Clean session and load packages
#clean session
rm(list = ls())
# unload all non-based packages
out <- sapply(paste('package:', names(sessionInfo()$otherPkgs), sep = ""), function(x) try(detach(x, unload = FALSE, character.only = TRUE), silent = T))
## vector with package names
x <- c( "pbapply", "parallel", "ggplot2", "ape", "seqinr", "ips","gtools", "seqmagick", "ggmsa", "cowplot"
)
aa <- lapply(x, function(y) {
# check if installed, if not then install
if (!y %in% installed.packages()[,"Package"])
install.packages(y)
# load package
try(require(y, character.only = T), silent = T)
})Functions and parameters
#functions and parameters
knitr::opts_knit$set(root.dir = normalizePath(".."))
knitr::opts_chunk$set(dpi = 50, fig.width = 12)
# ggplot2 theme
theme_set(theme_classic(base_size = 20))
split_seqs <- function(x) {
# from start to end
y <- sapply(1:20, function(y)
if (y == 1)
ifelse(substr(x, start = y + 1, stop = y + 1) == substr(x, start = y, stop = y), 0 , 1) else
ifelse(substr(x, start = y - 1, stop = y - 1) == substr(x, start = y, stop = y), 0 , 1)
)
x2 <- strsplit(x, "")[[1]]
w1 <- which(y == 1)
# to add single elements at the end
w1 <- c(w1, 21)
if (w1[1] == 1 & w1[2] == 2) w1 <- w1[2:length(w1)]
sgmts <- lapply(1:length(w1), function(w){
if (w1[w] == w1[1]) paste(x2[1:(w1[w] - 1)], collapse = "") else
paste(x2[w1[w - 1]:(w1[w] - 1)], collapse = "")
})
return(sgmts)
}
write_lek_fas <- function(x){
lk <- unlist(strsplit(x[seq(1, length(x), by = 2)], split = "_"))[seq(1, length(x), by = 2)]
lk <- gsub(">", "", paste0(lk, "_"))
out <- sapply(unique(lk), USE.NAMES = FALSE, function(y){
wch.lk <- grep(pattern = y, x = x)
fl.nms <-tempfile(tmpdir = getwd(), fileext = paste0("song.seq_",y, "SongsSeq.fa"))
writeLines(x[min(wch.lk):(max(wch.lk)+ 1)], con = fl.nms)
return(basename(fl.nms))
})
return(out)
}
# get leks with more than 5 song types
leks <- c("BR1", "CCE", "CCL", "HC1", "HC2", "LOC", "SAT", "SJA", "SUR", "TR1", "TR2")Create fastas for mafft
# create fastas for each lek
###----Alignment of phenotypic sequence data: CSV to fasta format----------------------
dat <- read.csv("./data/raw/segments_by_song_type.csv", stringsAsFactors = FALSE)
yrs <- read.csv("./data/raw/year_range_by_song_type.csv", stringsAsFactors = FALSE)
#Are id's unique?
