/
unlimited_breadsticks_0.1.sh
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unlimited_breadsticks_0.1.sh
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#!/bin/bash
# Unlimited Breadsticks Logo
echo "$(tput setaf 2)00000000000000000000000000"
echo "00000000000000000000000000"
echo "000000000$(tput setaf 4)^^^^^^^^$(tput setaf 2)000000000"
echo "000000$(tput setaf 4)^^^^^^^^^^^^^^$(tput setaf 2)000000"
echo "00000$(tput setaf 4)^^^$(tput setaf 3)UNLIMITED$(tput setaf 4)^^^^$(tput setaf 2)00000"
echo "00000$(tput setaf 4)^^$(tput setaf 3)BREADSTICKS$(tput setaf 4)^^^$(tput setaf 2)00000"
echo "00000$(tput setaf 4)^^^^^^^^^^^^^^^^$(tput setaf 2)00000"
echo "000000$(tput setaf 4)^^^^^^^^^^^^^^$(tput setaf 2)000000"
echo "000000000$(tput setaf 4)^^^^^^^^$(tput setaf 2)000000000"
echo "00000000000000000000000000"
echo "00000000000000000000000000$(tput sgr 0)"
echo " "
echo "version 0.1.1"
sleep 2s
# Setting input parameters
original_contigs=$1
run_title=$2
circ_length_cutoff=$3
linear_length_cutoff=$4
virus_domain_db=$5
LIN_MINIMUM_DOMAINS=$6
PROPHAGE=$7
FOR_PLASMIDS=$8
CENOTE_SCRIPT_DIR=$9
CIRC_MINIMUM_DOMAINS=${10}
MEM=${11}
CPU=${12}
base_directory=$PWD
echo "@@@@@@@@@@@@@@@@@@@@@@@@@"
echo "Your specified arguments:"
echo "original contigs: $original_contigs"
echo "title of this run: $run_title"
echo "minimum circular contig length: $circ_length_cutoff"
echo "minimum linear contig length: $linear_length_cutoff"
echo "virus domain database: $virus_domain_db"
echo "min. viral hallmarks for linear: $LIN_MINIMUM_DOMAINS"
echo "min. viral hallmarks for circular: $CIRC_MINIMUM_DOMAINS"
echo "Do Prophage Pruning?: $PROPHAGE"
echo "Filter out plasmids?: $FOR_PLASMIDS"
echo "Location of Cenote scripts: $CENOTE_SCRIPT_DIR"
echo "GB of memory: $MEM"
echo "number of CPUs available for run: $CPU"
echo "@@@@@@@@@@@@@@@@@@@@@@@@@"
if [ "$PROPHAGE" == "True" ] && [ $LIN_MINIMUM_DOMAINS -le 0 ] ; then
echo "Prophage pruning requires --lin_minimum_hallmark_genes >= 1. changing to:"
echo "--lin_minimum_hallmark_genes 1"
LIN_MINIMUM_DOMAINS=1
fi
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: locating inputs: " $MDYT
# looking for template file and contigs in working directory, or else copying them there
if [ -s ${base_directory}/${original_contigs} ] ; then
echo ${base_directory}/${original_contigs} ;
else
cp ${original_contigs} ${base_directory}/ ;
original_contigs=$( basename $original_contigs )
fi
# Making output folder
if [ ! -d "$run_title" ]; then
mkdir "$run_title"
else
rand_dir=$( head /dev/urandom | tr -dc A-Za-z0-9 | head -c 3 ; echo '' )
DAY1=$( date +"%m-%d-%y" )
mv ${run_title}/ ${run_title}_old_${DAY1}_${rand_dir}
mkdir "$run_title"
fi
if [ ${original_contigs: -3} == ".fa" ]; then
echo "renaming $original_contigs to ${original_contigs}sta"
mv $original_contigs ${original_contigs}sta
original_contigs=${original_contigs}sta
fi
if [ ${original_contigs: -4} == ".fna" ]; then
