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fix: move primer file location under workflow; this avoids file not b…
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…eing found in shared filesystems (#665)

* fix: move nCoV-2019.primer.bed under workflow/resources

* fix: fix path of primers in main config

* fix: fix primer path to be called internally

* fix: fix same in previous commit in read_clipping.smk

* define a function for getting kracken input in common.smk

* fix get_kracken_input()

* use get_kracken_input in qc.smk

* restore all changes

* refactor: move checking the primer file for URL into the script

* chore: fmt

* fix: disable formatting temporarily

* Update main.yml

* Update main.yml

* fmt script

* add missing key to artifact name

---------

Co-authored-by: Johannes Köster <[email protected]>
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@huzuner @johanneskoester
huzuner and johanneskoester authored Oct 23, 2024
1 parent 576329c commit 9cb1576
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Showing 8 changed files with 273 additions and 27 deletions.
6 changes: 2 additions & 4 deletions .github/workflows/main.yml
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Original file line number Diff line number Diff line change
Expand Up @@ -24,7 +24,7 @@ jobs:
steps:
- uses: actions/checkout@v4
with:
depth: 0
fetch-depth: 0
- name: Formatting
uses: super-linter/[email protected]
env:
Expand Down Expand Up @@ -66,7 +66,6 @@ jobs:
env:
GISAID_API_TOKEN: ${{ secrets.GISAID_API_TOKEN }}
needs:
- Formatting
- Linting
#- Pre-Commit
strategy:
Expand Down Expand Up @@ -181,7 +180,7 @@ jobs:
uses: actions/upload-artifact@v4
if: matrix.technology == 'all' && matrix.rule != 'all -npr'
with:
name: log-rule-${{ matrix.rule }}-technology-${{ matrix.technology }}
name: log-rule-${{ matrix.rule }}-technology-${{ matrix.technology }}-${{ matrix.seq_method }}
path: .tests/logs/