length(dat$song.type) == length(unique(dat$song.type)) #OK
# seq as a single column
dat$seqs <- sapply(1:nrow(dat), function(i) paste(dat[i,3:22], collapse = "")
)
# one for each songtype year
dat.yr <- yrs
dat.yr$song.type.year <- paste(dat.yr$song.type, dat.yr$year, sep = "-")
dat.yr <- dat.yr[dat.yr$lek %in% leks, ]
dat.yr <- merge(dat.yr, dat[, c("song.type", "seqs")], by = "song.type")
# only 1 year per song type and using before 2008
dat.all.old <- dat.yr[order(dat.yr$song.type, dat.yr$year), ]
# only 1 year per song type and using before 2008
dat.early.old <- dat.all.old[!duplicated(dat.all.old$song.type), ]
# only 1 year per song type and exlcuding before 2008
dat.all.new <- dat.all.old[dat.all.old$year > 2007, ]
# only 1 year per song type and using before 2008
dat.early.new <- dat.early.old[dat.early.old$year > 2008, ]
# rest of song type years (without earliest)
dat.rest.old <- dat.all.old[!dat.all.old$song.type.year %in% dat.early.old$song.type.year,]
dat.rest.new <- dat.all.new[!dat.all.new$song.type.year %in% dat.early.new$song.type.year,]
# by lek all song types
for(i in leks){
# all old
write.fasta(sequences = as.list(dat.rest.old$seqs[dat.rest.old$lek == i]), names = dat.rest.old$song.type.year[dat.rest.old$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_rest_old_", i, ".fa"), open = "w", nbchar = 60, as.string = FALSE)
# early old
write.fasta(sequences = as.list(dat.early.old$seqs[dat.early.old$lek == i]), names = dat.early.old$song.type.year[dat.early.old$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_early_old_", i, ".fa"), open = "w", nbchar = 60, as.string = FALSE)
# all new
write.fasta(sequences = as.list(dat.rest.new$seqs[dat.rest.new$lek == i]), names = dat.rest.new$song.type.year[dat.rest.new$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_rest_new_", i, ".fa"), open = "w", nbchar = 60, as.string = FALSE)
# early new
write.fasta(sequences = as.list(dat.early.new$seqs[dat.early.new$lek == i]), names = dat.early.new$song.type.year[dat.early.new$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_early_new_", i, ".fa"), open = "w", nbchar = 60, as.string = FALSE)
}Aligning with mafft
# alignments real songs
# add using or not song types before 2008
use.old.fossils <- c("old", "new")
# type of costs
type <- c("optimal", "all.equal")
grd.costs <- expand.grid(use.old.fossils = use.old.fossils, type = type)
unq.sgmts <- c("d", "f", "m", "p", "s", "u")
asc.val <- c("0x73", "0x75", "0x70", "0x6D", "0x66", "0x64")
cost <- matrix(c(0.5, 1.25, 2, 0.5, 1.25, 2, # optimal
rep(c(1, 1, 1.5), 2)), # equal costs
nrow = 4, byrow = TRUE)
cost.mat <- data.frame(cost, stringsAsFactors = FALSE)
names(cost.mat)[1:3] <- c("different.category", "same.category", "same.segment")
cost.mat <- data.frame(cost.mat, grd.costs, stringsAsFactors = FALSE)
# grid cost values
grd.sgm <- expand.grid(sgmt1 = unq.sgmts, sgmt2 = unq.sgmts)
grd.sgm$type <- "different.category"
trlls <- c("f", "m", "s")
tones <- c("d", "p", "u")
grd.sgm$type[grd.sgm$sgmt1 %in% trlls & grd.sgm$sgmt2 %in% trlls] <- "same.category"
grd.sgm$type[grd.sgm$sgmt1 %in% tones & grd.sgm$sgmt2 %in% tones] <- "same.category"
grd.sgm$type[grd.sgm$sgmt1 == grd.sgm$sgmt2] <- "same.segment"
# create combs
cost.grds <- pblapply(1:nrow(cost.mat), cl = 3, function(x){
grd.sgm$cost <- NA
grd.sgm$cost[grd.sgm$type == "different.category"] <- cost.mat$different.category[x]
grd.sgm$cost[grd.sgm$type == "same.category"] <- cost.mat$same.category[x]
grd.sgm$cost[grd.sgm$type == "same.segment"] <- cost.mat$same.segment[x]