echo "renaming $original_contigs to ${original_contigs%fna}fasta"
mv $original_contigs ${original_contigs%fna}fasta
original_contigs=${original_contigs%fna}fasta
fi
if [ ${original_contigs: -4} == ".fsa" ]; then
echo "renaming $original_contigs to ${original_contigs%fsa}fasta"
mv $original_contigs ${original_contigs%fsa}fasta
original_contigs=${original_contigs%fsa}fasta
fi
# Removing contigs under $circ_length_cutoff nts and detecting circular contigs
if [ $circ_length_cutoff -gt $linear_length_cutoff ] ; then
LENGTH_MINIMUM=$linear_length_cutoff
else
LENGTH_MINIMUM=$circ_length_cutoff
fi
if [ ${original_contigs: -6} == ".fasta" ]; then
echo "$(tput setaf 5)File with .fasta extension detected, attempting to keep contigs over $LENGTH_MINIMUM nt and find circular sequences with apc.pl$(tput sgr 0)"
bioawk -v run_var="$run_title" -v contig_cutoff="$LENGTH_MINIMUM" -c fastx '{ if(length($seq) > contig_cutoff) { print ">"run_var NR" "$name; print $seq }}' $original_contigs > ${original_contigs%.fasta}.over_${LENGTH_MINIMUM}nt.fasta ;
cd $run_title
echo "unlimited_breadsticks" > ${run_title}_CONTIG_SUMMARY.tsv
perl ${CENOTE_SCRIPT_DIR}/apc_cenote1.pl -b $run_title -c $CENOTE_SCRIPT_DIR ../${original_contigs%.fasta}.over_${LENGTH_MINIMUM}nt.fasta >/dev/null 2>&1
rm -f apc_aln*
APC_CIRCS=$( find . -maxdepth 1 -type f -name "${run_title}*.fa" | sed 's/\.\///g' )
if [ -n "$APC_CIRCS" ] ;then
for fa1 in $APC_CIRCS ; do
CIRC_SEQ_NAME=$( head -n1 $fa1 | sed 's/|.*//g' ) ;
CIRC_NEW_NAME=$( echo "$CIRC_SEQ_NAME" | sed 's/>//g ; s/ .*//g' )
grep -A1 "^$CIRC_SEQ_NAME" ../${original_contigs%.fasta}.over_${LENGTH_MINIMUM}nt.fasta | sed '/--/d' > ${CIRC_NEW_NAME}.fasta
rm -f $fa1
echo ${CIRC_NEW_NAME}.fasta" is circular/has DTRs"
done
else
echo "No circular contigs detected."
fi
elif [ ${original_contigs: -6} == ".fastg" ]; then
bioawk -v contig_cutoff="$LENGTH_MINIMUM" -c fastx '{ if(length($seq) > contig_cutoff) {print }}' $original_contigs | grep "[a-zA-Z0-9]:\|[a-zA-Z0-9];" | grep -v "':" | awk '{ print ">"$1 ; print $2 }' | sed 's/:.*//g; s/;.*//g' | bioawk -v run_var="$run_title" -c fastx '{ print ">"run_var NR" "$name; print $seq }' > ${original_contigs%.fastg}.over_${LENGTH_MINIMUM}nt.fasta
cd $run_title
echo "unlimited_breadsticks" > ${run_title}_CONTIG_SUMMARY.tsv
perl ${CENOTE_SCRIPT_DIR}/apc_cenote1.pl -b $run_title -c $CENOTE_SCRIPT_DIR ../${original_contigs%.fastg}.over_${LENGTH_MINIMUM}nt.fasta >/dev/null 2>&1
rm -f apc_aln*
APC_CIRCS=$( find . -maxdepth 1 * -type f -name "${run_title}*.fa" | sed 's/\.\///g' )
if [ -n "$APC_CIRCS" ] ;then
for fa1 in $APC_CIRCS ; do
CIRC_SEQ_NAME=$( head -n1 $fa1 | sed 's/|.*//g' ) ;
CIRC_NEW_NAME=$( echo "$CIRC_SEQ_NAME" | sed 's/>//g ; s/ .*//g' )
grep -A1 "^$CIRC_SEQ_NAME" ../${original_contigs%.fastg}.over_${LENGTH_MINIMUM}nt.fasta | sed '/--/d' > ${CIRC_NEW_NAME}.fasta
rm -f $fa1
echo ${CIRC_NEW_NAME}.fasta" is circular/has DTRs"
done
else
echo "No circular contigs detected."