- name: Change permissions for caching
Expand All @@ -195,7 +194,6 @@ jobs:
env:
GISAID_API_TOKEN: ${{ secrets.GISAID_API_TOKEN }}
needs:
- Formatting
- Linting
#- Pre-Commit
strategy:
Expand Down
218 changes: 218 additions & 0 deletions workflow/resources/nCoV-2019.primer.bed
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@@ -0,0 +1,218 @@
MN908947.3 30 54 nCoV-2019_1_LEFT 1 +
MN908947.3 385 410 nCoV-2019_1_RIGHT 1 -
MN908947.3 320 342 nCoV-2019_2_LEFT 2 +
MN908947.3 704 726 nCoV-2019_2_RIGHT 2 -
MN908947.3 642 664 nCoV-2019_3_LEFT 1 +
MN908947.3 1004 1028 nCoV-2019_3_RIGHT 1 -
MN908947.3 943 965 nCoV-2019_4_LEFT 2 +
MN908947.3 1312 1337 nCoV-2019_4_RIGHT 2 -
MN908947.3 1242 1264 nCoV-2019_5_LEFT 1 +
MN908947.3 1623 1651 nCoV-2019_5_RIGHT 1 -
MN908947.3 1573 1595 nCoV-2019_6_LEFT 2 +
MN908947.3 1942 1964 nCoV-2019_6_RIGHT 2 -
MN908947.3 1875 1897 nCoV-2019_7_LEFT 1 +
MN908947.3 1868 1890 nCoV-2019_7_LEFT_alt0 1 +
MN908947.3 2247 2269 nCoV-2019_7_RIGHT 1 -
MN908947.3 2242 2264 nCoV-2019_7_RIGHT_alt5 1 -
MN908947.3 2181 2205 nCoV-2019_8_LEFT 2 +
MN908947.3 2568 2592 nCoV-2019_8_RIGHT 2 -
MN908947.3 2505 2529 nCoV-2019_9_LEFT 1 +
MN908947.3 2504 2528 nCoV-2019_9_LEFT_alt4 1 +
MN908947.3 2882 2904 nCoV-2019_9_RIGHT 1 -
MN908947.3 2880 2902 nCoV-2019_9_RIGHT_alt2 1 -
MN908947.3 2826 2850 nCoV-2019_10_LEFT 2 +
MN908947.3 3183 3210 nCoV-2019_10_RIGHT 2 -
MN908947.3 3144 3166 nCoV-2019_11_LEFT 1 +
MN908947.3 3507 3531 nCoV-2019_11_RIGHT 1 -
MN908947.3 3460 3482 nCoV-2019_12_LEFT 2 +
MN908947.3 3826 3853 nCoV-2019_12_RIGHT 2 -
MN908947.3 3771 3795 nCoV-2019_13_LEFT 1 +
MN908947.3 4142 4164 nCoV-2019_13_RIGHT 1 -
MN908947.3 4054 4077 nCoV-2019_14_LEFT 2 +
MN908947.3 4044 4068 nCoV-2019_14_LEFT_alt4 2 +
MN908947.3 4428 4450 nCoV-2019_14_RIGHT 2 -
MN908947.3 4402 4424 nCoV-2019_14_RIGHT_alt2 2 -
MN908947.3 4294 4321 nCoV-2019_15_LEFT 1 +
MN908947.3 4296 4322 nCoV-2019_15_LEFT_alt1 1 +
MN908947.3 4674 4696 nCoV-2019_15_RIGHT 1 -
MN908947.3 4666 4689 nCoV-2019_15_RIGHT_alt3 1 -
MN908947.3 4636 4658 nCoV-2019_16_LEFT 2 +
MN908947.3 4995 5017 nCoV-2019_16_RIGHT 2 -
MN908947.3 4939 4966 nCoV-2019_17_LEFT 1 +
MN908947.3 5296 5321 nCoV-2019_17_RIGHT 1 -
MN908947.3 5230 5259 nCoV-2019_18_LEFT 2 +
MN908947.3 5257 5287 nCoV-2019_18_LEFT_alt2 2 +
MN908947.3 5620 5644 nCoV-2019_18_RIGHT 2 -
MN908947.3 5620 5643 nCoV-2019_18_RIGHT_alt1 2 -
MN908947.3 5563 5586 nCoV-2019_19_LEFT 1 +
MN908947.3 5932 5957 nCoV-2019_19_RIGHT 1 -
MN908947.3 5867 5894 nCoV-2019_20_LEFT 2 +
MN908947.3 6247 6272 nCoV-2019_20_RIGHT 2 -
MN908947.3 6167 6196 nCoV-2019_21_LEFT 1 +
MN908947.3 6168 6197 nCoV-2019_21_LEFT_alt2 1 +
MN908947.3 6528 6550 nCoV-2019_21_RIGHT 1 -
MN908947.3 6526 6548 nCoV-2019_21_RIGHT_alt0 1 -
