Mercurial > repos > petr-novak > dante_ltr

annotate clean_ltr.R @ 6:b91ca438a1cb draft

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"planemo upload commit 9633fb98932151f059ce02a0ce202a4374ef8d68"
author petr-novak
date Thu, 19 May 2022 08:21:55 +0000
parents 93d35ae65e1b
children c33d6583e548
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1 #!/usr/bin/env Rscript
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2 initial_options <- commandArgs(trailingOnly = FALSE)
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3 file_arg_name <- "--file="
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4 script_name <- normalizePath(sub(file_arg_name, "", initial_options[grep
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5 (file_arg_name,
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6 initial_options)]))
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7 script_dir <- dirname(script_name)
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8 library(optparse)
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9
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10 parser <- OptionParser()
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11 option_list <- list(
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12 make_option(c("-g", "--gff3"), action = "store", type = "character",
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13 help = "gff3 with LTR Transposable elements", default = NULL),
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14 make_option(c("-s", "--reference_sequence"), action = "store", type = "character",
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15 help = "reference sequence as fasta",
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16 default = NULL),
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17 make_option(c("-o", "--output"), action = "store", type = "character",
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18 help = "output file prefix", default = NULL),
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19 make_option(c("-c", "--cpu"), type =
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20 "integer", default = 5, help = "Number of cpu to use [default %default]",
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21 metavar = "number")
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22
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23 )
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24 description <- paste(strwrap(""))
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25
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26 epilogue <- ""
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27 parser <- OptionParser(option_list = option_list, epilogue = epilogue, description =
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28 description, usage = "usage: %prog COMMAND [OPTIONS]")
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29 opt <- parse_args(parser, args = commandArgs(TRUE))
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30
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31
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32 # load packages
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33 suppressPackageStartupMessages({ library(rtracklayer)
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34 library(Biostrings)
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35 library(BSgenome)
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36 library(parallel)
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37 })
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38
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39 # CONFIGURATION
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40 # load configuration files and functions:
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41 lineage_file <- paste0(script_dir, "/databases/lineage_domain_order.csv")
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42 ltr_utils_r <- paste0(script_dir, "/R/ltr_utils.R")
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43 if (file.exists(lineage_file)) {
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44 lineage_info <- read.table(lineage_file, sep = "\t",
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45 header = TRUE,
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46 as.is = TRUE)
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47 source(ltr_utils_r)
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48 }else {
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49 lineage_file <- paste0(script_dir, "/.
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50 ./share/dante_ltr/databases/lineage_domain_order.csv")
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51 ltr_utils_r <- paste0(script_dir, "/.
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52 ./share/dante_ltr/R/ltr_utils.R")
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53 if (file.exists(lineage_file)) {
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54 lineage_info <- read.table(lineage_file, sep = "\t",
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55 header = TRUE,
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56 as.is = TRUE)
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57 source(ltr_utils_r)
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58 }else {
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59 (stop("configuration files not found"))
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60 }
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61 }
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62
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63
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64 ncpus <- opt$cpu
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65
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66
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67 # load data
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68 cat("reading fasta...")
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69 s <- readDNAStringSet(opt$reference_sequence) # genome assembly
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70 cat("done\n")
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71 outfile <- opt$output
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72 # clean sequence names:
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73 names(s) <- gsub(" .+", "", names(s))
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74 cat("reading gff...")
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75 g <- rtracklayer::import(opt$gff3, format = 'gff3') # DANTE gff3
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76 cat("done\n")
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77 # testing
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78 if (FALSE) {
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79 g <- rtracklayer::import("./test_data/big_test_data/dante_ltr_unfiltered_t.cacao.gff3")
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80 s <- readDNAStringSet("./test_data/big_test_data/T_cacao_chromosomes.fasta")
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81
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82 g <- rtracklayer::import("./test_data/sample_ltr_annotation.gff3")
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83 s <- readDNAStringSet("./test_data/sample_genome.fasta")
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84
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85 g <- rtracklayer::import("./test_data/DANTE_LTR_Vfaba_chr5.gff3")
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86 s <- readDNAStringSet("./test_data/211010_Vfaba_chr5.fasta")
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87 names(s) <- gsub(" .+", "", names(s))
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88 ncpus <- 10
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89 lineage_info <- read.table("databases/lineage_domain_order.csv", sep = "\t", header =
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90 TRUE, as.is = TRUE)
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91 source("./R/ltr_utils.R")
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92 }
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93
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94 ## ID in g must be unique - this could be a problem if gff is concatenated from multiple files!
