This is a brief run through of the ARTIC pipeline. It covers:
We are using a jupyter notebook for this example so that we can host it on Binder. If you want to run the commands for yourself on the command line, you will need to remove the leading !
which is before all the code in this notebook (which is telling jupyter to execute a system command).
To run this notebook, you can click on each cell and press Run. Be sure to wait for the cell to complete before moving on to the next one. It might take a minute or so for each cell to complete.
If you are using this notebook via Binder, or the GitHub repository, the pipeline is already installed. For reference though, you can install the pipeline from conda:
conda create -n artic-env
conda activate artic-env
conda install -c bioconda -c conda-forge artic
We are using version 1.2.1, check you are on the same:
!artic -v
artic 1.2.1
To begin, we need some data. This repository already has some data for you to use, which was generated from a SARS-COV-2 positive control sample at the University of Birmingham. If you want to obtain the data for yourself, you can run the following:
wget http://artic.s3.climb.ac.uk/BHAM-Run88-PTC.fastq
This test data is only the FASTQ reads from the positive control sample. We have already basecalled, demuxed and filtered them from the original FAST5 data for this sample.
Because we only have FASTQ data, this example will use the medaka workflow of the ARTIC pipeline. This is because the nanopolish version requires FAST5 data as well as FASTQ.
To run the medaka workflow ARTIC pipeline on this data, we need to know:
{pore}_{device}_{caller variant}_{caller version}
As well as the FASTQ reads, we will also need:
Although the ARTIC pipeline will download these for us, we can also get them for ourselves in order to familiarise ourselves with them:
!artic-tools get_scheme --schemeVersion 3 scov2
[21:41:48] [artic-tools::get_scheme] starting primer scheme downloader [21:41:48] [artic-tools::get_scheme] requested scheme: scov2 [21:41:48] [artic-tools::get_scheme] requested version: 3 [21:41:48] [artic-tools::get_scheme] fetching manifest file [21:41:48] [artic-tools::get_scheme] ARTIC manifest URL: https://raw.githubusercontent.com/artic-network/primer-schemes/master/schemes_manifest.json [21:41:50] [artic-tools::get_scheme] ARTIC repository DOI: 10.5281/zenodo.4004423 [21:41:50] [artic-tools::get_scheme] finding primer scheme [21:41:50] [artic-tools::get_scheme] found requested scheme: sars-cov-2 (using alias scov2) [21:41:50] [artic-tools::get_scheme] downloading primer scheme [21:41:51] [artic-tools::get_scheme] saving primers to: scov2.v3.primer.bed [21:41:51] [artic-tools::get_scheme] saving reference to: scov2.v3.reference.fasta [21:41:51] [artic-tools::get_scheme] comparing checksums [21:41:51] [artic-tools::get_scheme] sha256 for primers: 6e98d7d5d1c6edac8ef0bac70d698e0828ae42bafe8f3bda0a6257d00ce414b5 [21:41:51] [artic-tools::get_scheme] sha256 for reference: b09a4a3d6824dc4a9f3a17d480f3335f73cb1507897f6dad0de871e8f00d8637
This will have downloaded the primer scheme (scov2.v3.primer.bed
) and the reference sequence (scov2.v3.reference.fasta
). You can get some information on the primer scheme using artic-tools:
!artic-tools validate_scheme scov2.v3.primer.bed
[21:41:56] [artic-tools::validate_scheme] starting primer scheme validator [21:41:56] [artic-tools::validate_scheme] reading scheme [21:41:56] [artic-tools::validate_scheme] collecting scheme stats [21:41:56] [artic-tools::validate_scheme] primer scheme file: scov2.v3.primer.bed [21:41:56] [artic-tools::validate_scheme] reference sequence: MN908947.3 [21:41:56] [artic-tools::validate_scheme] number of pools: 2 [21:41:56] [artic-tools::validate_scheme] number of primers: 218 (includes 22 alts) [21:41:56] [artic-tools::validate_scheme] minimum primer size: 22 [21:41:56] [artic-tools::validate_scheme] maximum primer size: 57 [21:41:56] [artic-tools::validate_scheme] number of amplicons: 98 [21:41:56] [artic-tools::validate_scheme] mean amplicon size: 343 [21:41:56] [artic-tools::validate_scheme] maximum amplicon size: 375 [21:41:56] [artic-tools::validate_scheme] scheme ref. span: 30-29866 [21:41:56] [artic-tools::validate_scheme] scheme overlaps: 12.850247%
The primer scheme file is in a BED format, where the columns equate to the following:
column | name | type | description |
---|---|---|---|
1 | chrom | string | primer reference sequence |
2 | chromStart | int | starting position of the primer in the reference sequence |
3 | chomEnd | int | ending position of the primer in the reference sequence |
4 | name | string | primer name |
5 | primerPool | int | primer pool* |
6 | strand | string (+/-) | primer direction |
* column 5 in the BED spec is an int for score, whereas here we are using it to denote primerPool.
If you want to look at the primer scheme file, we can do that here with some Python:
with open("scov2.v3.primer.bed", 'r') as f:
print(f.read())
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 -
Now we have the primer scheme, reference sequence and our FASTQ data. We can run the pipeline!
