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prune_linear_contigs_0.1.sh
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prune_linear_contigs_0.1.sh
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#!/bin/bash
# Split from anotation_of_viral_domain_contigs.sh on 190919
cd no_end_contigs_with_viral_domain/
echo "pruning script opened"
vd_fastas=$( find . -maxdepth 1 -type f -name "*.fna" )
if [ -n "$vd_fastas" ] ; then
echo "fna files found"
for NO_END in $vd_fastas ; do
LENGTH_SEQ=$( bioawk -c fastx '{print length($seq)}' $NO_END )
if [[ "$LENGTH_SEQ" -lt 10000 ]] ; then
bioawk -c fastx '{print ">"$name"_vs01 "$4 ; print $seq}' $NO_END > ${NO_END%.fna}_vs01.fna
mv ${NO_END%.fna}.AA.sorted.fasta ${NO_END%.fna}_vs01.AA.sorted.fasta
echo "$NO_END is too short to prune chromosomal regions"
elif [[ "$LENGTH_SEQ" -gt 10000 ]] || [[ "$LENGTH_SEQ" == 10000 ]] ; then
mv ${NO_END%.fna}.AA.sorted.fasta ${NO_END%.fna}.AA.sorted1.fasta
MDYT=$( date +"%m-%d-%y---%T" )
#grep -v "^#\|plasmid_cluster" ${NO_END%.fna}.AA.hmmscan.out | sed 's/ \+/ /g' | sort -u -k3,3 > ${NO_END%.fna}.AA.hmmscan.sort.out
if [ ! -z "${NO_END%.fna}.AA.hmmscan.sort.out" ] ; then
cut -f3 ${NO_END%.fna}.AA.hmmscan.sort.out | awk '{ print $0" " }' > ${NO_END%.fna}.AA.called_hmmscan1.txt ;
cat ${NO_END%.fna}.AA.called_hmmscan1.txt | while read LINE ; do
PROTEIN_INFO=$( grep "$LINE \[" ${NO_END%.fna}.AA.sorted1.fasta ) ;
START_BASEH=$( echo $PROTEIN_INFO | sed 's/.*\[\(.*\) -.*/\1/' ) ;
END_BASEH=$( echo $PROTEIN_INFO | sed 's/.*- \(.*\)\].*/\1/' ) ;
HMM_INFO=$( grep "$LINE " ${NO_END%.fna}.AA.hmmscan.sort.out | head -n1 | cut -f1 | sed 's/-/ /g; s/.*[0-9]\+\///g' ) ;
INFERENCEH=$( echo $HMM_INFO | cut -d " " -f1 ) ;
echo -e "$LINE\t$START_BASEH\t$END_BASEH\t$INFERENCEH\t$HMM_INFO"
done > ${NO_END%.fna}.VIRUS_BAIT_TABLE.txt
fi
fi
done
### redo hmmscan parallelization
FOR_SCAN=$( find . -maxdepth 1 -type f -name "*.AA.sorted1.fasta" )
if [ -n "$FOR_SCAN" ] ; then
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: HMMSCAN of common viral domains beginning" $MDYT
#cat $( find . -maxdepth 1 -type f -name "*.AA.sorted1.fasta" ) > all_prunable_seq_proteins.AA.fasta
for SCANQ in $FOR_SCAN ; do
cat $SCANQ
done > all_prunable_seq_proteins.AA.fasta
TOTAL_AA_SEQS=$( grep -F ">" all_prunable_seq_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_PRUNE_SEQ_AA_%d.fasta",n_seq);} print >> file; n_seq++; next;} { print >> file; }' < all_prunable_seq_proteins.AA.fasta
SPLIT_AA_PRUNE=$( find . -maxdepth 1 -type f -name "SPLIT_PRUNE_SEQ_AA_*.fasta" )
echo "$SPLIT_AA_PRUNE" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t hmmscan --tblout {}.AA.hmmscan2.out --cpu 1 -E 1e-8 --noali ${CENOTE_DBS}/hmmscan_DBs/useful_hmms_baits_and_not2a {}.fasta >/dev/null 2>&1
cat SPLIT_PRUNE_SEQ_AA_*AA.hmmscan2.out | grep -v "^#" | sed 's/ \+/ /g' | sort -u -k3,3 > SPLIT_PRUNE_SEQ_COMBINED.AA.hmmscan2.sort.out
if [ -s SPLIT_PRUNE_SEQ_COMBINED.AA.hmmscan2.sort.out ] ; then
