAAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [7] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATTGCAAGTCAGATGTGAAAACTATGGGCTTAACCCATAGACTGCATTTGAAACTGTAGTTCTTGAGTGAAGTAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCGGTGGCGAAGGCGGCTTACTGGGCTTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [8] "GCGAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGATCTTAAGTCAGGTGTGAAAACTATGGGCTCAACCCATAGACTGCACTTGAAACTGAGGTTCTTGAGTGAAGTAGAGGCAGGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCCTGCTGGGCTTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [9] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGCGGTATGGCAAGTTTGATGTGAAACCCACAGGCTTAACCTGTGGCTTGCATCGAAAACTACTGAACTAGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGCAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [10] "GCGAGCGTTGTTCGGAATTACTGGGCGTAAAGCGCACGCAGGCGGTTCGTTAAGTCTGCTGTCAAAGGCTGAGGCTCAACCTCAGTTCTACAACAGATACTGGCGGACTAGAGTATGTGAGAGGGAAGTGGAATTCCCGGTGTAGCGGTGAAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCTTCCTGGCACACTACTGACGCTCATGTGCGAAAGCCAGGGCAGCGAAC"
## [11] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGAAGCAAGTCTGAAGTGAAAACCCAGGGCTCAACCCTGGGACTGCTTTGGAAACTGTTTTGCTAGAGTGTCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [12] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGCAGCCGGGCCGGCAAGTCAGATGTGAAATCTGGAGGCTTAACCTCCAAACTGCATTTGAAACTGTAGGTCTTGAGTACCGGAGAGGTTATCGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGATAACTGGACGGCAACTGACGGTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [13] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCGCGCAGGCGGCGTCGTAAGTCGGTCTTAAAAGTGCGGGGCTTAACCCCGTGAGGGGACCGAAACTGCGATGCTAGAGTATCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAAGCGGCTTTCTGGACGACAACTGACGCTGAGGCGCGAAAGCCAGGGGAGCAAACG"
## [14] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGGGAGCGCAGGCGGGAAACTAAGCGGATCTTAAAAGTGCGGGGCTCAACCCCGTGATGGGGTCCGAACTGGTTTTCTTGAGTGCAGGAGAGGAAAGCGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCTAGGGTAGCGAACG"
##
## $z_only
## [1] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [2] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [3] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCAGGTTAAGCGTGTTGTGAAATGTAGGGGCTCAACCTCTGCACTGCAGCGCGAACTGGCTTGCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [4] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGAGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [5] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [6] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGGGCGCAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [7] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGAGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [8] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGCAGGCGGCGCGCCAAGTCAGCGGTCAAAGTTCCGGGCTCAACCCGGTGTCGCCGTTGAAACTGGCGTGCTCGAGTGCGTGCGAGGAAGGCGGAATGCGTTGTGTAGCGGTGAAATGCATAGATATGACGCAGAACTCCGATTGCGAAGGCAGCTTTCCAGCGCGCTACTGACGCTGAGGCACGAAAGCGTGGGGATCGAACA"
## [9] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTGCGGGAGTGCGACTGACGCTGAAGCTCGAAGGTGCGGGTATCGAAC"
## [10] "GCAAGCGTTGTCCGGAATTATTGGGCGTAAAGCGCGCGCAGGCGGCTTCTTAAGTCCATCTTAAAAGTGCGGGGCTTAACCCCGTGATGGGATGGAAACTGAGAGGCTGGAGTATCGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCGGTGGCGAAGGCGACTTTCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAACA"
## [11] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGGGCGCAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [12] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTGAGGTAAGCGTGTTGTGAAATGTAGGCGCCCAACGTCTGCACTGCAGCGCGAACTGCCCCACTTGAGTGCGCGCAACGCCGGCGGAACTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACCCCGATTGCGAAGGCAGCTGGCGGGAGCGTAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [13] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGTGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [14] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTTTTAAGCGTGTTGTGAAATACTGTCGCTCAACGACAGAGGTGCAGCGCGAACTGGGAGACTTGAGTGCGCGGAATGCAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCTTGCAGTAGCGTAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [15] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTTATAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGTGAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [16] "GCGAGCGTTGTTCGGAATCATTGGGCGTAAAGGGCGTGTAGGCGGCCCTGCAAGCCTGGCGTGAAATCCCGGGGCCCAACCCCGGAACCGCGCTGGGAACTGCTGGGCTTGAGCCGCTGTGGCGCAGCCGGAATTCCAGGTGTAGGGGTGAAATCTGTAGATATCTGGAAGAACACCGATGGCGAAGGCAGGCTGCGAGCGGACGGCTGACGCTGAGGCGCGAAGGCGCGGGGAGCGAAC"
## [17] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAAGCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [18] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCAGGTTAAGCGTGTTGTGAAATGTAGGGGCTCAACCTCTGCACTGCAGCGCGAACTGGCTTGCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [19] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCATGTAGGCGGTTTTTTAAGTCTGGAGTGAAAATGCGGGGCTCAACCCCGTATGGCTCTGGATACTGGAAGACTTGAGTGCAGGAGAGGAAAGGGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGCCTTTCTGGACTGTGTCTGACGCTGAGATGCGAAAGCCAGGGTAGCGAACG"