!artic minion \
--normalise 100 \
--threads 2 \
--medaka \
--medaka-model r941_min_high_g351 \
--strict \
--read-file ../data/BHAM-Run88-PTC.fastq.gz \
scov2/V3 \
my_example
could not find primer scheme and reference sequence, attempting to download Running: artic-tools get_scheme scov2 --schemeVersion 3 [21:42:18] [artic-tools::get_scheme] starting primer scheme downloader [21:42:18] [artic-tools::get_scheme] requested scheme: scov2 [21:42:18] [artic-tools::get_scheme] requested version: 3 [21:42:18] [artic-tools::get_scheme] fetching manifest file [21:42:18] [artic-tools::get_scheme] ARTIC manifest URL: https://raw.githubusercontent.com/artic-network/primer-schemes/master/schemes_manifest.json [21:42:19] [artic-tools::get_scheme] ARTIC repository DOI: 10.5281/zenodo.4004423 [21:42:19] [artic-tools::get_scheme] finding primer scheme [21:42:19] [artic-tools::get_scheme] found requested scheme: sars-cov-2 (using alias scov2) [21:42:19] [artic-tools::get_scheme] downloading primer scheme [21:42:19] [artic-tools::get_scheme] saving primers to: scov2.v3.primer.bed [21:42:19] [artic-tools::get_scheme] saving reference to: scov2.v3.reference.fasta [21:42:19] [artic-tools::get_scheme] comparing checksums [21:42:19] [artic-tools::get_scheme] sha256 for primers: 6e98d7d5d1c6edac8ef0bac70d698e0828ae42bafe8f3bda0a6257d00ce414b5 [21:42:19] [artic-tools::get_scheme] sha256 for reference: b09a4a3d6824dc4a9f3a17d480f3335f73cb1507897f6dad0de871e8f00d8637 Running: artic-tools validate_scheme ./primer-schemes/scov2/V3/scov2.scheme.bed [21:42:19] [artic-tools::validate_scheme] starting primer scheme validator [21:42:19] [artic-tools::validate_scheme] reading scheme [21:42:19] [artic-tools::validate_scheme] collecting scheme stats [21:42:19] [artic-tools::validate_scheme] primer scheme file: ./primer-schemes/scov2/V3/scov2.scheme.bed [21:42:19] [artic-tools::validate_scheme] reference sequence: MN908947.3 [21:42:19] [artic-tools::validate_scheme] number of pools: 2 [21:42:19] [artic-tools::validate_scheme] number of primers: 218 (includes 22 alts) [21:42:19] [artic-tools::validate_scheme] minimum primer size: 22 [21:42:19] [artic-tools::validate_scheme] maximum primer size: 57 [21:42:19] [artic-tools::validate_scheme] number of amplicons: 98 [21:42:19] [artic-tools::validate_scheme] mean amplicon size: 343 [21:42:19] [artic-tools::validate_scheme] maximum amplicon size: 375 [21:42:19] [artic-tools::validate_scheme] scheme ref. span: 30-29866 [21:42:19] [artic-tools::validate_scheme] scheme overlaps: 12.850247% Running: minimap2 -a -x map-ont -t 2 ./primer-schemes/scov2/V3/scov2.reference.fasta ../data/BHAM-Run88-PTC.fastq.gz | samtools view -bS -F 4 - | samtools sort -o my_example.sorted.bam - [M::mm_idx_gen::0.002*2.10] collected minimizers [M::mm_idx_gen::0.003*2.02] sorted minimizers [M::main::0.004*2.01] loaded/built the index for 1 target sequence(s) [M::mm_mapopt_update::0.004*1.94] mid_occ = 3 [M::mm_idx_stat] kmer size: 15; skip: 10; is_hpc: 0; #seq: 1 [M::mm_idx_stat::0.004*1.88] distinct minimizers: 5587 (99.93% are singletons); average occurrences: 1.004; average spacing: 5.332 [M::worker_pipeline::11.622*1.58] mapped 124656 sequences [M::main] Version: 2.17-r941 [M::main] CMD: minimap2 -a -x map-ont -t 2 ./primer-schemes/scov2/V3/scov2.reference.fasta ../data/BHAM-Run88-PTC.fastq.gz [M::main] Real time: 11.624 sec; CPU: 18.317 sec; Peak RSS: 0.165 GB Running: samtools index my_example.sorted.bam Running: align_trim --normalise 100 ./primer-schemes/scov2/V3/scov2.scheme.bed --start --remove-incorrect-pairs --report my_example.alignreport.txt < my_example.sorted.bam 2> my_example.alignreport.er | samtools sort -T my_example - -o my_example.trimmed.rg.sorted.bam Running: align_trim --normalise 100 ./primer-schemes/scov2/V3/scov2.scheme.bed --remove-incorrect-pairs --report my_example.alignreport.txt < my_example.sorted.bam 2> my_example.alignreport.er | samtools sort -T my_example - -o my_example.primertrimmed.rg.sorted.bam Running: samtools index my_example.trimmed.rg.sorted.bam Running: samtools index my_example.primertrimmed.rg.sorted.bam Running: medaka consensus --model r941_min_high_g351 --threads 2 --chunk_len 800 --chunk_ovlp 400 --RG 1 my_example.trimmed.rg.sorted.bam my_example.1.hdf [21:43:15 - medaka] Reads will be filtered to only those with RG tag: 1 [21:43:15 - Predict] Processing region(s): MN908947.3:0-29903 [21:43:15 - Predict] Setting tensorflow threads to 2. [21:43:15 - Predict] Processing 1 long region(s) with batching. [21:43:15 - Predict] Using model: /Users/willrowe/opt/miniconda3/envs/artic-pipeline-example/lib/python3.6/site-packages/medaka/data/r941_min_high_g351_model.hdf5. [21:43:15 - ModelLoad] Building model with cudnn optimization: False [21:43:16 - DLoader] Initializing data loader [21:43:16 - Sampler] Initializing sampler for consensus of region MN908947.3:0-29903. [21:43:16 - PWorker] Running inference for 0.0M draft bases. [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 30-409. [21:43:16 - Feature] Processed MN908947.3:30.0-409.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 642-1027. [21:43:16 - Feature] Processed MN908947.3:642.0-1027.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 1242-1650. [21:43:16 - Feature] Processed MN908947.3:1242.0-1650.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 1868-2268. [21:43:16 - Feature] Processed MN908947.3:1868.0-2268.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 2504-2903. [21:43:16 - Feature] Processed MN908947.3:2504.0-2903.0 (median depth 177.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 3144-3530. [21:43:16 - Feature] Processed MN908947.3:3144.0-3530.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 3771-4163. [21:43:16 - Feature] Processed MN908947.3:3771.0-4163.0 (median depth 192.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 4294-4689. [21:43:16 - Feature] Processed MN908947.3:4294.0-4689.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 4939-5320. [21:43:16 - Feature] Processed MN908947.3:4939.0-5320.