cut -f3 SPLIT_PRUNE_SEQ_COMBINED.AA.hmmscan2.sort.out | sed 's/[^_]*$//' | sed 's/\(.*\)_/\1/' | sort -u | while read HIT ; do
### not sure on this
grep "${HIT}_" SPLIT_PRUNE_SEQ_COMBINED.AA.hmmscan2.sort.out > ${HIT}.AA.hmmscan2.sort.out
# ../no_end_contigs_with_viral_domain/${NO_END%.fasta}.no_hmmscan1.fasta
grep "${HIT}_" SPLIT_PRUNE_SEQ_COMBINED.AA.hmmscan2.sort.out | sort -u -k3,3 | cut -f3 | sed 's/\(.*\)/\1 /' > ${HIT}.AA.called_hmmscan2.txt
grep -v -f ${HIT}.AA.called_hmmscan2.txt ${HIT}.AA.sorted1.fasta | grep -A1 ">" | sed '/--/d' > ${HIT}.AA.prune_no_hmmscan2.fasta
done
fi
CALLED_VIRAL=$( find . -maxdepth 1 -type f -name "*.AA.called_hmmscan2.txt" )
if [ -n "$CALLED_VIRAL" ] ; then
for NO_END in $CALLED_VIRAL ; do
cat $NO_END | sed 's/ $//g' | while read LINE ; do
PROTEIN_INFO=$( grep "$LINE \[" ${NO_END%.AA.called_hmmscan2.txt}.AA.sorted1.fasta ) ;
START_BASEH=$( echo $PROTEIN_INFO | sed 's/.*\[\(.*\) -.*/\1/' ) ;
END_BASEH=$( echo $PROTEIN_INFO | sed 's/.*- \(.*\)\].*/\1/' ) ;
HMM_INFO=$( grep "$LINE " ${NO_END%.AA.called_hmmscan2.txt}.AA.hmmscan2.sort.out | head -n1 | cut -f1 | sed 's/-/ /g; s/.*[0-9]\+\///g' ) ;
INFERENCEH=$( echo $HMM_INFO | cut -d " " -f1 ) ;
PROTEIN_NAME=$( echo $HMM_INFO | cut -d " " -f2- ) ;
if [[ $INFERENCEH == *"UniProtKB"* ]]; then
echo -e "$START_BASEH\t""$END_BASEH\t""CDS\n""\t\t\tprotein_id\t""lcl|""$LINE\n""\t\t\tproduct\t""$PROTEIN_NAME\n""\t\t\tinference\t""similar to AA sequence:$INFERENCEH" >> ${NO_END%.AA.called_hmmscan2.txt}.SCAN.tbl ;
else
echo -e "$START_BASEH\t""$END_BASEH\t""CDS\n""\t\t\tprotein_id\t""lcl|""$LINE\n""\t\t\tproduct\t""$PROTEIN_NAME\n""\t\t\tinference\t""protein motif:$INFERENCEH" >> ${NO_END%.AA.called_hmmscan2.txt}.SCAN.tbl ;
fi
done
done
fi
SCAN1_TBL=$( find . -maxdepth 1 -type f -name "*.SCAN.tbl" )
if [ -n "$SCAN1_TBL" ] ; then
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: making tables for hmmscan and rpsblast outputs " $MDYT
for feat_tbl2 in $SCAN1_TBL ; do
SCAN_TBL_LENGTH=$( cat $feat_tbl2 | wc -l )
if [[ "$SCAN_TBL_LENGTH" -gt 1 ]] ; then
#echo "making scan table for $feat_tbl2"
grep "^[0-9]" -A3 $feat_tbl2 | sed '/--/d' | sed 's/ /_/g' | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n //g' | while read LINE ; do
if echo $LINE | grep -q "CDS" ; then
GENOME=${feat_tbl2%.SCAN.tbl}
FEAT_TYPE=$( echo $LINE | cut -d " " -f3 )
FEAT_START=$( echo $LINE | cut -d " " -f1 )
FEAT_END=$( echo $LINE | cut -d " " -f2 )
FEAT_NAME=$( echo $LINE | cut -d " " -f7 )
FEAT_ATT=$( echo $LINE | cut -d " " -f9 )
FEAT_ID=$( echo $LINE | cut -d " " -f5 | sed 's/lcl|//g' )
echo -e "$FEAT_ID\t$FEAT_START\t$FEAT_END\tVIRUS_COMMON\t$FEAT_NAME"
fi
done > ${feat_tbl2%.SCAN.tbl}.HMMSCAN_TABLE.txt
else
echo "not making scan table for $feat_tbl2"
fi
done
fi
### redo RPS part
NO_HMMSCAN_AA=$( find . -maxdepth 1 -type f -name "*.AA.prune_no_hmmscan2.fasta" )
if [ -n "$NO_HMMSCAN_AA" ] ; then
#cat $( find . -maxdepth 1 -type f -name "*.AA.prune_no_hmmscan2.fasta" ) > all_prunable_rps_proteins.AA.fasta