## [20] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [21] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGAACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [22] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGTGGCTCAACCTCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [23] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGCTAAGCGTGCCGTGAAATTCTGTCGCTCAACGGCAGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGACGGTAGCGCAACTGACGCTGAGGCTCGAAAGCGCGGGTATCGAAC"
## [24] "CCAGGCGTTATCCGGATTCATTGGGTTTAAAGGGAGCGTAGGCCGCCTTTTAAGCGTGTTGTGAAATGTAGGGGCTCAACCCCTGCACTGCAGCGCGAACTGGAGGGCTTGAGTACACACAAAGTAGGCGGAATTCATGGTGTAGCGGTGAAATGCTTAGATATCATGAAGAACCCCGATTGCGAAGGCAGCTTACTGGAGTGTAACTGACGCTGAAGCTCGAAAGCGCGGGTATCGAAC"
## [25] "GCGAGCGTTGTTCGGAATCATTGGGCGTAAAGGGCATGTAGGCGGTTACGCAAGCCTGGTGTGAAAGCCCGAGGCTCAACCTCGGGATGCGCCGGGAACTGTGCAACTAGAGTAGCTGAGGGGCAGCCGGAATTCCAGGTGTAGGGGTGAAATCTGTAGATATCTGGAAGAACACCGATGGCGAAGGCAGGCTGCCAGCAGATTACTGACGCTGAGGTGCGAAGGTGCGGGGAGCGAACA"
## [26] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCGGACCTTTAAGTCAGCTGTGAAATACGGCGGCTCAACCGTCGAACTGCAGTTGATACTGGAGGTCTTGAGTGCACACAGGGATACTGGAATTCATGGTGTAGCGGTGAAATGCTCAGATATCATGAAGAACTCCAATCGCGAAGGCAGGTATCCGGGGTGCAACTGACGCTGAGGCTCGAAAGTGCGGGTATCAAAC"
## [27] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGGCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [28] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTGCGGGAGTGCGACTGACGCTGAAGCTCGAAGGTGCGGGTATCGAAC"
## [29] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGCACTGCAGCGCGAACTGGTCTTCTTGAGTACGCACAACGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACGGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [30] "GCGAGCGTTGTCCGGAATGACTGGGTGTAAAGGGAGCGTAGGCGGGATGGCAAGTCAGATGTGAAACCTGAGGGCTCAACCTTCAGACTGCATTTGAAACTGCTGTTCTTGAGTGAAGTAGAGGTAAGCGGAATTCCTGGTGTAGCGGTGAAATGCGTAGAGATCAGGAGGAACATCGGTGGCGAAGGCGGCTTACTGGGCTTTTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [31] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTTGCGGGAGTGCGACTGACGCTGAAGCTCGAAGGTGCGGGTATCGAAC"
## [32] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGTTAAGCGTGTCGTGAAATTCCGTCGCTCAACGGCGGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGACGGTAGCGCAACTGACGCTGAGGCTCGAAAGTGCGGGTATCGAAC"
## [33] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTTTGGTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGCCAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [34] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTGGTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGTCAGACTTGAGTGCGCGGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [35] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [36] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGTGTAGGTGGCCATGCAAGTCAGAAGTGAAAATCCGGGGCTCAACCCCGGAACTGCTTTTGAAACTGTAAGGCTGGAGTGCAGGAGGGGTGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTCACTGGACTGTAACTGACACTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [37] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGTTGCTCAACATCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [38] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCTGTCCTTTAAGCGTGTTGTGAAATGTAGGCGCTCAACGCCTGCACTGCAGCGCGAACTGAAGGACTTGAGTGTGCACGACGCTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGGCGAGAGCACAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [39] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAGAACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [40] "GCAAGCGTTATCCGGATTTACTGGGCGTAAAGGGAGCGTAGGCGGATATTTAAGTGGGATGTGAAATACCCGAGCTTAACTTGGGAGCTGCATTCCAAACTGGATATCTAGAGTGCAGGAGAGGAGAATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGAGATTAGGAAGAACACCAGTGGCGAAGGCGATTCTCTGGACTGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [41] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGCTAAGCGTGCCGTGAAATTCTGTCGCTCAACGGCAGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [42] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGCGCAGGCGGGCCGGCAAGTTGGAAGTGAAATCCGGGGGCTTAACCCCCGAACTGCTTTCAAAACTGCTGGTCTTGAGTGATGGAGAGGCAGGCGGAATTCCGTGTGTAGCGGTGAAATGCGTAGATATACGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACATTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [43] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [44] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGACGCTCAACGTCTGCACTGCAGCGCGAACTGGTTTCCTTGAGTACGCACAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTGACGGTAGCGCAACTGACGCTGAGGCTCGAAAGCGCGGGTATCGAAC"
## [45] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTTATAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGTGAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [46] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTGTTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGGCAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGTAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [47] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGGAGATTAAGCGTGTTGTGAAATGTAGATGCTCAACATCTGAACTGCAGCGCGAACTGGTTTCCTTGAGTACGCATAAAGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [48] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGCCCCTTAAGCGTGTTGTGAAATGCCGCGGCTCAACCGTGGCACTGCAGCGCGAACTGGGGGGCTTGAGTGCACGCAACGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [49] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGAGTAGGCGGATTGGCAAGTTGGGAGTGAAATGTCGGGGCTTAACCCCGGAACTGCTTCCAAAACTGTTGATCTTGAGTGATGGAGAGGCAGGCGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGGCCTGCTGGACATTAACTGACGCTGAGGAGCGAAAGCGTGGGGAGCAAAC"