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 5563-5956. [21:43:16 - Feature] Processed MN908947.3:5563.0-5956.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 6167-6549. [21:43:16 - Feature] Processed MN908947.3:6167.0-6549.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 6718-7116. [21:43:16 - Feature] Processed MN908947.3:6718.0-7116.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 7305-7693. [21:43:16 - Feature] Processed MN908947.3:7305.0-7693.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 7943-8340. [21:43:16 - Feature] Processed MN908947.3:7943.0-8340.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 8595-8982. [21:43:16 - Feature] Processed MN908947.3:8595.0-8982.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 9204-9584. [21:43:16 - Feature] Processed MN908947.3:9204.0-9584.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 9784-10170. [21:43:16 - Feature] Processed MN908947.3:9784.0-10170.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 10362-10762. [21:43:16 - Feature] Processed MN908947.3:10362.0-10762.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 10999-11393. [21:43:16 - Feature] Processed MN908947.3:10999.0-11393.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 11555-11948. [21:43:16 - Feature] Processed MN908947.3:11555.0-11948.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 12110-12489. [21:43:16 - Feature] Processed MN908947.3:12110.0-12489.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 12710-13095. [21:43:16 - Feature] Processed MN908947.3:12710.0-13095.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 13307-13698. [21:43:16 - Feature] Processed MN908947.3:13307.0-13698.0 (median depth 197.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 13918-14298. [21:43:16 - Feature] Processed MN908947.3:13918.0-14298.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 14545-14925. [21:43:16 - Feature] Processed MN908947.3:14545.0-14925.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 15171-15559. [21:43:16 - Feature] Processed MN908947.3:15171.0-15559.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 15827-16208. [21:43:16 - Feature] Processed MN908947.3:15827.0-16208.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 16416-16832. [21:43:16 - Feature] Processed MN908947.3:16416.0-16832.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 17065-17451. [21:43:16 - Feature] Processed MN908947.3:17065.0-17451.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 17674-18061. [21:43:16 - Feature] Processed MN908947.3:17674.0-18061.0 (median depth 162.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 18253-18671. [21:43:16 - Feature] Processed MN908947.3:18253.0-18671.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 18896-19296. [21:43:16 - Feature] Processed MN908947.3:18896.0-19296.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 19548-19938. [21:43:16 - Feature] Processed MN908947.3:19548.0-19938.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 20172-20571. [21:43:16 - Feature] Processed MN908947.3:20172.0-20571.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 20786-21168. [21:43:16 - Feature] Processed MN908947.3:20786.0-21168.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 21357-21742. [21:43:16 - Feature] Processed MN908947.3:21357.0-21742.0 (median depth 119.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 21961-22345. [21:43:16 - Feature] Processed MN908947.3:21961.0-22345.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 22516-22902. [21:43:16 - Feature] Processed MN908947.3:22516.0-22902.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 23122-23521. [21:43:16 - Feature] Processed MN908947.3:23122.0-23521.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 23789-24168. [21:43:16 - Feature] Processed MN908947.3:23789.0-24168.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 24391-24788. [21:43:16 - Feature] Processed MN908947.3:24391.0-24788.2 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 24978-25368. [21:43:16 - Feature] Processed MN908947.3:24978.0-25368.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 25601-25993. [21:43:16 - Feature] Processed MN908947.3:25601.0-25993.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 26197-26589. [21:43:16 - Feature] Processed MN908947.3:26197.0-26589.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 26835-27226. [21:43:16 - Feature] Processed MN908947.3:26835.0-27226.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 27446-27853. [21:43:16 - Feature] Processed MN908947.3:27446.0-27853.0 (median depth 91.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 28081-28463. [21:43:16 - Feature] Processed MN908947.3:28081.0-28463.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 28677-29062. [21:43:16 - Feature] Processed MN908947.3:28677.0-29062.0 (median depth 200.0) [21:43:16 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 29288-29692. [21:43:16 - Feature] Processed MN908947.3:29288.0-29692.0 (median depth 200.0) [21:43:16 - Sampler] Took 0.15s to make features. [21:43:16 - Sampler] Region MN908947.3:2504.0-2903.0 (794 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:3771.0-4163.0 (771 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:4939.0-5320.0 (770 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:6167.0-6549.0 (771 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:8595.0-8982.0 (778 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:10999.0-11393.0 (757 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:12110.0-12489.0 (748 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:12710.0-13095.0 (793 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:13918.0-14298.0 (777 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:17674.0-18061.0 (750 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:20786.0-21168.0 (797 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:21357.0-21742.0 (714 