for NO_HMMQ in $NO_HMMSCAN_AA ; do
cat $NO_HMMQ
done > all_prunable_rps_proteins.AA.fasta
TOTAL_AA_SEQS=$( grep -F ">" all_prunable_rps_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_PRUNE_RPS_AA_%d.fasta",n_seq);} print >> file; n_seq++; next;} { print >> file; }' < all_prunable_rps_proteins.AA.fasta
SPLIT_AA_RPS=$( find . -maxdepth 1 -type f -name "SPLIT_PRUNE_RPS_AA_*.fasta" )
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: running RPSBLAST on each sequence " $MDYT
echo "$SPLIT_AA_RPS" | sed 's/.fasta//g' | xargs -n 1 -I {} -P $CPU -t rpsblast -evalue 1e-4 -num_descriptions 5 -num_alignments 1 -db ${CENOTE_DBS}/cdd_rps_db/Cdd -seg yes -query {}.fasta -line_length 200 -out {}.rpsb.out >/dev/null 2>&1
cat *rpsb.out > COMBINED_RESULTS_PRUNE.AA.rpsblast.out
perl ${CENOTE_SCRIPT_DIR}/rpsblastreport_to_table2.pl
cut -f1 COMBINED_RESULTS_PRUNE.RPS_TABLE.txt | sed 's/[^_]*$//' | sed 's/\(.*\)_/\1/' | sort -u | while read CONTIG ; do
grep "^${CONTIG}_" COMBINED_RESULTS_PRUNE.RPS_TABLE.txt > ${CONTIG}.RPS_TABLE.txt
done
fi
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: parsing tables into virus_signal.seq files for hmmscan and rpsblast outputs " $MDYT
HMM_TBL=$( find . -maxdepth 1 -type f -name "*.HMMSCAN_TABLE.txt" )
if [ -n "$HMM_TBL" ] ; then
for TABLE1 in $HMM_TBL ; do
#echo $TABLE1
CONTIG_LENGTH=$( bioawk -c fastx '{ print length($seq) }' ${TABLE1%.HMMSCAN_TABLE.txt}.fna )
awk -v lengthq="$CONTIG_LENGTH" 'BEGIN {for (i=1;i<=lengthq;i++) print i, "Z"}' | sed 's/ / /g' > ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
cat ${TABLE1%.HMMSCAN_TABLE.txt}.RPS_TABLE.txt | while read LINE ; do
RPS_START=$( echo "$LINE" | cut -f2 ) ;
RPS_END=$( echo "$LINE" | cut -f3 ) ;
INFER=$( echo "$LINE" | cut -f4 ) ;
if [ $INFER == "HYPO" ] ; then
if [[ "$RPS_END" -gt "$RPS_START" ]] ; then
awk -v startq="$RPS_START" -v endq="$RPS_END" 'BEGIN {for (i=startq;i<=endq;i++) print i, "X"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
elif [[ "$RPS_START" -gt "$RPS_END" ]] ; then
awk -v startq="$RPS_START" -v endq="$RPS_END" 'BEGIN {for (i=endq;i<=startq;i++) print i, "X"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
fi
elif [[ $INFER == *"PHA0"* ]] || grep -q "$INFER" ${CENOTE_SCRIPT_DIR}/viral_cdds_and_pfams_191028.txt ; then
if [[ "$RPS_END" -gt "$RPS_START" ]] ; then
awk -v startq="$RPS_START" -v endq="$RPS_END" 'BEGIN {for (i=startq;i<=endq;i++) print i, "V"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
elif [[ "$RPS_START" -gt "$RPS_END" ]] ; then
awk -v startq="$RPS_START" -v endq="$RPS_END" 'BEGIN {for (i=endq;i<=startq;i++) print i, "V"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
fi
else
if [[ "$RPS_END" -gt "$RPS_START" ]] ; then
awk -v startq="$RPS_START" -v endq="$RPS_END" 'BEGIN {for (i=startq;i<=endq;i++) print i, "Y"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
elif [[ "$RPS_START" -gt "$RPS_END" ]] ; then