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95 ## id ID is renamed - rename parent to!
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96 ## add chromosom index to disctinguish same IDs
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97 suffix <- as.numeric(seqnames(g))
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98 g$ID <- ifelse(is.na(g$ID), NA, paste0(g$ID,"_", suffix))
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99 g$Parent <- ifelse(is.na(g$Parent), NA, paste0(g$Parent,"_", suffix))
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100
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101 # get te sequence based on rank
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102
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103 # evaluate best TE - DLTP grou
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104 s_te <- get_te_sequences(g, s) # split by 'element quality'
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105 # best quality - split by lineage
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106 word_size <- 15
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107 # best TE
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108 TE_DLTP_info <- analyze_TE(s_te$DLTP, word_size = word_size, ncpus = ncpus)
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109
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110 # TE rank 2:
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111 TE_DLT_plus_DLP_info <- analyze_TE(c(s_te$DLP, s_te$DLT), word_size = word_size, ncpus
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112 = ncpus)
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113 TE_DLT_plus_DLP_info_DLTP_verified <- compare_TE_datasets(c(s_te$DLT, s_te$DLP), ncpus
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114 = ncpus,TE_DLTP_info$seqs_representative, word_size = word_size
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115 )
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116
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117 TE_DLT_plus_DLP_info_multiplicity <- verify_based_on_multiplicity(TE_DLT_plus_DLP_info)
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118 # create additional library from rank 2 verified by multiplicity
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119 id_for_additional_library <- setdiff(
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120 TE_DLT_plus_DLP_info_multiplicity$id_ok_mp_verified,
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121 TE_DLT_plus_DLP_info_DLTP_verified$id_ok_verified)
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122
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123 if (length(id_for_additional_library) > 1) {
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124 seqs_for_additional_library <- c(s_te$DLP, s_te$DLT)[names(c(s_te$DLP, s_te$DLT)) %in%
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125 id_for_additional_library]
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126 seqs_additional_info <- analyze_TE(seqs_for_additional_library, word_size =
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127 word_size, ncpus = ncpus)
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128 seq_representative <- c(
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129 TE_DLTP_info$seqs_representative,
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130 seqs_additional_info$seqs_representative
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131 )
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132 }else {
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133 if (length(id_for_additional_library) == 1) {
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134 seq_representative <- c(
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135 TE_DLTP_info$seqs_representative,
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136 c(s_te$DLP, s_te$DLT)[names(c(s_te$DLP, s_te$DLT)) %in% id_for_additional_library]
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137 )
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138 }else {
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139 seq_representative <- TE_DLTP_info$seqs_representative
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140 }
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141 }
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142
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143 # TE rank 3 - verify agains good DLTP
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144 TE_DL_info_DLTP_verified <- compare_TE_datasets(
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145 s_te$DL,
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146 TE_DLTP_info$seqs_representative, min_coverage = 0.95,
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147 ncpus = ncpus, word_size = word_size
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148 )
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149
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150
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151 R <- seq_diversity(seq_representative)$richness
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152 SI <- seq_diversity(seq_representative)$shannon_index
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153
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154 # final RM library:
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155 seq_representative_no_ssr <- seq_representative[R > 20]
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156
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157 ID <- g$ID[g$type == "transposable_element"]
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158 names(ID) <- paste0(seqnames(g), "_",
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159 start(g), "_",
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160 end(g), "#",
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161 g$Final_Classification