## [50] "GCAAGCGTTAATCGGAATAACTGGGCGTAAAGCGCACGTAGGTGGTTCGACAAGTCAGATGTGAAAGCCCCGGGCTTAACCTGGGATGTGCATTTGAAACTGTTGGACTCGAGTACTGTAGAGGGAGGTAGAATTCCAGGTGTAGCGGTGAAATGCGTAGATATCTGGAGGAATACCAGTGGCGAAGGCGGCCTCCTGGACAGACACTGACACTGAGGTGCGAAAGCGTGGGGAGCAAAC"
## [51] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCTGTCCTTTAAGCGTGTTGTGAAATGTAGGCGCTCAACGCCTGCACTGCAGCGCGAACTGAAGGACTTGAGTGTGCACGACGCTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [52] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGGCCGTGGGTTAAGCGTGTCGTGAAATTCCGTCGCTCAACGGCGGACGTGCGGCGCGAACTGGTCCACTTGAGTACGCGGGACGTTGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTGGAGCGCAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [53] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGAGACAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGCCTTGCTAGAGTGCTGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [54] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGTGCGTAGGTCGCCGATTAAGTCAGCGGTGAAATCCAGTGGCTCAACCATCGGACTGCCGTTGAAACTGGCCGGCTTGAGTATGATTGAGGCAGGCGGAATGCGTGGTGTAGCGGTGAAATGCATAGATATCACGCAGAACCCCGATTGCGAAGGCAGCTTGCCAAGCCATGACTGACACTGAAGCACGAAAGCGTGGGTATCAAAC"
## [55] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCACAGCAAGTCTGATGTGAAAGCCCGGGGCCCAACCCCGGAACTGCATTGGAAACTGCTGGGCTTGAGTGCAGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [56] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCATGGCAAGTCTGATGTGAAAATCCCGGGCTCAACCCGGGAACTGCATTGGAAACTGTTAAGCTAGAGTGCAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACTGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [57] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTAAAGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCGCGGGACTGCTTTGGAAACTGTTTAACTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [58] "CCGGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTCTGTTAAGCGTGTTGTGAAATGTCGTGGCTCAACCGGGGCACTGCAGCGCGAACTGGCAGACTTGAGTGCACGGTAGGAAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCTTTCCGTAGTGTAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [59] "GCGAGCGTTGTCCGGAATGACTGGGCGTAAAGGGCGTGTAGGCGGCAGTATAAGTCCGGAGTGAAAGTCCTGCTTTCAAGGTGGGAATTGCTTTGGAGACTGTACAGCTTGAGTGCGGAAGAGGTAAGTGGAATTCCCAGTGTAGCGGTGAAATGCGTAGAGATTGGGAGGAACACCAGTGGCGAAGGCGACTTACTGGGCCGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCGAAC"
## [60] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGCATGCAAGTTGGATGTGAAATGTCACGGCTTAACCGTGGAGCTGCATCCAAAACTGCAAGTCTTGAGTGCCGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGCTTTCTGGACGGTAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [61] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGATGCAAGTCTGAAGTGAAATACCCGGGCTCAACCTGGGAACTGCTTTGGAAACTGTATTGCTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [62] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTTTTTTAAGCGTGTTGTGAAATACTGTCGCTCAACGACAGAGGTGCAGCGCGAACTGGAGGACTTGAGTGCGCGGAATGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCTTACAGTAGCGTAACTGACGCTGAAGCTCGAAAGTGCGGGTATCGAAC"
## [63] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGTGTAGGTGGCCAGGCAAGTCAGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTTGAAACTGCAGGGCTAGAGTGCAGGAGGGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACTGTAACTGACACTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [64] "GCAAGCGTTAATCGGAATCACTGGGCGTAAAGCGCACGTAGGCTGTTATGTAAGTCAGGGGTGAAATCCCACGGCTCAACCGTGGAACTGCCCTTGATACTGCACGACTTGAATCCGGGAGAGGGTGGCGGAATTCCAGGTGTAGGAGTGAAATCCGTAGATATCTGGAGGAACATCAGTGGCGAAGGCGGCCACCTGGACCGGTATTGACGCTGAGGTGCGAAAGCGTGGGGAGCAAAC"
## [65] "CCAGGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCCGTTTGGTAAGCGTGTTGTGAAATGTCGGGGCTCAACCTGGGCATTGCAGCGCGAACTGCCAGACTTGAGTGCGCAGGAAGTAGGCGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTGCTGTAGCGCAACTGACGCTGAAGCTCGAAAGCGTGGGTATCGAAC"
## [66] "GCAAGCGTTGTCCGGAATCATTGGGCGTAAAGAGTTCGTAGGCGGTTTGTTAAGTCTGGTGTTAAAGCCCGAAGCTCAACTTCGGTTCGGCACTGGATACTGGCAGACTAGAATGCGGTAGAGGTAAAGGGAATTCCTGGTGTAGCGGTGAAATGCGTAGATATCAGGAGGAACATCGGTGGCGTAAGCGCTTTACTGGGCCGTAATTGACGCTGAGGAACGAAAGCCAGGGTAGCGAAT"
## [67] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTTAAGCAAGTCAGAAGTGAAAGGCTGGGGCTCAACCCCGGGACTGCTTTTGAAACTGTTTAACTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [68] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCATGCAAGTTGGAAGTGAAAACTATGGGCTCAACCCATAGCCTGCTTTCAAAACTGCGTGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [69] "GCGAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGTGTAGGCGGGAAGGCAAGTCAGATGTGAAAACCAAAGGCTCAACCTTTGGCTTGCATTTGAAACTGTTTTTCTTGAGAGTGGGAGAGGTAAACGGAATTCCTAGTGTAGTAGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGTTTACTGGACCACAACTGACGCTGAGACACGAAAGCGTGGGGAGCAAAC"
## [70] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGCATGGTAAGTAAGATGTGAAAGCCCGAGGCTTAACCTCGAGGATTGCATTTTAAACTATCAAGCTAGAGTACAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGAAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCGAA"
## [71] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTCAAGCAAGTCAGAAGTGAAAGGCTGGGGCTCAACCCCGGGACTGCTTTTGAAACTGTTTGACTGGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [72] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGACGCAAGTCTGAAGTGAAATACCCGGGCTCAACCTGGGAACTGCTTTGGAAACTGTGTTGCTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAAGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [73] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGGAGAGCAAGTCAGCGGTGAAATACATGGGCTTAACCCATGGGCTGCCGTTGAAACTGTCCTTCTTGAGTGAAGTAGAGGCAAGCGGAATTCCGAGTGTAGCGGTGAAATGCGTAGATATTCGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGGCTTTTACTGACGCTGAGGCTCGAAAGTGTGGGGAGCAAAC"