fi
else
echo "$(tput setaf 4)File with .fasta of .fastg extension not detected as first input. Exiting.$(tput sgr 0)" ;
exit
fi
# Removing cirles that are smaller than user specified cutoff
CIRC_CONTIGS=$( find . -maxdepth 1 -type f -name "*.fasta" | sed 's/\.\///g' )
if [ ! -z "$CIRC_CONTIGS" ] ;then
for CIRCLE1 in $CIRC_CONTIGS ; do
CIRCLE1_LENGTH=$( bioawk -c fastx '{print length($seq) }' $CIRCLE1 )
if [[ $CIRCLE1_LENGTH -lt $circ_length_cutoff ]] ; then
mv $CIRCLE1 ${CIRCLE1%.fasta}.too_short.fasta
fi
done
fi
rm -f *.too_short.fasta
# Detecting whether any circular contigs were present
original_fastas=$( find . -maxdepth 1 -type f -name "*.fasta" | sed 's/\.\///g' )
# "$(tput setaf 5)$var1$(tput sgr 0)"
if [ -z "$original_fastas" ] ; then
echo "$(tput setaf 4)No circular fasta files detected. $(tput sgr 0)"
#exit
mkdir other_contigs
if [ ${original_contigs: -6} == ".fasta" ]; then
grep -A1 "^>" ../${original_contigs%.fasta}.over_${LENGTH_MINIMUM}nt.fasta | sed 's/--//g' | bioawk -v contig_cutoff="$linear_length_cutoff" -c fastx '{ if (length($seq) > contig_cutoff) { print ">"$name" "$4 ; print $seq }}' > other_contigs/all_non_circular.fasta
elif [ ${original_contigs: -6} == ".fastg" ]; then
grep -A1 "^>" ../${original_contigs%.fastg}.over_${LENGTH_MINIMUM}nt.fasta | sed 's/--//g' | bioawk -v contig_cutoff="$linear_length_cutoff" -c fastx '{ if (length($seq) > contig_cutoff) { print ">"$name" "$4 ; print $seq }}' > other_contigs/all_non_circular.fasta
fi
if [ -s other_contigs/all_non_circular.fasta ] ; then
grep "^>" other_contigs/all_non_circular.fasta | sed 's/>//g' | cut -d " " -f1 | while read LINE ; do
grep -A1 "$LINE [a-zA-Z]" other_contigs/all_non_circular.fasta > other_contigs/$LINE.fasta ;
done
fi
else
echo "$(tput setaf 5)Circular fasta file(s) detected$(tput sgr 0)"
echo " "
cat *fasta > all_circular_contigs_${run_title}.fna
# Putting non-circular contigs in a separate directory
echo "$(tput setaf 4)Putting non-circular contigs in a separate directory $(tput sgr 0)"
mkdir other_contigs
for CIRCLE in $original_fastas ; do
grep "^>" $CIRCLE | sed 's/|.*//g' >> circular_contigs_spades_names.txt
done
if [ ${original_contigs: -6} == ".fasta" ]; then
LINEAR_COUNT=$( grep -v -f circular_contigs_spades_names.txt ../${original_contigs%.fasta}.over_${LENGTH_MINIMUM}nt.fasta | grep -A1 "^>" | wc -l )
if [ $LINEAR_COUNT -ge 1 ] ; then
grep -v -f circular_contigs_spades_names.txt ../${original_contigs%.fasta}.over_${LENGTH_MINIMUM}nt.fasta | grep -A1 "^>" | sed 's/--//g' | bioawk -v contig_cutoff="$linear_length_cutoff" -c fastx '{ if (length($seq) > contig_cutoff) { print ">"$name" "$4 ; print $seq }}' > other_contigs/all_non_circular.fasta
else
echo "No linear contigs over ${LENGTH_MINIMUM}nt found in run"
fi
elif [ ${original_contigs: -6} == ".fastg" ]; then
LINEAR_COUNT=$( grep -v -f circular_contigs_spades_names.txt ../${original_contigs%.fastg}.over_${LENGTH_MINIMUM}nt.fasta | grep -A1 "^>" | wc -l )
if [ $LINEAR_COUNT -ge 1 ] ; then
grep -v -f circular_contigs_spades_names.txt ../${original_contigs%.fastg}.over_${LENGTH_MINIMUM}nt.fasta | grep -A1 "^>" | sed 's/--//g' | bioawk -v contig_cutoff="$linear_length_cutoff" -c fastx '{ if (length($seq) > contig_cutoff) { print ">"$name" "$4 ; print $seq }}' > other_contigs/all_non_circular.fasta
else
echo "No linear contigs over ${LENGTH_MINIMUM}nt found in run"
fi
fi
if [ -s other_contigs/all_non_circular.fasta ] ; then