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:22516.0-22902.0 (787 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:23789.0-24168.0 (773 positions) is smaller than inference chunk length 800, quarantining. [21:43:16 - Sampler] Region MN908947.3:27446.0-27853.0 (656 positions) is smaller than inference chunk length 800, quarantining. [21:43:17 - PWorker] All done, 15 remainder regions. [21:43:17 - Predict] Processing 15 short region(s). [21:43:17 - ModelLoad] Building model with cudnn optimization: False [21:43:18 - DLoader] Initializing data loader [21:43:18 - PWorker] Running inference for 0.0M draft bases. [21:43:18 - Sampler] Initializing sampler for consensus of region MN908947.3:2504-2904. [21:43:18 - Feature] Processed MN908947.3:2504.0-2903.0 (median depth 177.0) [21:43:18 - Sampler] Took 0.01s to make features. [21:43:19 - PWorker] All done, 0 remainder regions. [21:43:19 - DLoader] Initializing data loader [21:43:19 - Sampler] Initializing sampler for consensus of region MN908947.3:3771-4164. [21:43:19 - PWorker] Running inference for 0.0M draft bases. [21:43:19 - Feature] Processed MN908947.3:3771.0-4163.0 (median depth 192.0) [21:43:19 - Sampler] Took 0.02s to make features. [21:43:20 - PWorker] All done, 0 remainder regions. [21:43:20 - DLoader] Initializing data loader [21:43:20 - Sampler] Initializing sampler for consensus of region MN908947.3:4939-5321. [21:43:20 - PWorker] Running inference for 0.0M draft bases. [21:43:20 - Feature] Processed MN908947.3:4939.0-5320.0 (median depth 200.0) [21:43:20 - Sampler] Took 0.02s to make features. [21:43:20 - PWorker] All done, 0 remainder regions. [21:43:20 - DLoader] Initializing data loader [21:43:20 - Sampler] Initializing sampler for consensus of region MN908947.3:6167-6550. [21:43:20 - PWorker] Running inference for 0.0M draft bases. [21:43:21 - Feature] Processed MN908947.3:6167.0-6549.0 (median depth 200.0) [21:43:21 - Sampler] Took 0.01s to make features. [21:43:21 - PWorker] All done, 0 remainder regions. [21:43:21 - DLoader] Initializing data loader [21:43:21 - Sampler] Initializing sampler for consensus of region MN908947.3:8595-8983. [21:43:21 - PWorker] Running inference for 0.0M draft bases. [21:43:21 - Feature] Processed MN908947.3:8595.0-8982.0 (median depth 200.0) [21:43:21 - Sampler] Took 0.02s to make features. [21:43:21 - PWorker] All done, 0 remainder regions. [21:43:21 - DLoader] Initializing data loader [21:43:21 - Sampler] Initializing sampler for consensus of region MN908947.3:10999-11394. [21:43:21 - PWorker] Running inference for 0.0M draft bases. [21:43:21 - Feature] Processed MN908947.3:10999.0-11393.0 (median depth 200.0) [21:43:21 - Sampler] Took 0.02s to make features. [21:43:21 - PWorker] All done, 0 remainder regions. [21:43:21 - DLoader] Initializing data loader [21:43:21 - PWorker] Running inference for 0.0M draft bases. [21:43:21 - Sampler] Initializing sampler for consensus of region MN908947.3:12110-12490. [21:43:21 - Feature] Processed MN908947.3:12110.0-12489.0 (median depth 200.0) [21:43:21 - Sampler] Took 0.02s to make features. [21:43:22 - PWorker] All done, 0 remainder regions. [21:43:22 - DLoader] Initializing data loader [21:43:22 - PWorker] Running inference for 0.0M draft bases. [21:43:22 - Sampler] Initializing sampler for consensus of region MN908947.3:12710-13096. [21:43:22 - Feature] Processed MN908947.3:12710.0-13095.0 (median depth 200.0) [21:43:22 - Sampler] Took 0.02s to make features. [21:43:22 - PWorker] All done, 0 remainder regions. [21:43:22 - DLoader] Initializing data loader [21:43:22 - PWorker] Running inference for 0.0M draft bases. [21:43:22 - Sampler] Initializing sampler for consensus of region MN908947.3:13918-14299. [21:43:22 - Feature] Processed MN908947.3:13918.0-14298.0 (median depth 200.0) [21:43:22 - Sampler] Took 0.03s to make features. [21:43:23 - PWorker] All done, 0 remainder regions. [21:43:23 - DLoader] Initializing data loader [21:43:23 - Sampler] Initializing sampler for consensus of region MN908947.3:17674-18062. [21:43:23 - PWorker] Running inference for 0.0M draft bases. [21:43:23 - Feature] Processed MN908947.3:17674.0-18061.0 (median depth 162.0) [21:43:23 - Sampler] Took 0.01s to make features. [21:43:23 - PWorker] All done, 0 remainder regions. [21:43:23 - DLoader] Initializing data loader [21:43:23 - Sampler] Initializing sampler for consensus of region MN908947.3:20786-21169. [21:43:23 - PWorker] Running inference for 0.0M draft bases. [21:43:23 - Feature] Processed MN908947.3:20786.0-21168.0 (median depth 200.0) [21:43:23 - Sampler] Took 0.03s to make features. [21:43:24 - PWorker] All done, 0 remainder regions. [21:43:24 - DLoader] Initializing data loader [21:43:24 - PWorker] Running inference for 0.0M draft bases. [21:43:24 - Sampler] Initializing sampler for consensus of region MN908947.3:21357-21743. [21:43:24 - Feature] Processed MN908947.3:21357.0-21742.0 (median depth 119.0) [21:43:24 - Sampler] Took 0.03s to make features. [21:43:25 - PWorker] All done, 0 remainder regions. [21:43:25 - DLoader] Initializing data loader [21:43:25 - Sampler] Initializing sampler for consensus of region MN908947.3:22516-22903. [21:43:25 - PWorker] Running inference for 0.0M draft bases. [21:43:25 - Feature] Processed MN908947.3:22516.0-22902.0 (median depth 200.0) [21:43:25 - Sampler] Took 0.03s to make features. [21:43:25 - PWorker] All done, 0 remainder regions. [21:43:25 - DLoader] Initializing data loader [21:43:25 - PWorker] Running inference for 0.0M draft bases. [21:43:25 - Sampler] Initializing sampler for consensus of region MN908947.3:23789-24169. [21:43:26 - Feature] Processed MN908947.3:23789.0-24168.0 (median depth 200.0) [21:43:26 - Sampler] Took 0.03s to make features. [21:43:26 - PWorker] All done, 0 remainder regions. [21:43:26 - DLoader] Initializing data loader [21:43:26 - Sampler] Initializing sampler for consensus of region MN908947.3:27446-27854. [21:43:26 - PWorker] Running inference for 0.0M draft bases. [21:43:26 - Feature] Processed MN908947.3:27446.0-27853.0 (median depth 91.0) [21:43:26 - Sampler] Took 0.04s to make features. [21:43:27 - PWorker] All done, 0 remainder regions. [21:43:27 - Predict] Finished processing all regions. Running: medaka variant ./primer-schemes/scov2/V3/scov2.reference.fasta my_example.1.hdf my_example.1.vcf [21:43:32 - DataIndex] Loaded 1/1 (100.00%) sample files. [21:43:32 - Variants] Processing MN908947.3:0-. [21:43:32 - TrimOverlap] MN908947.3:56.1-409.0 and MN908947.3:642.0-984.