awk -v startq="$RPS_START" -v endq="$RPS_END" 'BEGIN {for (i=endq;i<=startq;i++) print i, "Y"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
fi
fi
done
cat $TABLE1 | while read LINE ; do
VIR_START=$( echo "$LINE" | cut -f2 ) ;
VIR_END=$( echo "$LINE" | cut -f3 ) ;
if [[ "$VIR_END" -gt "$VIR_START" ]] ; then
awk -v startq="$VIR_START" -v endq="$VIR_END" 'BEGIN {for (i=startq;i<=endq;i++) print i, "V"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
elif [[ "$VIR_START" -gt "$VIR_END" ]] ; then
awk -v startq="$VIR_START" -v endq="$VIR_END" 'BEGIN {for (i=endq;i<=startq;i++) print i, "V"}' | sed 's/ / /g' >> ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab
fi ;
done
sort -g -k1,1 ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.tab | awk '!_[$1]++' | cut -f2 | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n//g' > ${TABLE1%.HMMSCAN_TABLE.txt}.virus_signal.seq
done
fi
else
echo "no contigs over 10kb (only sequences 10kb or greater can be pruned by this script)"
fi
else
echo "fna files NOT found"
fi
# Run Anna's python for extracting virus segments from potential prophage
if [ $PROPHAGE == "False" ] ; then
echo "prophages not being pruned"
else
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: Identifying virus chunks, chromosomal junctions, and pruning contigs as necessary " $MDYT
SIGNAL_SEQ=$( find . -maxdepth 1 -type f -name "*virus_signal.seq" )
if [ -n "$SIGNAL_SEQ" ] ; then
for VSEQ in *virus_signal.seq ; do
python ${CENOTE_SCRIPT_DIR}/cenote_virus_segments_V6.py $VSEQ
done
else
echo "No prophage signal files found"
fi
# reformat .fastas to get viral segments of prophage
CHUNK_COORDINATES=$( find . -maxdepth 1 -type f -name "*.virus_signal.seq_chunk_coordinates.csv" )
if [ -n "$CHUNK_COORDINATES" ] ; then
VIR_COUNTER=0
for CHUNKS in *.virus_signal.seq_chunk_coordinates.csv ; do
cat $CHUNKS | while read LINE ; do
CHUNK_START=$( echo "$LINE" | cut -d "," -f2 )
CHUNK_END=$( echo "$LINE" | cut -d "," -f3 )
CHUNK_VIR=$( cat ${CHUNKS%.virus_signal.seq_chunk_coordinates.csv}.VIRUS_BAIT_TABLE.txt | while read BAIT_LINE ; do echo "$BAIT_LINE" | awk -v CHUNK_STARTQ="$CHUNK_START" -v CHUNK_ENDQ="$CHUNK_END" '{ if ($3-(($3-$2)/2) >= CHUNK_STARTQ && $3-(($3-$2)/2) <= CHUNK_ENDQ) { print } }' ; done | wc -l ) ;
if [[ $CHUNK_VIR -gt $LIN_MINIMUM_DOMAINS ]] || [ $CHUNK_VIR == $LIN_MINIMUM_DOMAINS ] ; then
let VIR_COUNTER=VIR_COUNTER+1 ;
VIR_VALUE=$( printf "%02d" $VIR_COUNTER)
CHUNK_LENGTH=$(( ${CHUNK_END}-${CHUNK_START} ))
bioawk -v chunk_startq="$CHUNK_START" -v chunk_endq="$CHUNK_END" -v chunk_lengthq="$CHUNK_LENGTH" -v parent="${CHUNKS%.virus_signal.seq_chunk_coordinates.csv}" -v good_chunk="$VIR_VALUE" -c fastx '{ print ">"parent"_vs"good_chunk" "chunk_startq"-"chunk_endq ; print substr($seq, chunk_startq, chunk_lengthq)}' ${CHUNKS%.virus_signal.seq_chunk_coordinates.csv}.fna > ${CHUNKS%.virus_signal.seq_chunk_coordinates.csv}_vs${VIR_VALUE}.fna
fi
done
done
fi
fi