grep "^>" other_contigs/all_non_circular.fasta | sed 's/>//g' | cut -d " " -f1 | while read LINE ; do
grep -A1 "$LINE [a-zA-Z0-9]" other_contigs/all_non_circular.fasta > other_contigs/$LINE.fasta ;
done
fi
fi
cd other_contigs
CONTIGS_NON_CIRCULAR=$( find . -maxdepth 1 -type f -name "*[0-9].fasta" | sed 's/\.\///g' )
if [ ! -z "$CONTIGS_NON_CIRCULAR" ] ;then
echo "$(tput setaf 4) Looking for non-circular contigs that have at least 1 virus-specific or plasmid-specific domain $(tput sgr 0)"
mkdir ../no_end_contigs_with_viral_domain
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: running prodigal on linear contigs " $MDYT
echo "$CONTIGS_NON_CIRCULAR" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t prodigal -a {}.AA.fasta -i {}.fasta -p meta -q >/dev/null 2>&1
for NO_END in $CONTIGS_NON_CIRCULAR ; do
sed 's/ /@/g' ${NO_END%.fasta}.AA.fasta | bioawk -c fastx '{print}' | while read LINE ; do
START_BASE=$( echo "$LINE" | cut -d "#" -f 2 | sed 's/@//g' ) ;
END_BASE=$( echo "$LINE" | cut -d "#" -f 3 | sed 's/@//g' ) ;
ORF_NAME=$( echo "$LINE" | cut -d "#" -f 1 | sed 's/@//g; s/\./_/g' ) ;
AA_SEQ=$( echo "$LINE" | cut -f2 | sed 's/\*//g' ) ;
echo ">"${ORF_NAME} "["$START_BASE" - "$END_BASE"]" ; echo $AA_SEQ ;
done > ${NO_END%.fasta}.AA.sorted.fasta
done
###instructions for large genomes
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: splitting running hmmscan for linear contigs against virus hallmark gene database: $virus_domain_db " $MDYT
cat $( find . -maxdepth 1 -type f -name "*.AA.sorted.fasta" ) > all_large_genome_proteins.AA.fasta
TOTAL_AA_SEQS=$( grep -F ">" all_large_genome_proteins.AA.fasta | wc -l | bc )
AA_SEQS_PER_FILE=$( echo "scale=0 ; $TOTAL_AA_SEQS / $CPU" | bc )
if [ $AA_SEQS_PER_FILE = 0 ] ; then
AA_SEQS_PER_FILE=1
fi
awk -v seq_per_file="$AA_SEQS_PER_FILE" 'BEGIN {n_seq=0;} /^>/ {if(n_seq%seq_per_file==0){file=sprintf("SPLIT_LARGE_GENOME_AA_%d.fasta",n_seq);} print >> file; n_seq++; next;} { print >> file; }' < all_large_genome_proteins.AA.fasta
SPLIT_AA_LARGE=$( find . -maxdepth 1 -type f -name "SPLIT_LARGE_GENOME_AA_*.fasta" | sed 's/\.\///g' )
if [[ $virus_domain_db = "standard" ]] ; then
echo "$SPLIT_AA_LARGE" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan.out --cpu 1 -E 1e-8 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/virus_specific_baits_plus_missed6a {}.fasta >/dev/null 2>&1
echo "$SPLIT_AA_LARGE" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan_replicate.out --cpu 1 -E 1e-15 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/virus_replication_clusters3 {}.fasta >/dev/null 2>&1
elif [[ $virus_domain_db = "virion" ]]; then
echo "$SPLIT_AA_LARGE" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan.out --cpu 1 -E 1e-8 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/virus_specific_baits_plus_missed6a {}.fasta >/dev/null 2>&1
elif [[ $virus_domain_db = "rna_virus" ]]; then
echo "$SPLIT_AA_LARGE" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan.out --cpu 1 -E 1e-8 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/rna_virus_rdrp_capsid_hmms1 {}.fasta >/dev/null 2>&1
else
echo "$(tput setaf 5) Incorrect argument given for virus_domain_db variable. Try standard, virion, or rna_virus as arguments. For this run, no contigs with viral domains but without circularity or ITRs will be detected $(tput sgr 0)"
rm -f ./*{0..9}.fasta
break
fi
HMM_REP_NUMEBR=$( find . -maxdepth 1 -type f -name "SPLIT_LARGE_GENOME_AA_*AA.hmmscan_replicate.out" | wc -l )