## [74] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGTCTGGCAAGTCTGATGTGAAATCCCGGGGCTCAACCCTGGAACTGCATTGGAAACTGTCAGACTAGAGTGCCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [75] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTAAGCAAGTCAGAAGTGAAAGGCTGGGGCTCAACCCCGGGACTGCTTTTGAAACTGTTTGGCTAGAGTGCTGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACAGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [76] "GCAAGCGTTACTCGGAATTACTAGGCGTAAAGCGCGCGTAGGCGGAATGTTAAGTCTGTTGTGTAATCTCTGGGCTCAACCCAGAAACTGCAACAGAAACTGGCGTTCTTGAGTGAGGCAGAGGAAATCGGAATTCCTAGTGTAGCAGTGAAATGCGTAGATATTAGGAGGAACACCGGTGGCGAAGGCGGATTTCTGGGCCTTTACTGACGCTAAAGTGCGAAAGCTAGGGGAGCAAAC"
## [77] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGATGCCTAAGCGGGACCTCTAACCCGGGGGCTCAACCCCCGGCCGGGTCCCGAACTGGGCGTCTCGAGTGCGGTAGGGGCAGGTGGAATTCCATGTGTAGCGGTGGAATGCGCAGATATATGGAAGAACACCGACGGCGAAGGCAGCCTGCTGGGCCGACACTGACGCTGAGGTGCGAAAGCGCGGGGAGCGAACA"
## [78] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTAGTGCAAGTCTGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTATAACTAGAGTGTCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [79] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGTAGGCGGGAAGATAAGTTGGACGTCTAATCTATCGGCTCAACCGATAGTCGCGTTCAAAACTGTTTTTCTTGAGTGAAGTAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAAATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGGCTTTAACTGACGCTGAGGCTCGAAAGCGTGGGTAGCAAACA"
## [80] "GCGAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAAATTAAGTTAGTTGTGAAATCCCTCGGCTCAACTGAGGAACTGCAACTAAAACTGATTTTCTTGAGTACTGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACAGAAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [81] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTTGTAGGCGGTTTGTCGCGTCTGCTGTGAAAGGCCGGGGCTTAACCCCGTGTATTGCAGTGGGTACGGGCAGACTAGAGTGCAGTAGGGGAGACTGGAATTCCTGGTGTAGCGGTGGAATGCGCAGATATCAGGAGGAACACCGATGGCGAAGGCAGGTCTCTGGGCTGTAACTGACGCTGAGAAGCGAAAGCATGGGGAGCGAA"
## [82] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAATACAAGTTAGTTGTGAAATCCCTCGGCTTAACTGAGGAACTGCAACTAAAACTATATTTCTTGAGTGCTGGAGAGGAAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTTCTGGACAGTAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [83] "CCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGTGGATTGTTAAGTCAGTTGTGAAAGTTTGCGGCTCAACCGTAAAATTGCAGTTGAAACTGGCAGTCTTGAGTACAGTAGAGGTGGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACTCCGATTGCGAAGGCAGCTCACTAGACTGCAACTGACACTGATGCTCGAAAGTGTGGGTATCAAAC"
## [84] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGGCGGTATGGCAAGTCTGATGTGAAAGGCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTCACACTTGAGTGTCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGACAACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [85] "GCAAGCGTTATCCGGATTTATTGGGTGTAAAGGGTGCGTAGACGGGAAATTAAGTTAGTTGTGAAATCCCTCGGCTCAACTGAGGAACTGCAACTAAAACTGGTTTTCTTGAGTGCAGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGACTTACTGGACTGTAACTGACGTTGAGGCACGAAAGTGTGGGGAGCAAAC"
## [86] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAAACCATGGGCTCAACCCATGAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGTCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [87] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTGTGTAAGTCTGAAGTGAAAGCCCGGGGCTCAACCGCGGGACTGCTTTGGAAACTATGCAGCTAGAGTGTCGGAGAGGTAAGTGGAATTCCCAGTGTAGCGGTGAAATGCGTAGATATTGGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [88] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCGCTCGAGCGGGACCTCTAACCCGGGGGCTCAACCTCCGGCCGGGTCCCGGACCGTGCGGCTCGGGTGCGGTAGGGGCAGGCGGAACTCCAAGTGTAGCGGTGAAATGCGCAGATATTTGGAGGAACACCGATGGCGAAGGCAGCCTGCTGGGCCGCCACCGACGCTGAGGCGCGAAAGCCGGGGGAGCGAACA"
## [89] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGTGGTATGGCAAGTCAGAAGTGAAAACCCAGGGCTTAACTCTGGGACTGCTTTTGAAACTGTCAGACTAGAGTGCAGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCTTACTGGACTGAAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [90] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTGTGGCAAGTCTGATGTGAAAGGCATGGGCTCAACCTGTGGACTGCATTGGAAACTGTCATACTTGAGTGCCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [91] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCGACGGCGAAGGCAGCTCTCTGGGCCGAAACTGACGCTGAGGCGCGAAAGCTGGGGGAGCGAAC"
## [92] "CCAAGCGTTATCCGGATTTATTGGGCGTAAAGCGAGCGCAGACGGTTATTTAAGTCTGAAGTGAAAGCCCTCAGCTCAACTGAGGAATTGCTTTGGAAACTGGATGACTTGAGTGCAGTAGAGGAAAGTGGAACTCCATGTGTAGCGGTGAAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACGTTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [93] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGTACGTAGGTGGTTACCTAAGCACGAGGTATAAGGCAATGGCTTAACCATTGTTCGCCTTGTGAACTGGGCTACTTGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACACTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [94] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCACGGCAAGCCAGATGTGAAAGCCCGGGGCTCAACCCCGGGACTGCATTTGGAACTGCTGAGCTAGAGTGTCGGAGAGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [95] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGAGTACGTAGGTGGTTTTCTAAGCACGGGGTTTAAGGCAATGGCTTAACCATTGTTCGCCTTGTGAACTGGAAGACTTGAGTGCAGGAGAGGAAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTTCTGGACTGTAACTGACACTGAGGTACGAAAGCGTGGGGAGCAAACA"