MN908947.3 6466 6495 nCoV-2019_22_LEFT 2 +
MN908947.3 6846 6873 nCoV-2019_22_RIGHT 2 -
MN908947.3 6718 6745 nCoV-2019_23_LEFT 1 +
MN908947.3 7092 7117 nCoV-2019_23_RIGHT 1 -
MN908947.3 7035 7058 nCoV-2019_24_LEFT 2 +
MN908947.3 7389 7415 nCoV-2019_24_RIGHT 2 -
MN908947.3 7305 7332 nCoV-2019_25_LEFT 1 +
MN908947.3 7671 7694 nCoV-2019_25_RIGHT 1 -
MN908947.3 7626 7651 nCoV-2019_26_LEFT 2 +
MN908947.3 7997 8019 nCoV-2019_26_RIGHT 2 -
MN908947.3 7943 7968 nCoV-2019_27_LEFT 1 +
MN908947.3 8319 8341 nCoV-2019_27_RIGHT 1 -
MN908947.3 8249 8275 nCoV-2019_28_LEFT 2 +
MN908947.3 8635 8661 nCoV-2019_28_RIGHT 2 -
MN908947.3 8595 8619 nCoV-2019_29_LEFT 1 +
MN908947.3 8954 8983 nCoV-2019_29_RIGHT 1 -
MN908947.3 8888 8913 nCoV-2019_30_LEFT 2 +
MN908947.3 9245 9271 nCoV-2019_30_RIGHT 2 -
MN908947.3 9204 9226 nCoV-2019_31_LEFT 1 +
MN908947.3 9557 9585 nCoV-2019_31_RIGHT 1 -
MN908947.3 9477 9502 nCoV-2019_32_LEFT 2 +
MN908947.3 9834 9858 nCoV-2019_32_RIGHT 2 -
MN908947.3 9784 9806 nCoV-2019_33_LEFT 1 +
MN908947.3 10146 10171 nCoV-2019_33_RIGHT 1 -
MN908947.3 10076 10099 nCoV-2019_34_LEFT 2 +
MN908947.3 10437 10459 nCoV-2019_34_RIGHT 2 -
MN908947.3 10362 10384 nCoV-2019_35_LEFT 1 +
MN908947.3 10737 10763 nCoV-2019_35_RIGHT 1 -
MN908947.3 10666 10688 nCoV-2019_36_LEFT 2 +
MN908947.3 11048 11074 nCoV-2019_36_RIGHT 2 -
MN908947.3 10999 11022 nCoV-2019_37_LEFT 1 +
MN908947.3 11372 11394 nCoV-2019_37_RIGHT 1 -
MN908947.3 11306 11331 nCoV-2019_38_LEFT 2 +
MN908947.3 11668 11693 nCoV-2019_38_RIGHT 2 -
MN908947.3 11555 11584 nCoV-2019_39_LEFT 1 +
MN908947.3 11927 11949 nCoV-2019_39_RIGHT 1 -
MN908947.3 11863 11889 nCoV-2019_40_LEFT 2 +
MN908947.3 12234 12256 nCoV-2019_40_RIGHT 2 -
MN908947.3 12110 12133 nCoV-2019_41_LEFT 1 +
MN908947.3 12465 12490 nCoV-2019_41_RIGHT 1 -
MN908947.3 12417 12439 nCoV-2019_42_LEFT 2 +
MN908947.3 12779 12802 nCoV-2019_42_RIGHT 2 -
MN908947.3 12710 12732 nCoV-2019_43_LEFT 1 +
MN908947.3 13074 13096 nCoV-2019_43_RIGHT 1 -
MN908947.3 13005 13027 nCoV-2019_44_LEFT 2 +
MN908947.3 13007 13029 nCoV-2019_44_LEFT_alt3 2 +
MN908947.3 13378 13400 nCoV-2019_44_RIGHT 2 -
MN908947.3 13363 13385 nCoV-2019_44_RIGHT_alt0 2 -
MN908947.3 13319 13344 nCoV-2019_45_LEFT 1 +
MN908947.3 13307 13336 nCoV-2019_45_LEFT_alt2 1 +
MN908947.3 13669 13699 nCoV-2019_45_RIGHT 1 -
MN908947.3 13660 13689 nCoV-2019_45_RIGHT_alt7 1 -
MN908947.3 13599 13621 nCoV-2019_46_LEFT 2 +
MN908947.3 13602 13625 nCoV-2019_46_LEFT_alt1 2 +
MN908947.3 13962 13984 nCoV-2019_46_RIGHT 2 -
MN908947.3 13961 13984 nCoV-2019_46_RIGHT_alt2 2 -
MN908947.3 13918 13946 nCoV-2019_47_LEFT 1 +
MN908947.3 14271 14299 nCoV-2019_47_RIGHT 1 -
MN908947.3 14207 14232 nCoV-2019_48_LEFT 2 +