if [[ $FOR_PLASMIDS = "True" ]]; then
if [ $HMM_REP_NUMEBR -gt 0 ] ; then
cat SPLIT_LARGE_GENOME_AA_*AA.hmmscan.out SPLIT_LARGE_GENOME_AA_*AA.hmmscan_replicate.out | grep -v "^#\|plasmid_clust" | sed 's/ \+/ /g' | sort -u -k3,3 > LARGE_GENOME_COMBINED.AA.hmmscan.sort.out
else
cat SPLIT_LARGE_GENOME_AA_*AA.hmmscan.out | grep -v "^#\|plasmid_clust" | sed 's/ \+/ /g' | sort -u -k3,3 > LARGE_GENOME_COMBINED.AA.hmmscan.sort.out
fi
else
if [ $HMM_REP_NUMEBR -gt 0 ] ; then
cat SPLIT_LARGE_GENOME_AA_*AA.hmmscan.out SPLIT_LARGE_GENOME_AA_*AA.hmmscan_replicate.out | grep -v "^#" | sed 's/ \+/ /g' | sort -u -k3,3 > LARGE_GENOME_COMBINED.AA.hmmscan.sort.out
else
cat SPLIT_LARGE_GENOME_AA_*AA.hmmscan.out | grep -v "^#" | sed 's/ \+/ /g' | sort -u -k3,3 > LARGE_GENOME_COMBINED.AA.hmmscan.sort.out
fi
fi
if [ -s LARGE_GENOME_COMBINED.AA.hmmscan.sort.out ] ; then
cut -f3 LARGE_GENOME_COMBINED.AA.hmmscan.sort.out | sed 's/[^_]*$//' | sed 's/\(.*\)_/\1/' | sort -u | while read HIT ; do
HALL_COUNT=$( grep "${HIT}_" LARGE_GENOME_COMBINED.AA.hmmscan.sort.out | wc -l | bc )
if [ $HALL_COUNT -ge $LIN_MINIMUM_DOMAINS ] ; then
### not sure on this
mv ${HIT}.fasta ../no_end_contigs_with_viral_domain/${HIT}.fna
grep "${HIT}_" LARGE_GENOME_COMBINED.AA.hmmscan.sort.out > ../no_end_contigs_with_viral_domain/${HIT}.AA.hmmscan.sort.out
# ../no_end_contigs_with_viral_domain/${NO_END%.fasta}.no_hmmscan1.fasta
grep "${HIT}_" LARGE_GENOME_COMBINED.AA.hmmscan.sort.out | sort -u -k3,3 | cut -f3 | sed 's/\(.*\)/\1 /' > ${HIT}.AA.called_hmmscan.txt
grep -v -f ${HIT}.AA.called_hmmscan.txt ${HIT}.AA.sorted.fasta | grep -A1 ">" | sed '/--/d' > ../no_end_contigs_with_viral_domain/${HIT}.AA.no_hmmscan1.fasta
mv ${HIT}.AA.sorted.fasta ../no_end_contigs_with_viral_domain/
else
cat ${HIT}.fasta >> non_viral_domains_contigs.fna
rm -f ${HIT}.fasta
fi
done
fi
for REMAINDER in $CONTIGS_NON_CIRCULAR ; do
if [ -s $REMAINDER ] ; then
cat $REMAINDER >> non_viral_domains_contigs.fna
rm -f $REMAINDER
fi
done
fi
cd ..
CIRCLES_AND_ITRS=$( find . -maxdepth 1 -type f -regextype sed -regex "./${run_title}[0-9]\{1,6\}.fasta" | sed 's/\.\///g' )
# 3 ORF calling
if [ ! -z "$CIRCLES_AND_ITRS" ] ; then
mkdir DTR_contigs_with_viral_domain
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: Calling ORFs for circular/DTR sequences with prodigal " $MDYT
echo "$CIRCLES_AND_ITRS" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t prodigal -a {}.AA.fasta -i {}.fasta -p meta -q >/dev/null 2>&1
for CIRC in $CIRCLES_AND_ITRS ; do
sed 's/ /@/g' ${CIRC%.fasta}.AA.fasta | bioawk -c fastx '{print}' | while read LINE ; do
START_BASE=$( echo "$LINE" | cut -d "#" -f 2 | sed 's/@//g' ) ;
END_BASE=$( echo "$LINE" | cut -d "#" -f 3 | sed 's/@//g' ) ;
ORF_NAME=$( echo "$LINE" | cut -d "#" -f 1 | sed 's/@//g; s/\./_/g' ) ;
AA_SEQ=$( echo "$LINE" | cut -f2 | sed 's/\*//g' ) ;
echo ">"${ORF_NAME} "["$START_BASE" - "$END_BASE"]" ; echo $AA_SEQ ;
done > ${CIRC%.fasta}.AA.sorted.fasta
done
### stopping point -insert hmmscan
cat $( find . -maxdepth 1 -type f -name "*.AA.sorted.fasta" ) > all_circular_genome_proteins.AA.fasta
TOTAL_AA_SEQS=$( grep -F ">" all_circular_genome_proteins.AA.fasta | wc -l | bc )
AA_SEQS_PER_FILE=$( echo "scale=0 ; $TOTAL_AA_SEQS / $CPU" | bc )
if [ $AA_SEQS_PER_FILE = 0 ] ; then
AA_SEQS_PER_FILE=1
fi
awk -v seq_per_file="$AA_SEQS_PER_FILE" 'BEGIN {n_seq=0;} /^>/ {if(n_seq%seq_per_file==0){file=sprintf("SPLIT_CIRCULAR_AA_%d.fasta",n_seq);} print >> file; n_seq++; next;} { print >> file; }' < all_circular_genome_proteins.AA.fasta