## [96] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTTTGCAAGTCTGATGTGAAAGGCGGGGGCTCAACCCCTGGACTGCATTGGAAACTGTGAGGCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [97] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGACCGGCAAGTTGGAAGTGAAAACTATGGGCTCAACCCATAAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGACCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [98] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGACCGGCAAGTTGGAAGTGAAAACCATGGGCTCAACCCGTGAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGACCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [99] "GCGAGCGTTATCCGGAATCATTGGGCGTAAAGCGCGCGCAGGCGGGCTTTCAAGCGGCGGCGTCGAAGCCGGGGGCTCAACCCCCGGAAGCGCCCCGAACTGGAAGCCTCGGATGCGGCAGGGGGAGGCGGAATTCCCGGTGTAGCGGTGAAATGCGCAGATATCGGGAAGAACACCGACGGCGAAGGCAGCCTCCTGGGCCGGCATCGACGCTGAGGCGCGAAAGCTGGGGGAGCGAAC"
## [100] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGAGCGCTAAGCGGGACCTCTAACCCGAGGGCTCAACCCCCGGCCGGGTCCCGAACTGGCGCTCTCGAGTGCGGTAGGGGAGAGCGGAATTCCCGGTGTAGCGGTGGAATGCGCAGATATCGGGAAGAACACCGACGGCGAAGGCAGCTCTCTGGGCCGAAACTGACGCTGAGGCGCGAAAGCTGGGGGAGCGAACA"
## [101] "GCTAGCGTTATCCGGATTTACTGGGCGTAAAGGGTGCGTAGGCGGTCTTTTAAGTCAGGAGTGAAAGGCTACGGCTCAACCGTAGTAAGCTCTTGAAACTGGAGGACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTAGCGAAGGCGGCTCTCTGGACTGTAACTGACGCTGAGGCACGAAAGCGTGGGGAGCAAACA"
## [102] "GCAAGCGTTATCCGGAATTACTGGGTGTAAAGGGTGCGTAGGTGGTATGGCAAGTCAGAAGTGAAAACCCAGGGCTTAACTCTGGGACTGCTTTTGAAACTGTCAGACTGGAGTGCAGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACATCAGTGGCGAAGGCGGCTTACTGGACTGAAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAAC"
## [103] "GCGAGCGTTATCCGGAATGATTGGGCGTAAAGCGCGCGCAGGCGGCCGCTCAAGCGGGACCTCTAACCCCGGGGCTCAACCCCGGGCCGGGTCCCGAACTGGGCGGCTCGAGTGCGGTAGGGGAGAGCGGAATTCCAAGTGTAGCGGTGAAATGCGCAGATATTTGGAAGAACACCGATGGCGAAGGCAGCTCTCTGGGCCGTCACTGACGCTGAGGCGCGAAAGCCGGGGGAGCGAACA"
## [104] "GCGAGCGTTGTCCGGAATTATTGGGCGTAAAGGGCTTGTAGGCGGTTGGTCGCGTCTGCCGTGAAATCCTCTGGCTTAACTGGGGGCGTGCGGTGGGTACGGGCTGACTTGAGTGCGGTAGGGGAGACTGGAACTCCTGGTGTAGCGGTGGAATGCGCAGATATCAGGAAGAACACCGGTGGCGAAGGCGGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
## [105] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGAATGGCAAGTCTGATGTGAAAGGCCGGGGCTCAACCCCGGGACTGCATTGGAAACTGTCAATCTAGAGTACCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [106] "TCTAGTGGTAGCAGTTTTTATTGGGCCTAAAGCGTCCGTAGCCGGTTTAATAAGTCTCTGGTGAAATCCTGCAGCTTAACTGTGGGAATTGCTGGAGATACTATTAGACTTGAGATCGGGAGAGGTTAGAGGTACTCCCAGGGTAGAGGTGAAATTCTGTAATCCTGGGAGGACCGCCTGTTGCGAAGGCGTCTGACTGGAACGATTCTGACGGTGAGGGACGAAAGCTAGGGGCGCGAA"
## [107] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGTATGGCAAGTCTGATGTGAAAGGCCAGGGCTCAACCCTGGGACTGCATTGGAAACTGTCGAACTAGAGTGTCGGAGAGGCAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [108] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGAGCGCTAAGCGGGACCTCTAACCCGAGGGCTCAACCCCCGGCCGGGTCCCGAACTGGCGCTCTCGAGTGCGGTAGGGGAGAGCGGAATTCCCGGTGTAGCGGTGGAATGCGCAGATATCGGGAGGAACACCGACGGCGAAGGCAGCTCTCTGGGCCGAAACTGACGCTGAGGCGCGAAAGCTGGGGGAGCGAACA"
## [109] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCTGTGCAAGTCTGAAGTGAAAGGCATGGGCTCAACCTGTGGACTGCTTTGGAAACTGTGCAGCTAGAGTGTCGGAGAGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATGACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [110] "GCTAGCGTTATCCGGAATTACTGGGCGTAAAGGGTGCGTAGGTGGTTTCTTAAGTCAGAGGTGAAAGGCTACGGCTCAACCGTAGTAAGCCTTTGAAACTGGGAAACTTGAGTGCAGGAGAGGAGAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTTGCGAAGGCGGCTCTCTGGACTGTAACTGACACTGAGGCACGAAAGCGTGGGGAGCAAACA"
## [111] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGTCAAGCGGGGTTTCAAATCCAGGGGCTCAACCTCTGGCCGGACCCCGAACTGGCGGGCTCGAGTGCGGTAGAGGAAGGTGGAATTCCCAGTGTAGCGGTGAAATGCGCAGATATTGGGAAGAACACCGATGGCGAAGGCAGCCTTCTGGGCCGCCACTGACGCTGAGGCGCGAAAGCTAGGGGAGCGAACA"
## [112] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCGCAGCAAGTCTGATGTGAAAGGCAGGGGCTTAACCCCTGGACTGCATTGGAAACTGCTGTGCTTGAGTGCCGGAGGGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [113] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGATTAGCAAGTCTGATGTGAAAGGCAGGGGCTCAACCCCTGGACTGCATTGGAAACTGCCAGTCTTGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGCAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [114] "TCAAGCGTTGTTCGGAATCACTGGGCGTAAAGCGTGCGTAGGCTGTTTCGTAAGTCGTGTGTGAAAGGCGCGGGCTCAACCCGCGGACGGCACATGATACTGCGAGACTAGAGTAATGGAGGGGGAACCGGAATTCTCGGTGTAGCAGTGAAATGCGTAGATATCGAGAGGAACACTCGTGGCGAAGGCGGGTTCCTGGACATTAACTGACGCTGAGGCACGAAGGCCAGGGGAGCGAAA"
## [115] "GCGAGCGTTATCCGGATTCATTGGGCGTAAAGCGCGCGTAGGCGGCCCGGCAGGCCGGGGGTCGAAGCGGGGGGCTCAACCCCCCGAAGCCCCCGGAACCTCCGCGGCTTGGGTCCGGTAGGGGAGGGTGGAACACCCGGTGTAGCGGTGGAATGCGCAGATATCGGGTGGAACACCGGTGGCGAAGGCGGCCCTCTGGGCCGAGACCGACGCTGAGGCGCGAAAGCTGGGGGAGCGAAC"
## [116] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGACCGGCAAGTTGGAAGTGAAATCCATGGGCTCAACCCGTGAATTGCTTTCAAAACTGCTGGCCTTGAGTAGTGCAGAGGTAGGTGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGACCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [117] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAAACCATGGGCTCAACCCATGAATTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGATGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGTCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [118] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCCGCGCCGGGTACGGGCGGGCTTGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