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162 )[g$type %in% "transposable_element"]
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163
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164 # create clean gff3
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165 id_of_good_te <- unique(c(
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166 TE_DLTP_info$te_conflict_info$ok,
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167 TE_DLT_plus_DLP_info_DLTP_verified$id_ok_verified,
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168 TE_DLT_plus_DLP_info_multiplicity$id_ok_mp_verified,
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169 TE_DL_info_DLTP_verified$id_ok_verified)
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170 )
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171
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172 c1 <- g$ID %in% ID[id_of_good_te]
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173 c2 <- sapply(g$Parent, function(x)ifelse(length(x) == 0, "", x)) %in% ID[id_of_good_te]
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174
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175 gff_out <- g[c1 | c2]
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176
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177 gff_te <- gff_out[gff_out$type %in% "transposable_element"]
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178 gff_5ltr <- gff_out[gff_out$LTR %in% "5LTR"]
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179 gff_3ltr <- gff_out[gff_out$LTR %in% "3LTR"]
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180
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181 full_te <- getSeqNamed(s, gff_te)
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182 names(full_te) <- paste0(gff_te$ID,":",names(full_te))
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183 ltr5 <- getSeqNamed(s, gff_5ltr)
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184 names(ltr5) <- paste0(gff_5ltr$Parent,":",names(ltr5))
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185 ltr3 <- getSeqNamed(s, gff_3ltr)
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186 names(ltr3) <- paste0(gff_3ltr$Parent,":",names(ltr3))
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187 inc <- gff_te$Rank != "DL"
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188
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189 writeXStringSet(seq_representative, paste0(opt$output, "_RM_lib_non_redundant.fasta"))
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190 writeXStringSet(full_te, paste0(opt$output, "_RM_lib_full_TE.fasta"))
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191 writeXStringSet(ltr5, paste0(opt$output, "_RM_lib_5LTR.fasta"))
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192 writeXStringSet(ltr3, paste0(opt$output, "_RM_lib_3LTR.fasta"))
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193
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194 export(gff_out, paste0(opt$output, "_clean.gff3"), format = "gff3")
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195
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196 lv <- sort(unique(gff_te$Final_Classification))
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197 te_count <- table(factor(gff_te$Final_Classification, levels=lv))
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198
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199 pdf(paste0(opt$output, "_summary.pdf"), width = 13, height=8, pointsize = 10)
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200 par(mfrow=c(1,2), mar=c(5,7,2,0), las=1)
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201 boxplot(width(gff_te) ~ factor(gff_te$Final_Classification, levels=lv),
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202 horizontal = TRUE, xlab="length[bp]", ylab="",
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203 names = paste0(gsub("^.+[|]", "", lv), " (", te_count, ")"),
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204 main = 'Full TE', at = seq_along(lv)*4
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205 )
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206 boxplot(width(gff_te[inc]) ~ factor(gff_te$Final_Classification[inc], levels=lv),
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207 horizontal = TRUE, xlab="length[bp]", ylab="",
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208 names = rep("", length(lv)),
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209 main = 'Full TE', at = seq_along(lv)*4-1, add=TRUE, col=2
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210 )
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211 par(mar=c(5,0,2,7))
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212 boxplot(width(gff_5ltr) ~ factor(gff_5ltr$Final_Classification, levels=lv),
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213 horizontal = TRUE, xlab="length[bp]", ylab="",
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214 names = rep("", length(lv)),
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215 main = "5'LTR", at = seq_along(lv)*4
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216 )
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217 boxplot(width(gff_5ltr[inc]) ~ factor(gff_5ltr$Final_Classification[inc], levels=lv),
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218 horizontal = TRUE, xlab="length[bp]", ylab="",
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219 names = rep("", length(lv)),
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220 main = "5'LTR", at = seq_along(lv)*4-1, add=TRUE, col=2
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221 )
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222 legend('bottomright', col=c("grey","2"), legend=c("All TE", "TE with PBS/TSD"), pch=15)
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223 dev.off()