grd.sgm$pair <- paste(grd.sgm$sgmt1, "x", grd.sgm$sgmt2)
grd.sgm$asc1 <- grd.sgm$sgmt1
grd.sgm$asc2 <- grd.sgm$sgmt2
for(i in 1:length(unq.sgmts)){
grd.sgm$asc1 <- gsub(unq.sgmts[i], asc.val[i], grd.sgm$asc1)
grd.sgm$asc2 <- gsub(unq.sgmts[i], asc.val[i], grd.sgm$asc2)
}
grd.sgm$cost.type <- cost.mat$type[x]
grd.sgm$cost.vals <- cost.mat$cost.vals[x]
grd.sgm$use.old.fossils <- cost.mat$use.old.fossils[x]
return(grd.sgm)
})
length(cost.grds)
# get leks with more than 5 song types
# leks <- names(table(sgmnts$lek))[table(sgmnts$lek)> 5]
pboptions(type = "timer")
yrs <- read.csv("./data/raw/year_range_by_song_type.csv", stringsAsFactors = FALSE)
# exclude all equal
# cost.grds <- cost.grds[1]
out <- pblapply(1:length(cost.grds), cl = parallel::detectCores() - 1, function(y){
Y <- cost.grds[[y]]
w <- apply(Y[, c("asc1", "asc2", "cost")], 1, paste, collapse = " ")
writeLines(w, con = paste0("./data/processed/mafft_alignments/",Y$cost.type[1],"_cost_matrix.txt"))
out <- lapply(leks, function(i){
# don't do those with no data before 2008
if (min(yrs$year[yrs$lek == i]) > 2000 & Y$use.old.fossils[1] == "old")
return(NULL) else
{
cll <- paste0("mafft --localpair --maxiterate 50000 --textmatrix ./data/processed/mafft_alignments/",Y$cost.type[1] ,"_cost_matrix.txt ./data/processed/song_sequences/song_seqs_early_", Y$use.old.fossils[1], "_", i,".fa > ./data/processed/mafft_alignments/", i, "_", Y$cost.type[1], "_early_", Y$use.old.fossils[1], "_alignment.fa 2> ./data/processed/mafft_alignments/early_", Y$use.old.fossils[1], "_", i,"_", Y$cost.type[1], "_mafft_output.txt")
system(cll)
alg <- readLines(paste0("./data/processed/mafft_alignments/",i,"_", Y$cost.type[1], "_early_", Y$use.old.fossils[1], "_alignment.fa"))[2]
gaps <- nchar(alg) - 20
convg <- ifelse(any(grepl("Converged", readLines(paste0("./data/processed/mafft_alignments/early_", Y$use.old.fossils[1], "_", i ,"_", Y$cost.type[1],"_mafft_output.txt")))), "Y", "N")
df2 <- data.frame(lek = i, gaps, convg, cost.type = Y$cost.type[1], use.old.fossils = Y$use.old.fossils[1])
return(df2)
}
})
out <- out[!sapply(out, is.null)]
res <- do.call(rbind, out)
return(res)
})
res <- do.call(rbind, out)
write.csv(res, "./output/aligment_gaps_and_convergence_results.csv")Add new sequences to mafft alignments
#add new sequences only for optimal so far
alg <- list.files(path = "./data/processed/mafft_alignments", pattern = "alignment.fa$", full.names = TRUE)
# and new
rest.seqs <- list.files(path = "./data/processed/song_sequences", pattern = "rest", full.names = TRUE)
out <- lapply(leks, function(x){
# get alignments for x lek
lk.alg <- grep(x, alg, value = TRUE)
for(w in lk.alg){
cost_mat <- ifelse(grepl("optimal", w), "optimal", "all.equal")
use_old_fossils <- ifelse(grepl("old", w), "old", "new")
cll <- paste0("mafft --anysymbol --keeplength --add ", grep(paste0("new_", x), rest.seqs, value = TRUE)," --reorder ", file.path("./data/processed/mafft_alignments", paste(x, cost_mat, "early", use_old_fossils ,"alignment.fa", sep = "_")), " > ", file.path("./data/processed/mafft_alignments", paste(x, cost_mat, "all", use_old_fossils ,"alignment.fa", sep = "_")))
system(cll)
}
})Create fastas for prank
# create fastas for each lek
###----Alignment of phenotypic sequence data: CSV to fasta format----------------------
dat <- read.csv("./data/raw/segments_by_song_type.csv", stringsAsFactors = FALSE)
yrs <- read.csv("./data/raw/year_range_by_song_type.csv", stringsAsFactors = FALSE)
#Are id's unique?