## [119] "GCAAGCGTTATCCGGAATTATTGGGCGTAAAGGGCTCGTAGGCGGTTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCAATGGCGAAGGCAGGTCTCTGGGCCGTTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAAC"
##
## $xy
## [1] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCCGTGCAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGTATGGCTGGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [2] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGACGGCACTGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGTAGAGCTAGAGTGCTGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [3] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGCGTAGGCGGGGATGCAAGTCAGATGTGAAATCTATGGGCTTAACCCATAAACTGCATTTGAAACTGTATCTCTTGAGTGCTGGAGAGGTAGACGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGTCTACTGGACAGTAACTGACGCTGAGGCGCGAGAGCGTGGGGAGCAAAC"
##
## $xz
## character(0)
##
## $yz
## [1] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCGATCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGGTCGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [2] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [3] "GCGAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTTGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTCCAAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [4] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTCGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCGAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [5] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGAAGGACAAGTTGGAAGTGAAACCCACGGGCTCAACCCGTGAACTGCTTTCAAAACTGTTTTTCTTGAGTGGTGTAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [6] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGACGGGACTTTAAGTCAGCTGTGAAATTTTCCGGCTCAACCGGGAAACTGCAGTTGATACTGGCGTCCTTGAGTACGGTCGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACCCCGATTGCGAAGGCAGCCTGCCAGACCGCAACTGACGTTCATGCTCGAAAGTGCGGGTATCAAAC"
## [7] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCGGGCTGTTAAGTCAGCGGTCAAATGTCAGGGCCCAACCTTGGCATGCCGTTGATACTGGCGGCCTTGAGTTCACACAAGGAAGGTGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTTCTGGGGTGTTACTGACGCTGAGGCTCGAAAGTGCGGGAATCAAACA"
## [8] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGAAAGCAAGTCAGATGTGAAAACTGTGGGCTCAACCCACAGCCTGCATTTGAAACTGTTTTTCTTGAGTACTGGAGAGGCAGATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGATCTGCTGGACAGCAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [9] "GCAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGCAGGCGGTTCATCAAGTAGGATGTGAAATCCCCGGGCTCAACCTGGGAACAGCATACTAAACTGGTGGACTAGAGTATTGCAGGGGGAGACGGAATTCCAGGTGTAGCGGTGGAATGCGTAGATATCTGGAAGAACACCAAAGGCGAAGGCAGTCTCCTGGGCAAATACTGACGCTCATATGCGAAAGCGTGGGTAGCAAAC"
##
## $xy_only
## [1] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGTAGACGGCCGTGCAAGTCTGATGTGAAAGGCTGGGGCTCAACCCCGGGACTGCATTGGAAACTGTATGGCTGGAGTGCCGGAGAGGTAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [2] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGACGGCACTGCAAGTCTGAAGTGAAAGCCCGGGGCTCAACCCCGGGACTGCTTTGGAAACTGTAGAGCTAGAGTGCTGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGGTAACTGACGTTGAGGCTCGAAAGCGTGGGGAGCAAAC"
## [3] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGCGTAGGCGGGGATGCAAGTCAGATGTGAAATCTATGGGCTTAACCCATAAACTGCATTTGAAACTGTATCTCTTGAGTGCTGGAGAGGTAGACGGAATTCCTTGTGTAGCGGTGAAATGCGTAGATATAAGGAAGAACACCAGTGGCGAAGGCGGTCTACTGGACAGTAACTGACGCTGAGGCGCGAGAGCGTGGGGAGCAAAC"
##
## $xz_only
## character(0)
##
## $yz_only
## [1] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGCGATCAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGGTCGTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [2] "ACAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGGAAGACAAGTTGGAAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCAAAACTGTTTTTCTTGAGTAGTGCAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACCAACTGACGCTGAGGCTCGAAAGTGTGGGTAGCAAAC"
## [3] "GCGAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTTGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTCCAAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [4] "GCAAGCGTTGTCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGAGATGCAAGTCGGGAGTGAAATCCATGGGCTCAACCCATGAACTGCTTTCGAAACTGTATCCCTTGAGTATCGGAGAGGCAAGCGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTGCTGGACGACAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [5] "GCAAGCGTTGTCCGGAATTACTGGGTGTAAAGGGAGCGCAGGCGGAAGGACAAGTTGGAAGTGAAACCCACGGGCTCAACCCGTGAACTGCTTTCAAAACTGTTTTTCTTGAGTGGTGTAGAGGTAGGCGGAATTCCCGGTGTAGCGGTGGAATGCGTAGATATCGGGAGGAACACCAGTGGCGAAGGCGGCCTACTGGGCACTAACTGACGCTGAGGCTCGAAAGCATGGGTAGCAAAC"
## [6] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGCAGACGGGACTTTAAGTCAGCTGTGAAATTTTCCGGCTCAACCGGGAAACTGCAGTTGATACTGGCGTCCTTGAGTACGGTCGAGGCAGGCGGAATTCGTGGTGTAGCGGTGAAATGCTTAGATATCACGAAGAACCCCGATTGCGAAGGCAGCCTGCCAGACCGCAACTGACGTTCATGCTCGAAAGTGCGGGTATCAAAC"
## [7] "GCGAGCGTTATCCGGATTTATTGGGTTTAAAGGGAGCGTAGGCGGGCTGTTAAGTCAGCGGTCAAATGTCAGGGCCCAACCTTGGCATGCCGTTGATACTGGCGGCCTTGAGTTCACACAAGGAAGGTGGAATTCGTCGTGTAGCGGTGAAATGCTTAGATATGACGAAGAACTCCGATTGCGAAGGCAGCCTTCTGGGGTGTTACTGACGCTGAGGCTCGAAAGTGCGGGAATCAAACA"
## [8] "GCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGCGTGTAGGCGGGAAAGCAAGTCAGATGTGAAAACTGTGGGCTCAACCCACAGCCTGCATTTGAAACTGTTTTTCTTGAGTACTGGAGAGGCAGATGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGATCTGCTGGACAGCAACTGACGCTGAGGCGCGAAAGCGTGGGGAGCAAAC"
## [9] "GCAAGCGTTAATCGGAATAACTGGGCGTAAAGGGCATGCAGGCGGTTCATCAAGTAGGATGTGAAATCCCCGGGCTCAACCTGGGAACAGCATACTAAACTGGTGGACTAGAGTATTGCAGGGGGAGACGGAATTCCAGGTGTAGCGGTGGAATGCGTAGATATCTGGAAGAACACCAAAGGCGAAGGCAGTCTCCTGGGCAAATACTGACGCTCATATGCGAAAGCGTGGGTAGCAAAC"
##
## $xyz
## character(0)#--------------------------------------------------------------------------------------------------------------
#FIGURE 2B, C, D: ANCOM NEG-UNTREATED