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: running hmmscan on circular/DTR contigs on virus hallmark gene database: $virus_domain_db" $MDYT
SPLIT_AA_CIRC=$( find . -maxdepth 1 -type f -name "SPLIT_CIRCULAR_AA_*.fasta" | sed 's/\.\///g' )
if [[ $virus_domain_db = "standard" ]] ; then
echo "$SPLIT_AA_CIRC" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan.out --cpu 1 -E 1e-8 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/virus_specific_baits_plus_missed6a {}.fasta >/dev/null 2>&1
echo "$SPLIT_AA_CIRC" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan_replicate.out --cpu 1 -E 1e-15 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/virus_replication_clusters3 {}.fasta >/dev/null 2>&1
elif [[ $virus_domain_db = "virion" ]]; then
echo "$SPLIT_AA_CIRC" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan.out --cpu 1 -E 1e-8 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/virus_specific_baits_plus_missed6a {}.fasta >/dev/null 2>&1
elif [[ $virus_domain_db = "rna_virus" ]]; then
echo "$SPLIT_AA_CIRC" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan.out --cpu 1 -E 1e-8 --noali ${CENOTE_SCRIPT_DIR}/hmmscan_DBs/rna_virus_rdrp_capsid_hmms1 {}.fasta >/dev/null 2>&1
else
echo "$(tput setaf 5) Incorrect argument given for virus_domain_db variable. Try standard, virion, or rna_virus as arguments. For this run, no contigs with viral domains but without circularity or ITRs will be detected $(tput sgr 0)"
rm -f ./*{0..9}.fasta
break
fi
HMM_REP_NUMEBR=$( find . -maxdepth 1 -type f -name "SPLIT_CIRCULAR_AA_*AA.hmmscan_replicate.out" | wc -l )
if [[ $FOR_PLASMIDS = "True" ]]; then
if [ $HMM_REP_NUMEBR -gt 0 ] ; then
cat SPLIT_CIRCULAR_AA_*AA.hmmscan.out SPLIT_CIRCULAR_AA_*AA.hmmscan_replicate.out | grep -v "^#\|plasmid_clust" | sed 's/ \+/ /g' | sort -u -k3,3 > CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out
else
cat SPLIT_CIRCULAR_AA_*AA.hmmscan.out | grep -v "^#\|plasmid_clust" | sed 's/ \+/ /g' | sort -u -k3,3 > CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out
fi
else
if [ $HMM_REP_NUMEBR -gt 0 ] ; then
cat SPLIT_CIRCULAR_AA_*AA.hmmscan.out SPLIT_CIRCULAR_AA_*AA.hmmscan_replicate.out | grep -v "^#" | sed 's/ \+/ /g' | sort -u -k3,3 > CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out
else
cat SPLIT_CIRCULAR_AA_*AA.hmmscan.out | grep -v "^#" | sed 's/ \+/ /g' | sort -u -k3,3 > CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out
fi
fi
if [ -s CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out ] ; then
cut -f3 CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out | sed 's/[^_]*$//' | sed 's/\(.*\)_/\1/' | sort -u | while read HIT ; do
HALL_COUNT=$( grep "${HIT}_" CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out | wc -l | bc )
if [ $HALL_COUNT -ge $CIRC_MINIMUM_DOMAINS ] ; then
### not sure on this
mv ${HIT}.fasta DTR_contigs_with_viral_domain/${HIT}.fna
grep "${HIT}_" CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out > DTR_contigs_with_viral_domain/${HIT}.AA.hmmscan.sort.out
# ../no_end_contigs_with_viral_domain/${NO_END%.fasta}.no_hmmscan1.fasta
grep "${HIT}_" CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out | sort -u -k3,3 | cut -f3 | sed 's/\(.*\)/\1 /' > ${HIT}.AA.called_hmmscan.txt
grep -v -f ${HIT}.AA.called_hmmscan.txt ${HIT}.AA.sorted.fasta | grep -A1 ">" | sed '/--/d' > DTR_contigs_with_viral_domain/${HIT}.AA.no_hmmscan1.fasta
mv ${HIT}.AA.sorted.fasta DTR_contigs_with_viral_domain/