POST_PRUNE_CONTIGS=$( find . -maxdepth 1 -type f -regextype sed -regex ".*_vs[0-9]\{1,2\}.fna"| sed 's/\.\///g' )
if [ -n "$POST_PRUNE_CONTIGS" ] ; then
MDYT=$( date +"%m-%d-%y---%T" )
echo "time update: Making prophage table " $MDYT
echo "SEQUENCE_FILE CENOTE_PARENT_NAME INPUT_PARENT_NAME FINAL_LENGTH PARENT_LENGTH PRUNING_TRIED CHROM_REMOVED LEFT_JUNCTION RIGHT_JUNCTION HALLMARK_COUNT HALLMARK_GENES HALLMARK_AA_NAMES" > ${run_title}_PRUNING_INFO_TABLE.tsv
for CONTIG in $POST_PRUNE_CONTIGS ; do
CENOTE_PARENT=${CONTIG%_vs[0-9][0-9].fna}.fna
PARENT_LENGTH=$( bioawk -c fastx '{print length($seq)}' $CENOTE_PARENT )
PRUNED_LENGTH=$( bioawk -c fastx '{print length($seq)}' $CONTIG )
if [ -s ${CONTIG%_vs[0-9][0-9].fna}.VIRUS_BAIT_TABLE.txt ] ; then
LEFT_COORD=$( head -n1 $CONTIG | cut -d " " -f2 | cut -d "-" -f1 )
RIGHT_COORD=$( head -n1 $CONTIG | cut -d " " -f2 | cut -d "-" -f2 )
PRUNING_TRIED="True"
PQ_LENGTH=$(( ${PARENT_LENGTH}+1 ))
if [ $LEFT_COORD == 0 ] && [ $RIGHT_COORD == $PQ_LENGTH ] ; then
CHROM_REMOVED="False"
else
CHROM_REMOVED="True"
fi
HALLMARK_COUNT=$( awk -v LEFTQ="$LEFT_COORD" -v RIGHTQ="$RIGHT_COORD" '{FS="\t"}{OFS="\t"}{ if ($2>LEFTQ && $2<RIGHTQ && $3>LEFTQ && $3<RIGHTQ) {print $5}}' ${CONTIG%_vs[0-9][0-9].fna}.VIRUS_BAIT_TABLE.txt | wc -l | bc )
HALLMARK_GENES=$( awk -v LEFTQ="$LEFT_COORD" -v RIGHTQ="$RIGHT_COORD" '{FS="\t"}{OFS="\t"}{ if ($2>LEFTQ && $2<RIGHTQ && $3>LEFTQ && $3<RIGHTQ) {print $5}}' ${CONTIG%_vs[0-9][0-9].fna}.VIRUS_BAIT_TABLE.txt | sed 's/ /_/g ; s/,//g' | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n/\|/g' )
HALLMARK_AA_NAMES=$( awk -v LEFTQ="$LEFT_COORD" -v RIGHTQ="$RIGHT_COORD" '{ if ($2>LEFTQ && $2<RIGHTQ && $3>LEFTQ && $3<RIGHTQ) {print $1}}' ${CONTIG%_vs[0-9][0-9].fna}.VIRUS_BAIT_TABLE.txt | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n/\|/g' )
else
LEFT_COORD="None"
RIGHT_COORD="None"
PRUNING_TRIED="False"
CHROM_REMOVED="False"
HALLMARK_COUNT=$( cat ${CONTIG%_vs[0-9][0-9].fna}.AA.hmmscan.sort.out | wc -l | bc )
HALLMARK_GENES=$( cut -f1 ${CONTIG%_vs[0-9][0-9].fna}.AA.hmmscan.sort.out | sed 's/ /_/g ; s/,//g' | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n/\|/g' )
HALLMARK_AA_NAMES=$( cut -f3 ${CONTIG%_vs[0-9][0-9].fna}.AA.hmmscan.sort.out | sed 's/ /_/g ; s/,//g' | sed -e ':a' -e 'N' -e '$!ba' -e 's/\n/\|/g' )
fi
ORIGINAL_NAME=$( head -n1 ${CONTIG%_vs[0-9][0-9].fna}.fna | cut -d " " -f2 )
echo "${CONTIG} ${CENOTE_PARENT} ${ORIGINAL_NAME} ${PRUNED_LENGTH} ${PARENT_LENGTH} ${PRUNING_TRIED} ${CHROM_REMOVED} ${LEFT_COORD} ${RIGHT_COORD} ${HALLMARK_COUNT} ${HALLMARK_GENES} ${HALLMARK_AA_NAMES}" >> ${run_title}_PRUNING_INFO_TABLE.tsv
### finish this
done
fi
if [ -s ${run_title}_PRUNING_INFO_TABLE.tsv ] ; then
mv ${run_title}_PRUNING_INFO_TABLE.tsv ../
fi
rm -f *.virus_signal.tab *.used_positions.txt *.phan.fasta *.phan.sort.fasta *rpsb.out SPLIT_PRUNE_RPS_AA*fasta SPLIT_PRUNE_SEQ_AA*fasta
cd ..
echo "$(tput setaf 3) FINISHED PRUNING CONTIGS WITH AT LEAST $LIN_MINIMUM_DOMAINS VIRAL DOMAIN(S) $(tput sgr 0)"