#--------------------------------------------------------------------------------------------------------------
#Transform count data in the phyloseq object
ps_gg_fp_f_prevalence_filter_2019_05_26_proportion<-transform_sample_counts(ps_gg_fp_f_prevalence_filter_2019_05_26, function(x)(x/sum(x)))## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'###RUN ANCOM###
#US
dataset<-ps_gg_fp_f_prevalence_filter_2019_05_26
metadata<-as.data.frame(sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26))
metadata<-metadata[metadata$hiv_phenotype %in% c("1_hiv_negative", "4_unsuppressed"), , drop=F]
metadata<-as.data.frame(as.matrix(metadata[metadata$sexual_orientation != "MSM" | is.na(metadata$sexual_orientation), , drop=F]))
metadata_boston<-metadata[metadata$sample_cohort == "boston", , drop=F]
sample_data(dataset)<-metadata_boston## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'ps.taxa.sub <- phyloseq::prune_taxa(taxa_sums(dataset) > 0, dataset)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'out <- ANCOMBC::ancombc(data = ps.taxa.sub, formula = "hiv_phenotype", tax_level = NULL,
p_adj_method = "BH", prv_cut = 0.05, lib_cut = 1000,
group = "hiv_phenotype", struc_zero = TRUE, neg_lb = FALSE, tol = 1e-5,
max_iter = 100, conserve = FALSE, alpha = 0.05, global = FALSE, n_cl = 6)## 'ancombc' is deprecated
## Use 'ancombc2' instead## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## `tax_level` is not speficified
## No agglomeration will be performed
## Otherwise, please speficy `tax_level` by one of the following:
## Kingdom, Phylum, Class, Order, Family, Genus, Species## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Warning: The group variable has < 3 categories
## The multi-group comparisons (global/pairwise/dunnet/trend) will be deactivated## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'res <- out$res
res_rn <- purrr::imap(res, function(x, y) dplyr::rename(x, !!y := hiv_phenotype4_unsuppressed))
res_df <- purrr::reduce(res_rn, dplyr::left_join, by = "taxon")
res_df <- dplyr::select(res_df, !starts_with("(Int"))
res_df_taxa <- dplyr::left_join(res_df, tibble::rownames_to_column(as.data.frame(phyloseq::tax_table(ps.taxa.sub))), by = c("taxon" = "rowname"))
res_df_taxa[["index_num"]] <- 1:nrow(res_df_taxa)
res_df_taxa[["cohort"]] <- "boston"
res_df_taxa[["method"]] <- "ancom"
res_df_taxa <- tidyr::unite(res_df_taxa, col = "Genus_Species", Genus, Species, index_num, remove = FALSE)
alpha = 0.05
taxa_sig <- dplyr::filter(res_df_taxa, q_val < 0.05)
taxa_sig$Genus_Species <- forcats::fct_reorder(taxa_sig$Genus_Species, taxa_sig$lfc, min)
taxa_sig$taxon_short <- stringr::str_sub(taxa_sig$taxon, 1, 4)
ps.taxa.rel.sig <- phyloseq::prune_taxa(taxa_sig[["taxon"]], ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'# Only keep filtered samples
ps.taxa.rel.sig <- phyloseq::prune_samples(rownames(phyloseq::otu_table(ps.taxa.sub)), ps.taxa.rel.sig)
sigtab_dataset_us <- taxa_sig
write.csv(sigtab_dataset_us, "ANCOM_US_NEG_UNTREAT_Filtered.csv")
#BOTSWANA
dataset<-ps_gg_fp_f_prevalence_filter_2019_05_26
metadata<-as.data.frame(sample_data(ps_gg_fp_f_prevalence_filter_2019_05_26))
metadata<-metadata[metadata$hiv_phenotype %in% c("1_hiv_negative", "4_unsuppressed"), , drop=F]
metadata<-as.data.frame(as.matrix(metadata[metadata$sexual_orientation != "MSM" | is.na(metadata$sexual_orientation), , drop=F]))
metadata_botswana<-metadata[metadata$sample_cohort == "botswana", , drop=F]
sample_data(dataset)<-metadata_botswana## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'ps.taxa.sub <- phyloseq::prune_taxa(taxa_sums(dataset) > 0, dataset)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'out <- ANCOMBC::ancombc(data = ps.taxa.sub, formula = "hiv_phenotype", tax_level = NULL,
p_adj_method = "BH", prv_cut = 0.05, lib_cut = 1000,
group = "hiv_phenotype", struc_zero = TRUE, neg_lb = FALSE, tol = 1e-5,
max_iter = 100, conserve = FALSE, alpha = 0.05, global = FALSE, n_cl = 6)## 'ancombc' is deprecated
## Use 'ancombc2' instead## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## `tax_level` is not speficified
## No agglomeration will be performed
## Otherwise, please speficy `tax_level` by one of the following:
## Kingdom, Phylum, Class, Order, Family, Genus, Species## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'## Warning: The group variable has < 3 categories
## The multi-group comparisons (global/pairwise/dunnet/trend) will be deactivated## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'## Also defined by 'tidytree'res <- out$res
res_rn <- purrr::imap(res, function(x, y) dplyr::rename(x, !!y := hiv_phenotype4_unsuppressed))
res_df <- purrr::reduce(res_rn, dplyr::left_join, by = "taxon")
res_df <- dplyr::select(res_df, !starts_with("(Int"))