MN908947.3 14579 14601 nCoV-2019_48_RIGHT 2 -
MN908947.3 14545 14570 nCoV-2019_49_LEFT 1 +
MN908947.3 14898 14926 nCoV-2019_49_RIGHT 1 -
MN908947.3 14865 14895 nCoV-2019_50_LEFT 2 +
MN908947.3 15224 15246 nCoV-2019_50_RIGHT 2 -
MN908947.3 15171 15193 nCoV-2019_51_LEFT 1 +
MN908947.3 15538 15560 nCoV-2019_51_RIGHT 1 -
MN908947.3 15481 15503 nCoV-2019_52_LEFT 2 +
MN908947.3 15861 15886 nCoV-2019_52_RIGHT 2 -
MN908947.3 15827 15851 nCoV-2019_53_LEFT 1 +
MN908947.3 16186 16209 nCoV-2019_53_RIGHT 1 -
MN908947.3 16118 16144 nCoV-2019_54_LEFT 2 +
MN908947.3 16485 16510 nCoV-2019_54_RIGHT 2 -
MN908947.3 16416 16444 nCoV-2019_55_LEFT 1 +
MN908947.3 16804 16833 nCoV-2019_55_RIGHT 1 -
MN908947.3 16748 16770 nCoV-2019_56_LEFT 2 +
MN908947.3 17130 17152 nCoV-2019_56_RIGHT 2 -
MN908947.3 17065 17087 nCoV-2019_57_LEFT 1 +
MN908947.3 17430 17452 nCoV-2019_57_RIGHT 1 -
MN908947.3 17381 17406 nCoV-2019_58_LEFT 2 +
MN908947.3 17738 17761 nCoV-2019_58_RIGHT 2 -
MN908947.3 17674 17697 nCoV-2019_59_LEFT 1 +
MN908947.3 18036 18062 nCoV-2019_59_RIGHT 1 -
MN908947.3 17966 17993 nCoV-2019_60_LEFT 2 +
MN908947.3 18324 18348 nCoV-2019_60_RIGHT 2 -
MN908947.3 18253 18275 nCoV-2019_61_LEFT 1 +
MN908947.3 18650 18672 nCoV-2019_61_RIGHT 1 -
MN908947.3 18596 18618 nCoV-2019_62_LEFT 2 +
MN908947.3 18957 18979 nCoV-2019_62_RIGHT 2 -
MN908947.3 18896 18918 nCoV-2019_63_LEFT 1 +
MN908947.3 19275 19297 nCoV-2019_63_RIGHT 1 -
MN908947.3 19204 19232 nCoV-2019_64_LEFT 2 +
MN908947.3 19591 19616 nCoV-2019_64_RIGHT 2 -
MN908947.3 19548 19570 nCoV-2019_65_LEFT 1 +
MN908947.3 19911 19939 nCoV-2019_65_RIGHT 1 -
MN908947.3 19844 19866 nCoV-2019_66_LEFT 2 +
MN908947.3 20231 20255 nCoV-2019_66_RIGHT 2 -
MN908947.3 20172 20200 nCoV-2019_67_LEFT 1 +
MN908947.3 20542 20572 nCoV-2019_67_RIGHT 1 -
MN908947.3 20472 20496 nCoV-2019_68_LEFT 2 +
MN908947.3 20867 20890 nCoV-2019_68_RIGHT 2 -
MN908947.3 20786 20813 nCoV-2019_69_LEFT 1 +
MN908947.3 21146 21169 nCoV-2019_69_RIGHT 1 -
MN908947.3 21075 21104 nCoV-2019_70_LEFT 2 +
MN908947.3 21427 21455 nCoV-2019_70_RIGHT 2 -
MN908947.3 21357 21386 nCoV-2019_71_LEFT 1 +
MN908947.3 21716 21743 nCoV-2019_71_RIGHT 1 -
MN908947.3 21658 21682 nCoV-2019_72_LEFT 2 +
MN908947.3 22013 22038 nCoV-2019_72_RIGHT 2 -
MN908947.3 21961 21990 nCoV-2019_73_LEFT 1 +
MN908947.3 22324 22346 nCoV-2019_73_RIGHT 1 -
MN908947.3 22262 22290 nCoV-2019_74_LEFT 2 +
MN908947.3 22626 22650 nCoV-2019_74_RIGHT 2 -
MN908947.3 22516 22542 nCoV-2019_75_LEFT 1 +
MN908947.3 22877 22903 nCoV-2019_75_RIGHT 1 -
MN908947.3 22797 22819 nCoV-2019_76_LEFT 2 +
MN908947.3 22798 22821 nCoV-2019_76_LEFT_alt3 2 +
MN908947.3 23192 23214 nCoV-2019_76_RIGHT 2 -
MN908947.3 23189 23212 nCoV-2019_76_RIGHT_alt0 2 -