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:683.2-1027.0 and MN908947.3:1242.0-1613.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:1274.0-1650.0 and MN908947.3:1868.0-2236.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:1894.0-2268.0 and MN908947.3:2504.0-2903.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:2504.0-2903.0 and MN908947.3:3144.0-3514.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:3155.0-3530.0 and MN908947.3:3771.0-4163.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:3771.0-4163.0 and MN908947.3:4294.0-4678.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:4303.5-4689.0 and MN908947.3:4939.0-5320.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:4939.0-5320.0 and MN908947.3:5563.0-5920.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:5599.1-5956.0 and MN908947.3:6167.0-6549.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:6167.0-6549.0 and MN908947.3:6718.0-7076.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:6757.1-7116.0 and MN908947.3:7305.0-7669.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:32 - TrimOverlap] MN908947.3:7328.0-7693.0 and MN908947.3:7943.0-8311.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:7965.2-8340.0 and MN908947.3:8595.0-8982.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:8595.0-8982.0 and MN908947.3:9204.0-9558.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:9229.1-9584.0 and MN908947.3:9784.0-10152.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:9801.0-10170.0 and MN908947.3:10362.0-10750.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:10374.0-10762.0 and MN908947.3:10999.0-11393.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:10999.0-11393.0 and MN908947.3:11555.0-11925.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:11576.0-11948.0 and MN908947.3:12110.0-12489.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:12110.0-12489.0 and MN908947.3:12710.0-13095.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:12710.0-13095.0 and MN908947.3:13307.0-13670.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:13336.2-13698.0 and MN908947.3:13918.0-14298.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:13918.0-14298.0 and MN908947.3:14545.0-14918.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:14551.1-14925.0 and MN908947.3:15171.0-15553.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:15175.1-15559.0 and MN908947.3:15827.0-16202.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:15833.3-16208.0 and MN908947.3:16416.0-16802.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:16452.0-16832.0 and MN908947.3:17065.0-17401.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:17115.1-17451.0 and MN908947.3:17674.0-18061.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:17674.0-18061.0 and MN908947.3:18253.0-18604.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:18316.1-18671.0 and MN908947.3:18896.0-19255.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:18934.1-19296.0 and MN908947.3:19548.0-19920.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:19559.0-19938.0 and MN908947.3:20172.0-20554.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:20189.0-20571.0 and MN908947.3:20786.0-21168.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:20786.0-21168.0 and MN908947.3:21357.0-21742.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:21357.0-21742.0 and MN908947.3:21961.0-22340.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:21968.0-22345.0 and MN908947.3:22516.0-22902.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:22516.0-22902.0 and MN908947.3:23122.0-23497.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:23154.1-23521.0 and MN908947.3:23789.0-24168.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:23789.0-24168.0 and MN908947.3:24391.0-24755.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:24419.1-24788.2 and MN908947.3:24978.0-25356.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:24986.3-25368.0 and MN908947.3:25601.0-25948.9 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:25632.1-25993.0 and MN908947.3:26197.0-26516.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:26274.3-26589.0 and MN908947.3:26835.0-27188.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:26863.1-27226.0 and MN908947.3:27446.0-27853.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:27446.0-27853.0 and MN908947.3:28081.0-28445.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:28102.0-28463.0 and MN908947.3:28677.0-29035.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:33 - TrimOverlap] MN908947.3:28705.2-29062.0 and MN908947.3:29288.0-29642.1 cannot be concatenated as there is no overlap and they do not abut. Running: medaka consensus --model r941_min_high_g351 --threads 2 --chunk_len 800 --chunk_ovlp 400 --RG 2 my_example.trimmed.rg.sorted.bam my_example.2.hdf [21:43:37 - medaka] Reads will be filtered to only those with RG tag: 2 [21:43:37 - Predict] Processing region(s): MN908947.3:0-29903 [21:43:37 - Predict] Setting tensorflow threads to 2. [21:43:37 - Predict] Processing 1 long region(s) with batching. [21:43:37 - Predict] Using model: /Users/willrowe/opt/miniconda3/envs/artic-pipeline-example/lib/python3.6/site-packages/medaka/data/r941_min_high_g351_model.hdf5. [21:43:37 - ModelLoad] Building model with cudnn optimization: False [21:43:39 - DLoader] Initializing data loader [21:43:39 - Sampler] Initializing sampler for consensus of region MN908947.3:0-29903. [21:43:39 - PWorker] Running inference for 0.0M draft bases. [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 320-725. [21:43:39 - Feature] Processed MN908947.3:320.0-725.