length(dat$song.type) == length(unique(dat$song.type)) #OK
# seq as a single column
dat$seqs <- sapply(1:nrow(dat), function(i) paste(dat[i,3:22], collapse = "")
)
# replace song segments with bases
dat$seqs <- gsub(pattern = "f", replacement = "C", x = dat$seqs) # trill to pyrimidine
dat$seqs <- gsub(pattern = "s", replacement = "T", x = dat$seqs) # trill to pyrimidine
dat$seqs <- gsub(pattern = "m", replacement = "Y", x = dat$seqs) # medium trill treated as ambiguous between fast and slow
dat$seqs <- gsub(pattern = "u", replacement = "A", x = dat$seqs) # tone to purine
dat$seqs <- gsub(pattern = "d", replacement = "G", x = dat$seqs) # tone to purine
dat$seqs <- gsub(pattern = "p", replacement = "R", x = dat$seqs)
# one for each songtype year
dat.yr <- yrs
dat.yr$song.type.year <- paste(dat.yr$song.type, dat.yr$year, sep = "-")
dat.yr <- dat.yr[dat.yr$lek %in% leks, ]
dat.yr <- merge(dat.yr, dat[, c("song.type", "seqs")], by = "song.type")
# only 1 year per song type and using before 2008
dat.all.old <- dat.yr[order(dat.yr$song.type, dat.yr$year), ]
# only 1 year per song type and using before 2008
dat.early.old <- dat.all.old[!duplicated(dat.all.old$song.type), ]
# only 1 year per song type and exlcuding before 2008
dat.all.new <- dat.all.old[dat.all.old$year > 2000, ]
# only 1 year per song type and using before 2008
dat.early.new <- dat.early.old[dat.early.old$year > 2000, ]
# rest of song type years (without earliest)
dat.rest.old <- dat.all.old[!dat.all.old$song.type.year %in% dat.early.old$song.type.year,]
dat.rest.new <- dat.all.new[!dat.all.new$song.type.year %in% dat.early.new$song.type.year,]
# by lek all song types
for(i in leks){
# all old
write.fasta(sequences = as.list(dat.rest.old$seqs[dat.rest.old$lek == i]), names = dat.rest.old$song.type.year[dat.rest.old$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_rest_old_", i, "_prank.fa"), open = "w", nbchar = 60, as.string = FALSE)
# early old
write.fasta(sequences = as.list(dat.early.old$seqs[dat.early.old$lek == i]), names = dat.early.old$song.type.year[dat.early.old$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_early_old_", i, "_prank.fa"), open = "w", nbchar = 60, as.string = FALSE)
# all new
write.fasta(sequences = as.list(dat.rest.new$seqs[dat.rest.new$lek == i]), names = dat.rest.new$song.type.year[dat.rest.new$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_rest_new_", i, "_prank.fa"), open = "w", nbchar = 60, as.string = FALSE)
# early new
write.fasta(sequences = as.list(dat.early.new$seqs[dat.early.new$lek == i]), names = dat.early.new$song.type.year[dat.early.new$lek == i], file.out = paste0("./data/processed/song_sequences/song_seqs_early_new_", i, "_prank.fa"), open = "w", nbchar = 60, as.string = FALSE)
}Aligning with prank
prank_seqs <- list.files(path = "./data/processed/song_sequences", pattern = "prank")