MN908947.3 23122 23144 nCoV-2019_77_LEFT 1 +
MN908947.3 23500 23522 nCoV-2019_77_RIGHT 1 -
MN908947.3 23443 23466 nCoV-2019_78_LEFT 2 +
MN908947.3 23822 23847 nCoV-2019_78_RIGHT 2 -
MN908947.3 23789 23812 nCoV-2019_79_LEFT 1 +
MN908947.3 24145 24169 nCoV-2019_79_RIGHT 1 -
MN908947.3 24078 24100 nCoV-2019_80_LEFT 2 +
MN908947.3 24443 24467 nCoV-2019_80_RIGHT 2 -
MN908947.3 24391 24416 nCoV-2019_81_LEFT 1 +
MN908947.3 24765 24789 nCoV-2019_81_RIGHT 1 -
MN908947.3 24696 24721 nCoV-2019_82_LEFT 2 +
MN908947.3 25052 25076 nCoV-2019_82_RIGHT 2 -
MN908947.3 24978 25003 nCoV-2019_83_LEFT 1 +
MN908947.3 25347 25369 nCoV-2019_83_RIGHT 1 -
MN908947.3 25279 25301 nCoV-2019_84_LEFT 2 +
MN908947.3 25646 25673 nCoV-2019_84_RIGHT 2 -
MN908947.3 25601 25623 nCoV-2019_85_LEFT 1 +
MN908947.3 25969 25994 nCoV-2019_85_RIGHT 1 -
MN908947.3 25902 25924 nCoV-2019_86_LEFT 2 +
MN908947.3 26290 26315 nCoV-2019_86_RIGHT 2 -
MN908947.3 26197 26219 nCoV-2019_87_LEFT 1 +
MN908947.3 26566 26590 nCoV-2019_87_RIGHT 1 -
MN908947.3 26520 26542 nCoV-2019_88_LEFT 2 +
MN908947.3 26890 26913 nCoV-2019_88_RIGHT 2 -
MN908947.3 26835 26857 nCoV-2019_89_LEFT 1 +
MN908947.3 26838 26860 nCoV-2019_89_LEFT_alt2 1 +
MN908947.3 27202 27227 nCoV-2019_89_RIGHT 1 -
MN908947.3 27190 27215 nCoV-2019_89_RIGHT_alt4 1 -
MN908947.3 27141 27164 nCoV-2019_90_LEFT 2 +
MN908947.3 27511 27533 nCoV-2019_90_RIGHT 2 -
MN908947.3 27446 27471 nCoV-2019_91_LEFT 1 +
MN908947.3 27825 27854 nCoV-2019_91_RIGHT 1 -
MN908947.3 27784 27808 nCoV-2019_92_LEFT 2 +
MN908947.3 28145 28172 nCoV-2019_92_RIGHT 2 -
MN908947.3 28081 28104 nCoV-2019_93_LEFT 1 +
MN908947.3 28442 28464 nCoV-2019_93_RIGHT 1 -
MN908947.3 28394 28416 nCoV-2019_94_LEFT 2 +
MN908947.3 28756 28779 nCoV-2019_94_RIGHT 2 -
MN908947.3 28677 28699 nCoV-2019_95_LEFT 1 +
MN908947.3 29041 29063 nCoV-2019_95_RIGHT 1 -
MN908947.3 28985 29007 nCoV-2019_96_LEFT 2 +
MN908947.3 29356 29378 nCoV-2019_96_RIGHT 2 -
MN908947.3 29288 29316 nCoV-2019_97_LEFT 1 +
MN908947.3 29665 29693 nCoV-2019_97_RIGHT 1 -
MN908947.3 29486 29510 nCoV-2019_98_LEFT 2 +
MN908947.3 29836 29866 nCoV-2019_98_RIGHT 2 -
32 changes: 20 additions & 12 deletions workflow/rules/common.smk
View file
Original file line number Diff line number Diff line change
Expand Up @@ -355,9 +355,11 @@ def get_non_cov2_calls(from_caller="pangolin"):
pattern = (
"results/benchmarking/tables/strain-calls/non-cov2-{accession}.polished.strains.pangolin.csv"
if from_caller == "pangolin"
else "results/benchmarking/tables/strain-calls/non-cov2-{accession}.strains.kallisto.tsv"
if from_caller == "kallisto"
else []
else (
"results/benchmarking/tables/strain-calls/non-cov2-{accession}.strains.kallisto.tsv"
if from_caller == "kallisto"
else []
)
)