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 943-1336. [21:43:39 - Feature] Processed MN908947.3:943.0-1336.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 1573-1963. [21:43:39 - Feature] Processed MN908947.3:1573.0-1963.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 2181-2591. [21:43:39 - Feature] Processed MN908947.3:2181.0-2591.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 2826-3209. [21:43:39 - Feature] Processed MN908947.3:2826.0-3209.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 3460-3852. [21:43:39 - Feature] Processed MN908947.3:3460.0-3852.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 4044-4449. [21:43:39 - Feature] Processed MN908947.3:4044.0-4449.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 4636-5016. [21:43:39 - Feature] Processed MN908947.3:4636.0-5016.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 5230-5643. [21:43:39 - Feature] Processed MN908947.3:5230.0-5643.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 5867-6271. [21:43:39 - Feature] Processed MN908947.3:5867.0-6271.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 6466-6872. [21:43:39 - Feature] Processed MN908947.3:6466.0-6872.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 7035-7414. [21:43:39 - Feature] Processed MN908947.3:7035.0-7414.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 7626-8018. [21:43:39 - Feature] Processed MN908947.3:7626.0-8018.0 (median depth 117.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 8249-8660. [21:43:39 - Feature] Processed MN908947.3:8249.0-8660.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 8888-9270. [21:43:39 - Feature] Processed MN908947.3:8888.0-9270.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 9477-9857. [21:43:39 - Feature] Processed MN908947.3:9477.0-9857.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 10076-10458. [21:43:39 - Feature] Processed MN908947.3:10076.0-10458.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 10666-11073. [21:43:39 - Feature] Processed MN908947.3:10666.0-11073.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 11306-11692. [21:43:39 - Feature] Processed MN908947.3:11306.0-11692.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 11863-12255. [21:43:39 - Feature] Processed MN908947.3:11863.0-12255.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 12417-12801. [21:43:39 - Feature] Processed MN908947.3:12417.0-12801.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 13005-13399. [21:43:39 - Feature] Processed MN908947.3:13005.0-13399.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 13599-13983. [21:43:39 - Feature] Processed MN908947.3:13599.0-13983.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 14207-14600. [21:43:39 - Feature] Processed MN908947.3:14207.0-14600.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 14865-15245. [21:43:39 - Feature] Processed MN908947.3:14865.0-15245.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 15481-15885. [21:43:39 - Feature] Processed MN908947.3:15481.0-15885.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 16118-16509. [21:43:39 - Feature] Processed MN908947.3:16118.0-16509.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 16748-17151. [21:43:39 - Feature] Processed MN908947.3:16748.0-17151.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 17381-17760. [21:43:39 - Feature] Processed MN908947.3:17381.0-17760.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 17966-18347. [21:43:39 - Feature] Processed MN908947.3:17966.0-18347.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 18596-18978. [21:43:39 - Feature] Processed MN908947.3:18596.0-18978.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 19204-19615. [21:43:39 - Feature] Processed MN908947.3:19204.0-19615.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 19844-20254. [21:43:39 - Feature] Processed MN908947.3:19844.0-20254.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 20472-20889. [21:43:39 - Feature] Processed MN908947.3:20472.0-20889.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 21075-21454. [21:43:39 - Feature] Processed MN908947.3:21075.0-21454.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 21658-22037. [21:43:39 - Feature] Processed MN908947.3:21658.0-22037.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 22262-22649. [21:43:39 - Feature] Processed MN908947.3:22262.0-22649.0 (median depth 34.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 22797-23213. [21:43:39 - Feature] Processed MN908947.3:22797.0-23213.0 (median depth 121.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 23443-23846. [21:43:39 - Feature] Processed MN908947.3:23443.0-23846.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 24078-24466. [21:43:39 - Feature] Processed MN908947.3:24078.0-24466.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 24696-25075. [21:43:39 - Feature] Processed MN908947.3:24696.0-25075.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 25279-25672. [21:43:39 - Feature] Processed MN908947.3:25279.0-25672.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 25902-26314. [21:43:39 - Feature] Processed MN908947.3:25902.0-26314.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 26520-26912. [21:43:39 - Feature] Processed MN908947.3:26520.0-26912.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 27141-27532. [21:43:39 - Feature] Processed MN908947.3:27141.0-27532.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 27784-28171. [21:43:39 - Feature] Processed MN908947.3:27784.0-28171.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 28394-28778. [21:43:39 - Feature] Processed MN908947.3:28394.0-28778.