res_df_taxa <- dplyr::left_join(res_df, tibble::rownames_to_column(as.data.frame(phyloseq::tax_table(ps.taxa.sub))), by = c("taxon" = "rowname"))
res_df_taxa[["index_num"]] <- 1:nrow(res_df_taxa)
res_df_taxa[["cohort"]] <- "botswana"
res_df_taxa[["method"]] <- "ancom"
res_df_taxa <- tidyr::unite(res_df_taxa, col = "Genus_Species", Genus, Species, index_num, remove = FALSE)
alpha = 0.05
taxa_sig <- dplyr::filter(res_df_taxa, q_val < 0.05)
taxa_sig$Genus_Species <- forcats::fct_reorder(taxa_sig$Genus_Species, taxa_sig$lfc, min)
taxa_sig$taxon_short <- stringr::str_sub(taxa_sig$taxon, 1, 4)
ps.taxa.rel.sig <- phyloseq::prune_taxa(taxa_sig[["taxon"]], ps_gg_fp_f_prevalence_filter_2019_05_26_proportion)## Found more than one class "phylo" in cache; using the first, from namespace 'phyloseq'
## Also defined by 'tidytree'# Only keep filtered samples
ps.taxa.rel.sig <- phyloseq::prune_samples(rownames(phyloseq::otu_table(ps.taxa.sub)), ps.taxa.rel.sig)
sigtab_dataset_botswana <- taxa_sig
write.csv(sigtab_dataset_botswana, "ANCOM_BOTS_NEG_UNTREAT_Filtered.csv")
# Save plots of individual ANCOM fold-change
ggsave("Figure2BCD_NEG_UNTREAT_ANCOM_US_v4.pdf", ggplot(data = sigtab_dataset_us, aes(x = Genus_Species, y = lfc)) + theme_bw() + coord_flip() + geom_bar(stat="identity", aes(fill = Genus)) + ggtitle("NEG-UNTREAT US") + theme(legend.position = "bottom"), width = 10, height = 10, units = "in", dpi = 300)
ggsave("Figure2BCD_NEG_UNTREAT_ANCOM_BOTS_v4.pdf", ggplot(data = sigtab_dataset_botswana, aes(x = Genus_Species, y = lfc)) + theme_bw() + coord_flip() + geom_bar(stat="identity", aes(fill = Genus)) + ggtitle("NEG-UNTREAT BOTSWANA") + theme(legend.position = "bottom"), width = 10, height = 10, units = "in", dpi = 300)
#Merge results from both sample cohorts:
sigtab_dataset_us_filter_prev_005 <- dplyr::filter(sigtab_dataset_us, abs(lfc) >= 0.05)
sigtab_dataset<-rbind(sigtab_dataset_us_filter_prev_005, sigtab_dataset_botswana)
sigtab_dataset[["index_num_all"]] <- 1:nrow(sigtab_dataset)
sigtab_dataset <- tidyr::unite(sigtab_dataset, col = "Genus_Species_all", Genus, Species, index_num_all, sep = "_", remove = FALSE)
sigtab_dataset <- tidyr::unite(sigtab_dataset, col = "Genus_Species_all_for_color", Genus, Species, sep = " ", remove = FALSE)
# Remove extra brackets around taxonomic name for labels
sigtab_dataset$Genus_label <- stringr::str_replace(sigtab_dataset$Genus, "^\\[([^\\]]+)\\]", "\\1")
sigtab_dataset$Species_label <- stringr::str_replace(sigtab_dataset$Species, "^\\[([^\\]]+)\\]", "\\1")
sigtab_dataset %>% dplyr::mutate(Genus_species_label =
dplyr::case_when(is.na(stringr::str_extract(Genus_label, "\\[")) & is.na(stringr::str_extract(Species_label, "\\[")) ~ paste(Genus_label,Species_label),
TRUE ~ paste(Genus_label,"sp"))) -> sigtab_dataset
sigtab_dataset$Genus_species_label <- stringr::str_replace(sigtab_dataset$Genus_species_label, "\\[([^\\]]+)\\]", "")
sigtab_dataset$Genus_species_label <- stringr::str_replace(sigtab_dataset$Genus_species_label, "\\ +", " ")
sigtab_dataset <- dplyr::arrange(sigtab_dataset, cohort, lfc)
sigtab_dataset$Genus_Species_all<-factor(sigtab_dataset$Genus_Species_all, levels = c(sigtab_dataset$Genus_Species_all))
# Make lookup table for Genus_species_label
Genus_species_label_lookup <- dplyr::distinct(sigtab_dataset, Genus_Species_all, .keep_all = TRUE)$Genus_species_label
names(Genus_species_label_lookup) <- dplyr::distinct(sigtab_dataset, Genus_Species_all, .keep_all = TRUE)$Genus_Species_all
#Load color dictionary and construct figure:
dictionary_unique <- read.csv("COLOR_DICTIONARY3.csv", sep=";")
sigtab_dataset$Genus_species_label <- factor(sigtab_dataset$Genus_species_label, levels = unique(sigtab_dataset$Genus_species_label))
dictionary_plot <- dictionary_unique[dictionary_unique$Genus_species_label%in%sigtab_dataset$Genus_species_label,,drop=F]
rownames(dictionary_plot)<-dictionary_plot$Genus_species_label
dictionary_plot<-dictionary_plot[as.character(unique(sigtab_dataset$Genus_species_label)),,drop=F]
sigtab_dataset$cohort <- fct_relevel(as.factor(sigtab_dataset$cohort), "boston", "botswana")
header_namer <- as_labeller(c(`boston` = "U.S.\nHIV-uninfected HIV+ Untreated", `botswana` = "Botswana\nHIV-uninfected HIV+ Untreated"))
ggsave("Figure2BCD_NEG_UNTREAT_ANCOM_ALL_v7.pdf",
ggplot(data = sigtab_dataset, aes(x = Genus_Species_all, y = lfc)) +
geom_bar(stat = "identity", aes(fill = Genus_species_label)) +
scale_fill_manual(values = as.character(dictionary_plot$Color)) +
coord_flip() + scale_x_discrete(label = as_labeller(Genus_species_label_lookup)) + theme_bw() +
theme(legend.position = "bottom", axis.text.x = element_text(size = 16), axis.text.y = element_text(size = 6, face = "plain"), axis.ticks.y = element_blank(),
panel.border = element_rect(linetype = "solid", fill = NA, linewidth = 1), axis.title.y = element_blank(),
strip.background = element_rect(colour = "white", fill = "white"), strip.text = element_text(colour = "black", face = "bold", size = rel(1.2))) +
geom_hline(yintercept = 0) + ylab("log2FoldChange") +
facet_wrap(vars(cohort), scales = "fixed", ncol = 3, labeller = labeller(cohort = header_namer)), width = 15, height = 20, units = "in", dpi = 300)
#--------------------------------------------------------------------------------------------------------------