# get only early ones
prank_seqs_early <- grep("early", prank_seqs, value = TRUE)
# get rest
prank_seqs_rest <- grep("rest", prank_seqs, value = TRUE)
out <- pblapply(prank_seqs_early, function(x) {
lek <- sapply(strsplit(x, "_", fixed = TRUE), "[[", 5)
all.fossils <- sapply(strsplit(x, "_", fixed = TRUE), "[[", 4)
old.fossils <- sapply(strsplit(x, "_", fixed = TRUE), "[[", 3)
cll <- paste0("prank -d=./data/processed/song_sequences/", x, " -o=./data/processed/mafft_alignments/", lek, "_prank_", old.fossils, "_", all.fossils,"_alignment.fa -iterate=100 -kappa=2")
system(cll)
})
# remove best.fas suffix
prank_algn <- list.files(path = "./data/processed/mafft_alignments", pattern = "best.fas", full.name = TRUE)
file.rename(from = prank_algn, to = gsub(".best.fas$", "", prank_algn))Add new sequences to prank alignments
#add new sequences only for optimal so far
alg <- list.files(path = "./data/processed/mafft_alignments", pattern = "prank", full.names = TRUE)
# and new
rest.seqs <- list.files(path = "./data/processed/song_sequences", pattern = "rest", full.names = TRUE)
rest.seqs <- grep("prank", rest.seqs, value = TRUE)
out <- lapply(leks, function(x){
# get alignments for x lek
lk.alg <- grep(x, alg, value = TRUE)
for(w in lk.alg){
cost_mat <- "prank"
use_old_fossils <- ifelse(grepl("old", w), "old", "new")
cll <- paste0("mafft --anysymbol --keeplength --add ", grep(paste0("new_", x), rest.seqs, value = TRUE)," --reorder ", file.path("./data/processed/mafft_alignments", paste(x, cost_mat, "early", use_old_fossils ,"alignment.fa", sep = "_")), " > ", file.path("./data/processed/mafft_alignments", paste(x, cost_mat, "all", use_old_fossils ,"alignment.fa", sep = "_")))
system(cll)
}
})Plot alignments
Mafft optimal costs:
- diff type = 0.5 (i.e. trill vs pure tone)
- same category = 1.25 (i.e. slow trill vs fast trill)
- same segment = 2 (i.e. slow trill vs slow trill)
Mafft agonostics (all equal) costs:
- diff type = 1.1 (i.e. trill vs pure tone)
- same category = 1.1 (i.e. slow trill vs fast trill)
- same segment = 1 (i.e. slow trill vs slow trill)
Prank: probabilistic multiple alignment program based on a novel algorithm that treats insertions correctly and avoids over-estimation of the number of deletion events
Make plots
#plot alignments
seqs <- list.files(path = "./data/processed/mafft_alignments", pattern = "\\.fa$", full.names = TRUE)
# remove those wtih repeated species
seqs <- grep("early", seqs, value = TRUE)
algn_plots <- lapply(seqs, function(i){
ggmsa(i, color = "Chemistry_AA") + ggtitle(gsub("_|\\.fa", " ", basename(i)))
})
names(algn_plots) <- basename(seqs)
algn_plots## $BR1_all.equal_early_new_alignment.fa