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 28985-29377. [21:43:39 - Feature] Processed MN908947.3:28985.0-29377.0 (median depth 200.0) [21:43:39 - Feature] Pileup counts do not span requested region, requested MN908947.3:0-29903, received 29486-29865. [21:43:39 - Feature] Processed MN908947.3:29486.0-29865.0 (median depth 200.0) [21:43:39 - Sampler] Took 0.14s to make features. [21:43:39 - Sampler] Region MN908947.3:4044.0-4449.0 (794 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:4636.0-5016.0 (784 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:7035.0-7414.0 (759 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:7626.0-8018.0 (687 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:12417.0-12801.0 (778 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:14865.0-15245.0 (779 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:17381.0-17760.0 (793 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:21075.0-21454.0 (799 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:22262.0-22649.0 (488 positions) is smaller than inference chunk length 800, quarantining. [21:43:39 - Sampler] Region MN908947.3:22797.0-23213.0 (687 positions) is smaller than inference chunk length 800, quarantining. [21:43:42 - PWorker] All done, 10 remainder regions. [21:43:42 - Predict] Processing 10 short region(s). [21:43:42 - ModelLoad] Building model with cudnn optimization: False [21:43:44 - DLoader] Initializing data loader [21:43:44 - Sampler] Initializing sampler for consensus of region MN908947.3:4044-4450. [21:43:44 - PWorker] Running inference for 0.0M draft bases. [21:43:44 - Feature] Processed MN908947.3:4044.0-4449.0 (median depth 200.0) [21:43:44 - Sampler] Took 0.01s to make features. [21:43:44 - PWorker] All done, 0 remainder regions. [21:43:44 - DLoader] Initializing data loader [21:43:44 - PWorker] Running inference for 0.0M draft bases. [21:43:44 - Sampler] Initializing sampler for consensus of region MN908947.3:4636-5017. [21:43:44 - Feature] Processed MN908947.3:4636.0-5016.0 (median depth 200.0) [21:43:44 - Sampler] Took 0.02s to make features. [21:43:45 - PWorker] All done, 0 remainder regions. [21:43:45 - DLoader] Initializing data loader [21:43:45 - PWorker] Running inference for 0.0M draft bases. [21:43:45 - Sampler] Initializing sampler for consensus of region MN908947.3:7035-7415. [21:43:45 - Feature] Processed MN908947.3:7035.0-7414.0 (median depth 200.0) [21:43:45 - Sampler] Took 0.02s to make features. [21:43:45 - PWorker] All done, 0 remainder regions. [21:43:45 - DLoader] Initializing data loader [21:43:45 - PWorker] Running inference for 0.0M draft bases. [21:43:45 - Sampler] Initializing sampler for consensus of region MN908947.3:7626-8019. [21:43:45 - Feature] Processed MN908947.3:7626.0-8018.0 (median depth 117.0) [21:43:45 - Sampler] Took 0.02s to make features. [21:43:45 - PWorker] All done, 0 remainder regions. [21:43:45 - DLoader] Initializing data loader [21:43:45 - Sampler] Initializing sampler for consensus of region MN908947.3:12417-12802. [21:43:45 - PWorker] Running inference for 0.0M draft bases. [21:43:45 - Feature] Processed MN908947.3:12417.0-12801.0 (median depth 200.0) [21:43:45 - Sampler] Took 0.02s to make features. [21:43:46 - PWorker] All done, 0 remainder regions. [21:43:46 - DLoader] Initializing data loader [21:43:46 - Sampler] Initializing sampler for consensus of region MN908947.3:14865-15246. [21:43:46 - PWorker] Running inference for 0.0M draft bases. [21:43:46 - Feature] Processed MN908947.3:14865.0-15245.0 (median depth 200.0) [21:43:46 - Sampler] Took 0.03s to make features. [21:43:46 - PWorker] All done, 0 remainder regions. [21:43:46 - DLoader] Initializing data loader [21:43:46 - Sampler] Initializing sampler for consensus of region MN908947.3:17381-17761. [21:43:46 - PWorker] Running inference for 0.0M draft bases. [21:43:46 - Feature] Processed MN908947.3:17381.0-17760.0 (median depth 200.0) [21:43:46 - Sampler] Took 0.01s to make features. [21:43:46 - PWorker] All done, 0 remainder regions. [21:43:46 - DLoader] Initializing data loader [21:43:46 - Sampler] Initializing sampler for consensus of region MN908947.3:21075-21455. [21:43:46 - PWorker] Running inference for 0.0M draft bases. [21:43:46 - Feature] Processed MN908947.3:21075.0-21454.0 (median depth 200.0) [21:43:46 - Sampler] Took 0.01s to make features. [21:43:46 - PWorker] All done, 0 remainder regions. [21:43:46 - DLoader] Initializing data loader [21:43:46 - PWorker] Running inference for 0.0M draft bases. [21:43:46 - Sampler] Initializing sampler for consensus of region MN908947.3:22262-22650. [21:43:47 - Feature] Processed MN908947.3:22262.0-22649.0 (median depth 34.0) [21:43:47 - Sampler] Took 0.01s to make features. [21:43:47 - PWorker] All done, 0 remainder regions. [21:43:47 - DLoader] Initializing data loader [21:43:47 - PWorker] Running inference for 0.0M draft bases. [21:43:47 - Sampler] Initializing sampler for consensus of region MN908947.3:22797-23214. [21:43:47 - Feature] Processed MN908947.3:22797.0-23213.0 (median depth 121.0) [21:43:47 - Sampler] Took 0.02s to make features. [21:43:48 - PWorker] All done, 0 remainder regions. [21:43:48 - Predict] Finished processing all regions. Running: medaka variant ./primer-schemes/scov2/V3/scov2.reference.fasta my_example.2.hdf my_example.2.vcf [21:43:53 - DataIndex] Loaded 1/1 (100.00%) sample files. [21:43:53 - Variants] Processing MN908947.3:0-. [21:43:53 - TrimOverlap] MN908947.3:366.2-725.0 and MN908947.3:943.0-1291.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:989.1-1336.0 and MN908947.3:1573.0-1958.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:1578.0-1963.0 and MN908947.3:2181.0-2546.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:2219.1-2591.0 and MN908947.3:2826.0-3193.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:2838.0-3209.0 and MN908947.3:3460.0-3810.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:3498.2-3852.0 and MN908947.3:4044.0-4449.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:4044.0-4449.0 and MN908947.3:4636.0-5016.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:4636.0-5016.0 and MN908947.3:5230.0-5630.