rm ${HIT}.AA.fasta
else
cat ${HIT}.fasta >> non_viral_domains_contigs.fna
rm -f ${HIT}.fasta
fi
done
fi
for REMAINDER in $CONTIGS_NON_CIRCULAR ; do
if [ -s $REMAINDER ] ; then
cat $REMAINDER >> other_contigs/non_viral_domains_contigs.fna
rm -f $REMAINDER
fi
done
fi
cd ${base_directory}/${run_title}
echo "ORIGINAL_NAME CENOTE_NAME END_FEATURE LENGTH NUM_HALLMARKS HALLMARK_NAMES" > ${run_title}_CONTIG_SUMMARY.tsv
CIRCULAR_HALLMARK_CONTIGS=$( find . -maxdepth 2 -type f -wholename "./DTR_contigs_with_viral_domain/*fna" | sed 's/\.\///g' )
if [ -n "$CIRCULAR_HALLMARK_CONTIGS" ] ; then
for LINEAR in $CIRCULAR_HALLMARK_CONTIGS ; do
CENOTE_NAME=$( head -n1 $LINEAR | cut -d " " -f1 | sed 's/>//g' )
ORIGINAL_NAME=$( head -n1 $LINEAR | cut -d " " -f2 )
LENGTH=$( bioawk -c fastx '{print length($seq)}' $LINEAR )
NUM_HALLMARKS=$( cat ${LINEAR%.fna}.AA.hmmscan.sort.out | wc -l | bc )
HALLMARK_NAMES=$( cut -f1 ${LINEAR%.fna}.AA.hmmscan.sort.out | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n/\|/g' | sort -u | sed 's/,//g' )
END_FEATURE="DTR"
echo "${ORIGINAL_NAME} ${CENOTE_NAME} ${END_FEATURE} ${LENGTH} ${NUM_HALLMARKS} ${HALLMARK_NAMES}" >> ${run_title}_CONTIG_SUMMARY.tsv
done
fi
if [ -s all_circular_genome_proteins.AA.fasta ] ; then
mv all_circular_genome_proteins.AA.fasta DTR_contigs_with_viral_domain/
fi
if [ -s CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out ] ; then
mv CIRCULAR_GENOME_COMBINED.AA.hmmscan.sort.out DTR_contigs_with_viral_domain/
fi
LIST_OF_VIRAL_DOMAIN_CONTIGS=$( find . -maxdepth 2 -type f -wholename "./no_end_contigs_with_viral_domain/*fna" | sed 's/\.\///g' )
if [ -n "$LIST_OF_VIRAL_DOMAIN_CONTIGS" ] ; then
for LINEAR in $LIST_OF_VIRAL_DOMAIN_CONTIGS ; do
CENOTE_NAME=$( head -n1 $LINEAR | cut -d " " -f1 | sed 's/>//g' )
ORIGINAL_NAME=$( head -n1 $LINEAR | cut -d " " -f2 )
LENGTH=$( bioawk -c fastx '{print length($seq)}' $LINEAR )
NUM_HALLMARKS=$( cat ${LINEAR%.fna}.AA.hmmscan.sort.out | wc -l | bc )
HALLMARK_NAMES=$( cut -f1 ${LINEAR%.fna}.AA.hmmscan.sort.out | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n/\|/g' | sort -u | sed 's/,//g' )
END_FEATURE="None"
echo "${ORIGINAL_NAME} ${CENOTE_NAME} ${END_FEATURE} ${LENGTH} ${NUM_HALLMARKS} ${HALLMARK_NAMES}" >> ${run_title}_CONTIG_SUMMARY.tsv
done
fi
### script for pruning no_end contigs with viral domains
if [ ! -z "$LIST_OF_VIRAL_DOMAIN_CONTIGS" ] && [ "$PROPHAGE" == "True" ] ;then
echo "$(tput setaf 3) Starting pruning of non-DTR/circular contigs with viral domains $(tput sgr 0)"
. ${CENOTE_SCRIPT_DIR}/prune_linear_contigs_0.1.sh
fi
cd ${base_directory}/${run_title}
### make combined virus seq file
if [ -n "$CIRCULAR_HALLMARK_CONTIGS" ] ; then
for CIRC in $CIRCULAR_HALLMARK_CONTIGS ; do
sed 's/ /#/g' $CIRC | bioawk -c fastx '{print ">"$name" DTR" ; print $seq}' | sed 's/#/ /g' >> final_combined_virus_sequences_${run_title}.fna
done
fi
if [ "$PROPHAGE" == "True" ] ;then
LINEAR_HALLMARK_CONTIGS=$( find . -maxdepth 2 -type f -regextype sed -regex ".*_vs[0-9]\{1,2\}.fna" | sed 's/\.\///g' )
if [ -n "$LINEAR_HALLMARK_CONTIGS" ] ; then
for LIN in $LINEAR_HALLMARK_CONTIGS ; do
ORIGINAL_NAME=$( head -n1 ${LIN%_vs[0-9][0-9].fna}.fna | cut -d " " -f2 )
sed 's/ /#/g' $LIN | bioawk -v ORI="$ORIGINAL_NAME" -c fastx '{print ">"$name" "ORI" no_end_feature" ; print $seq}' | sed 's/#/ /g' >> final_combined_virus_sequences_${run_title}.fna
done
fi
else
if [ -n "$LIST_OF_VIRAL_DOMAIN_CONTIGS" ] ; then