##
## $BR1_optimal_early_new_alignment.fa
##
## $BR1_prank_early_new_alignment.fa
##
## $BR1_prank_early_old_alignment.fa
##
## $CCE_all.equal_early_new_alignment.fa
##
## $CCE_all.equal_early_old_alignment.fa
##
## $CCE_optimal_early_new_alignment.fa
##
## $CCE_optimal_early_old_alignment.fa
##
## $CCE_prank_early_new_alignment.fa
##
## $CCE_prank_early_old_alignment.fa
##
## $CCL_all.equal_early_new_alignment.fa
##
## $CCL_all.equal_early_old_alignment.fa
##
## $CCL_optimal_early_new_alignment.fa
##
## $CCL_optimal_early_old_alignment.fa
##
## $CCL_prank_early_new_alignment.fa
##
## $CCL_prank_early_old_alignment.fa
##
## $HC1_all.equal_early_new_alignment.fa
##
## $HC1_all.equal_early_old_alignment.fa
##
## $HC1_optimal_early_new_alignment.fa
##
## $HC1_optimal_early_old_alignment.fa
##
## $HC1_prank_early_new_alignment.fa
##
## $HC1_prank_early_old_alignment.fa
##
## $HC2_all.equal_early_new_alignment.fa
##
## $HC2_all.equal_early_old_alignment.fa
##
## $HC2_optimal_early_new_alignment.fa
##
## $HC2_optimal_early_old_alignment.fa
##
## $HC2_prank_early_new_alignment.fa
##
## $HC2_prank_early_old_alignment.fa
##
## $LOC_all.equal_early_new_alignment.fa
##
## $LOC_optimal_early_new_alignment.fa
##
## $LOC_prank_early_new_alignment.fa
##
## $LOC_prank_early_old_alignment.fa
##
## $SAT_all.equal_early_new_alignment.fa
##
## $SAT_all.equal_early_old_alignment.fa
##
## $SAT_optimal_early_new_alignment.fa
##
## $SAT_optimal_early_old_alignment.fa
##
## $SAT_prank_early_new_alignment.fa
##
## $SAT_prank_early_old_alignment.fa
##
## $SJA_all.equal_early_new_alignment.fa
##
## $SJA_all.equal_early_old_alignment.fa
##
## $SJA_optimal_early_new_alignment.fa
##
## $SJA_optimal_early_old_alignment.fa
##
## $SJA_prank_early_new_alignment.fa
##
## $SJA_prank_early_old_alignment.fa
##
## $SUR_all.equal_early_new_alignment.fa
##
## $SUR_all.equal_early_old_alignment.fa
##
## $SUR_optimal_early_new_alignment.fa
##
## $SUR_optimal_early_old_alignment.fa
##
## $SUR_prank_early_new_alignment.fa
##
## $SUR_prank_early_old_alignment.fa
##
## $TR1_all.equal_early_new_alignment.fa
##
## $TR1_optimal_early_new_alignment.fa
##
## $TR1_prank_early_new_alignment.fa
##
## $TR1_prank_early_old_alignment.fa
##
## $TR2_all.equal_early_new_alignment.fa
##
## $TR2_optimal_early_new_alignment.fa
##
## $TR2_prank_early_new_alignment.fa
##
## $TR2_prank_early_old_alignment.fa
Save nexus files
### Convert fasta alignment to nexus
# read all alignments
all.algs <- list.files(pattern = ".fa$", path = "./data/processed/mafft_alignments/")
# loop over alignments
for (j in all.algs){
lek <- substring(j, 0, 3)
cost_mat <- if(grepl("optimal", j)) "optimal"
cost_mat <- if(grepl("all.equal", j)) "all.equal"
cost_mat <- if(grepl("prank", j)) "prank"
use_old_fossils <- ifelse(grepl("old", j), "old", "new")
# paste together call
cll <- paste0(" perl ./source/convertfasta2nex.pl ./data/processed/mafft_alignments/", j, " > ", "./data/processed/nexus/", gsub("\\.fa$", ".nex", j))
# call perl
system(cll)
}
# fix nexus header
Nex.file <-list.files(pattern = ".nex$", path = "./data/processed/nexus/", full.names = TRUE)