if not pattern:
Expand Down Expand Up @@ -1113,18 +1115,22 @@ def get_fallbacks_for_report(fallback_type):
if fallback_type == "pseudo":
path = "results/{{date}}/contigs/pseudoassembled/{sample}.fasta"
return [
path.format(sample=sample)
if has_pseudo_assembly(None, sample)
else "resources/genomes/main.fasta"
(
path.format(sample=sample)
if has_pseudo_assembly(None, sample)
else "resources/genomes/main.fasta"
)
for sample in get_samples_for_date(wildcards.date)
]

elif fallback_type == "consensus":
path = "results/{{date}}/contigs/masked/consensus/{sample}.fasta"
return [
path.format(sample=sample)
if has_consensus_assembly(None, sample)
else "resources/genomes/main.fasta"
(
path.format(sample=sample)
if has_consensus_assembly(None, sample)
else "resources/genomes/main.fasta"
)
for sample in samples
]

Expand Down Expand Up @@ -1390,9 +1396,11 @@ def get_kallisto_quant_extra(wildcards, input):
return get_if_testing("--single --fragment-length 250 --sd 47301")

return (
f"--single --fragment-length {get_first_line(input.fragment_length)} --sd {get_first_line(input.standard_deviation)}"
if is_single_end(wildcards)
else "",
(
f"--single --fragment-length {get_first_line(input.fragment_length)} --sd {get_first_line(input.standard_deviation)}"
if is_single_end(wildcards)
else ""
),
)


Expand Down
8 changes: 5 additions & 3 deletions workflow/rules/generate_output.smk
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Original file line number Diff line number Diff line change
Expand Up @@ -381,9 +381,11 @@ rule snakemake_reports_patient:
"results/{date}/qc/laboratory/multiqc.html",
"results/{date}/plots/coverage-reference-genome.png",
"results/{date}/plots/coverage-assembled-genome.png",
lambda wildcards: "results/{date}/plots/primer-clipping-intervals.svg"
if any_sample_is_amplicon(wildcards)
else [],
lambda wildcards: (
"results/{date}/plots/primer-clipping-intervals.svg"
if any_sample_is_amplicon(wildcards)
else []
),
# 4. Assembly
"results/{date}/filter-overview",
"results/{date}/pangolin-call-overview",
Expand Down
2 changes: 1 addition & 1 deletion workflow/rules/read_clipping.smk
View file
Original file line number Diff line number Diff line change
Expand Up @@ -22,7 +22,7 @@ rule samtools_sort:

rule bed_to_bedpe:
input:
check_bed_for_URL(config["preprocessing"]["amplicon-primers"]),
config["preprocessing"]["amplicon-primers"],
output:
"resources/primer.bedpe",
log:
Expand Down
8 changes: 5 additions & 3 deletions workflow/rules/ref.smk
View file
Original file line number Diff line number Diff line change
Expand Up @@ -20,9 +20,11 @@ rule get_genome:
"resources/genomes/{accession}.fasta",
params:
accession=(
lambda w: config["virus-reference-genome"]
if w.accession == "main"
else w.accession
lambda w: (
config["virus-reference-genome"]
if w.accession == "main"
else w.accession
)
),
log:
"logs/genomes/get-genome/{accession}.log",
Expand Down
8 changes: 5 additions & 3 deletions workflow/rules/variant_calling.smk
View file
Original file line number Diff line number Diff line change
Expand Up @@ -75,9 +75,11 @@ rule longshot:
"results/{date}/candidate-calls/ref~{reference}/{sample}.homopolymer-longshot.vcf"
),
params:
reference_name=lambda w: config["virus-reference-genome"]
if w.reference == "main"
else f"{w.reference}.1",
reference_name=lambda w: (
config["virus-reference-genome"]
if w.reference == "main"
else f"{w.reference}.1"
),
log:
"logs/{date}/longshot/ref~{reference}/{sample}.log",
conda:
Expand Down
18 changes: 17 additions & 1 deletion workflow/scripts/bed-to-bedpe.py
View file
Original file line number Diff line number Diff line change
Expand Up @@ -4,10 +4,26 @@
# except according to those terms.

import pandas as pd
import urllib.request


# Function to check bed for url
def check_bed_for_URL(bed_file):
if "https" in bed_file:
filename = bed_file.split("/")[-1]
filepath = "resources/{}".format(filename)
urllib.request.urlretrieve(bed_file, filepath)
return filepath
else:
return bed_file


# Check bed file for URL
bed_file = check_bed_for_URL(snakemake.input[0])

# Function to create a bedpe file from a bed file
bed_list = []
with open(snakemake.input[0]) as f:
with open(bed_file) as f:
line = f.readlines()
for name in line:
bed_list.append(name.split())
Expand Down

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