for LIN in $LIST_OF_VIRAL_DOMAIN_CONTIGS ; do
sed 's/ /#/g' $LIN | bioawk -c fastx '{print ">"$name" no_end_feature" ; print $seq}' | sed 's/#/ /g' >> final_combined_virus_sequences_${run_title}.fna
done
fi
fi
echo "removing ancillary files"
rm -f *.all_start_stop.txt *.bad_starts.txt *.comb.tbl *.comb2.tbl *.good_start_orfs.txt *.hypo_start_stop.txt *.nucl_orfs.fa *.remove_hypo.txt *.log *.promer.contigs_with_ends.fa *.promer.promer *.out.hhr *.starting_orf.txt *.out.hhr *.nucl_orfs.txt *.called_hmmscan.txt *.hmmscan_replicate.out *.hmmscan.out *.rotate.no_hmmscan.fasta *.starting_orf.1.fa *.phan.*fasta *used_positions.txt *.prodigal.for_prodigal.fa *.prodigal.gff *.trnascan-se2.txt *.for_blastp.txt *.for_hhpred.txt circular_contigs_spades_names.txt SPLIT_CIRCULAR_AA*fasta all_circular_contigs_${run_title}.fna ${run_title}*fasta
rm -rf bt2_indices/
rm -f other_contigs/*.AA.fasta other_contigs/*.AA.sorted.fasta other_contigs/*.out other_contigs/*.dat other_contigs/*called_hmmscan.txt other_contigs/SPLIT_LARGE_GENOME_AA_*fasta
rm -f no_end_contigs_with_viral_domain/*.called_hmmscan2.txt no_end_contigs_with_viral_domain/*.hmmscan2.out no_end_contigs_with_viral_domain/*all_hhpred_queries.AA.fasta no_end_contigs_with_viral_domain/*.all_start_stop.txt no_end_contigs_with_viral_domain/*.trnascan-se2.txt no_end_contigs_with_viral_domain/*.for_hhpred.txt no_end_contigs_with_viral_domain/*.for_blastp.txt no_end_contigs_with_viral_domain/*.HH.tbl no_end_contigs_with_viral_domain/*.hypo_start_stop.txt no_end_contigs_with_viral_domain/*.remove_hypo.txt no_end_contigs_with_viral_domain/*.rps_nohits.fasta no_end_contigs_with_viral_domain/*.tax_guide.blastx.tab no_end_contigs_with_viral_domain/*.tax_orf.fasta no_end_contigs_with_viral_domain/*.trans.fasta no_end_contigs_with_viral_domain/*.called_hmmscan*.txt no_end_contigs_with_viral_domain/*.no_hmmscan*.fasta no_end_contigs_with_viral_domain/*.comb*.tbl
echo " "
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: Finishing " $MDYT
if [ -s final_combined_virus_sequences_${run_title}.fna ] ; then
NUMBER_VIRUSES=$( grep -F ">" final_combined_virus_sequences_${run_title}.fna | wc -l | bc )
DTR_TOTAL=$( grep -F " DTR" final_combined_virus_sequences_${run_title}.fna | wc -l | bc )
NO_END_TOTAL=$( grep -F " no_end_feature" final_combined_virus_sequences_${run_title}.fna | wc -l | bc )
echo "$(tput setaf 3)Virus prediction summary:$(tput sgr 0)"
echo "$NUMBER_VIRUSES virus contigs were detected/predicted. $DTR_TOTAL contigs had DTRs/circularity. $NO_END_TOTAL were linear/had no end features"
fi
if [ -s ${run_title}_PRUNING_INFO_TABLE.tsv ] ; then
PRUNE_ATTEMPTS=$( awk '{OFS="\t"}{FS="\t"}{ if ($6=="True") {print}}' ${run_title}_PRUNING_INFO_TABLE.tsv | wc -l | bc )
PRUNE_REMOVE=$( awk '{OFS="\t"}{FS="\t"}{ if ($6=="True" && $7=="True") {print}}' ${run_title}_PRUNING_INFO_TABLE.tsv | wc -l | bc )
PRUNE_REMAIN=$( awk '{OFS="\t"}{FS="\t"}{ if ($6=="True" && $7=="False") {print}}' ${run_title}_PRUNING_INFO_TABLE.tsv | wc -l | bc )
echo "$(tput setaf 3)Prophage pruning summary:$(tput sgr 0)"
echo "$PRUNE_ATTEMPTS linear contigs > 10 kb were run through pruning module, and $PRUNE_REMOVE virus sub-contigs (putative prophages/proviruses) were extracted from these. $PRUNE_REMAIN virus contigs were kept intact."
fi
echo "$(tput setaf 3)output directory: "$run_title" $(tput sgr 0)"
echo "$(tput setaf 3) >>>>>>CENOTE UNLIMITED BREADSTICKS HAS FINISHED SERVING BREADSTICKS<<<<<< $(tput sgr 0)"