for(i in Nex.file){
align <- readLines(i)
align[4] <- 'format datatype=standard gap=- missing=? symbols="dfmpsu";'
# change seqs back to sound segments
if (grepl("prank", i)){
seqs <- align[7:(length(align)-2)]
sep <- separate(data.frame(seqs = seqs),col = "seqs",sep = "\t", into = c("song", "seqs"))
# replace song segments with bases
sep$seqs <- gsub(pattern = "C", replacement = "f", x = sep$seqs) # trill to pyrimidine
sep$seqs <- gsub(pattern = "T", replacement = "s", x = sep$seqs) # trill to pyrimidine
sep$seqs <- gsub(pattern = "Y", replacement = "m", x = sep$seqs) # medium trill treated as ambiguous between fast and slow
sep$seqs <- gsub(pattern = "A", replacement = "u", x = sep$seqs) # tone to purine
sep$seqs <- gsub(pattern = "G", replacement = "d", x = sep$seqs) # tone to purine
sep$seqs <- gsub(pattern = "R", replacement = "p", x = sep$seqs)
align[7:(length(align)-2)] <- paste0(sep$song, "\t", sep$seqs)
}
write(align, i)
}R session information
## R version 3.6.1 (2019-07-05)
## Platform: x86_64-pc-linux-gnu (64-bit)
## Running under: Ubuntu 18.04.4 LTS
##
## Matrix products: default
## BLAS: /usr/lib/x86_64-linux-gnu/atlas/libblas.so.3.10.3
## LAPACK: /usr/lib/x86_64-linux-gnu/atlas/liblapack.so.3.10.3
##
## locale:
## [1] LC_CTYPE=es_ES.UTF-8 LC_NUMERIC=C
## [3] LC_TIME=es_CR.UTF-8 LC_COLLATE=es_ES.UTF-8
## [5] LC_MONETARY=es_CR.UTF-8 LC_MESSAGES=es_ES.UTF-8
## [7] LC_PAPER=es_CR.UTF-8 LC_NAME=C
## [9] LC_ADDRESS=C LC_TELEPHONE=C
## [11] LC_MEASUREMENT=es_CR.UTF-8 LC_IDENTIFICATION=C
##
## attached base packages:
## [1] parallel stats graphics grDevices utils datasets methods
## [8] base
##
## other attached packages:
## [1] cowplot_1.0.0 ggmsa_0.0.2 seqmagick_0.1.3 gtools_3.8.1
## [5] ips_0.0.11 seqinr_3.6-1 ape_5.3 ggplot2_3.3.0
## [9] pbapply_1.4-2
##
## loaded via a namespace (and not attached):
## [1] Rcpp_1.0.3 lattice_0.20-41 tidyr_1.0.0
## [4] Biostrings_2.54.0 ggseqlogo_0.1 assertthat_0.2.1
## [7] zeallot_0.1.0 digest_0.6.22 R6_2.4.1
## [10] plyr_1.8.4 backports_1.1.5 stats4_3.6.1
## [13] evaluate_0.14 pillar_1.4.2 zlibbioc_1.32.0
## [16] rlang_0.4.1 lazyeval_0.2.2 phangorn_2.5.5
## [19] S4Vectors_0.24.3 Matrix_1.2-18 rmarkdown_1.17
## [22] labeling_0.3 stringr_1.4.0 igraph_1.2.4.1
## [25] munsell_0.5.0 compiler_3.6.1 xfun_0.12
## [28] pkgconfig_2.0.3 BiocGenerics_0.32.0 htmltools_0.4.0
## [31] tidyselect_0.2.5 tibble_2.1.3 quadprog_1.5-8
## [34] IRanges_2.20.2 XML_3.98-1.20 crayon_1.3.4
## [37] dplyr_0.8.3 withr_2.1.2 MASS_7.3-51.4
## [40] grid_3.6.1 nlme_3.1-142 jsonlite_1.6
## [43] gtable_0.3.0 lifecycle_0.1.0 magrittr_1.5
## [46] scales_1.0.0 tidytree_0.3.2 stringi_1.4.6
## [49] XVector_0.26.0 ellipsis_0.3.0 vctrs_0.2.0
## [52] fastmatch_1.1-0 tools_3.6.1 treeio_1.10.0
## [55] ade4_1.7-13 glue_1.3.1 purrr_0.3.3
## [58] yaml_2.2.1 colorspace_1.4-1 knitr_1.28