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:5250.0-5643.0 and MN908947.3:5867.0-6256.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:5877.4-6271.0 and MN908947.3:6466.0-6858.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:6479.1-6872.0 and MN908947.3:7035.0-7414.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:7035.0-7414.0 and MN908947.3:7626.0-8018.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:7626.0-8018.0 and MN908947.3:8249.0-8622.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:8291.0-8660.0 and MN908947.3:8888.0-9263.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:8893.3-9270.0 and MN908947.3:9477.0-9794.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:9540.1-9857.0 and MN908947.3:10076.0-10449.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:10083.0-10458.0 and MN908947.3:10666.0-11056.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:10679.1-11073.0 and MN908947.3:11306.0-11672.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:11325.2-11692.0 and MN908947.3:11863.0-12246.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:11872.3-12255.0 and MN908947.3:12417.0-12801.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:12417.0-12801.0 and MN908947.3:13005.0-13368.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:13027.1-13399.0 and MN908947.3:13599.0-13975.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:13604.2-13983.0 and MN908947.3:14207.0-14580.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:14230.0-14600.0 and MN908947.3:14865.0-15245.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:14865.0-15245.0 and MN908947.3:15481.0-15862.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:15501.1-15885.0 and MN908947.3:16118.0-16483.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:16145.3-16509.0 and MN908947.3:16748.0-17112.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:16790.0-17151.0 and MN908947.3:17381.0-17760.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:17381.0-17760.0 and MN908947.3:17966.0-18321.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:17985.0-18347.0 and MN908947.3:18596.0-18972.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:18599.2-18978.0 and MN908947.3:19204.0-19540.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:19272.1-19615.0 and MN908947.3:19844.0-20219.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:19868.5-20254.0 and MN908947.3:20472.0-20865.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:20497.1-20889.0 and MN908947.3:21075.0-21454.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:21075.0-21454.0 and MN908947.3:21658.0-22024.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:21669.1-22037.0 and MN908947.3:22262.0-22649.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:22262.0-22649.0 and MN908947.3:22797.0-23213.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:22797.0-23213.0 and MN908947.3:23443.0-23765.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:23510.2-23846.0 and MN908947.3:24078.0-24445.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:53 - TrimOverlap] MN908947.3:24092.1-24466.0 and MN908947.3:24696.0-25044.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:24720.1-25075.0 and MN908947.3:25279.0-25642.0 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:25302.0-25672.0 and MN908947.3:25902.0-26253.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:25954.2-26314.0 and MN908947.3:26520.0-26895.3 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:26539.2-26912.0 and MN908947.3:27141.0-27522.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:27152.1-27532.0 and MN908947.3:27784.0-28160.1 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:27795.1-28171.0 and MN908947.3:28394.0-28756.9 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:28418.2-28778.0 and MN908947.3:28985.0-29352.2 cannot be concatenated as there is no overlap and they do not abut. [21:43:54 - TrimOverlap] MN908947.3:29015.1-29377.0 and MN908947.3:29486.0-29859.1 cannot be concatenated as there is no overlap and they do not abut. Running: artic_vcf_merge my_example ./primer-schemes/scov2/V3/scov2.scheme.bed 2> my_example.primersitereport.txt 1:my_example.1.vcf 2:my_example.2.vcf Running: bgzip -f my_example.merged.vcf Running: tabix -p vcf my_example.merged.vcf.gz Running: artic-tools check_vcf --dropPrimerVars --dropOverlapFails --vcfOut my_example.merged.filtered.vcf my_example.merged.vcf.gz ./primer-schemes/scov2/V3/scov2.scheme.bed 2> my_example.vcfreport.txt Running: mv my_example.merged.filtered.vcf my_example.merged.vcf Running: bgzip -f my_example.merged.vcf Running: tabix -f -p vcf my_example.merged.vcf.gz Running: longshot -P 0 -F -A --no_haps --bam my_example.primertrimmed.rg.sorted.bam --ref ./primer-schemes/scov2/V3/scov2.reference.fasta --out my_example.merged.vcf --potential_variants my_example.merged.vcf.gz 2021-01-14 21:43:56 Automatically determining max read coverage. 2021-01-14 21:43:56 Estimating mean read coverage... 2021-01-14 21:43:56 Total reference positions: 29903 2021-01-14 21:43:56 Total bases in bam: 6503829 2021-01-14 21:43:56 Mean read coverage: 217.50 2021-01-14 21:43:56 Min read coverage set to 6. 2021-01-14 21:43:56 Max read coverage set to 291. 2021-01-14 21:43:56 Estimating alignment parameters... 2021-01-14 21:43:56 Done estimating alignment parameters. Transition Probabilities: match -> match: 0.983 match -> insertion: 0.007 match -> deletion: 0.010 deletion -> match: 0.647 deletion -> deletion: 0.353 insertion -> match: 0.673 insertion -> insertion: 0.327 Emission Probabilities: match (equal): 0.987 match (not equal): 0.004 insertion: 1.000 deletion: 1.000 GENOTYPE PRIORS: REF G1/G2 PROB C D/I 0.0000000000015624993116907664 G A/A 0.00016666692910805806 G D/I 0.0000000000015624993116907664 T T/T 0.9984401606149776 A C/D 0.0000000004166671407128834 A A/T 0.0003333338549006453 C A/A 0.00016666692910805806 G T/T 0.00016666692910805806 C C/T 0.0003333338549006453 T A/T 0.0003333338549006453 G G/I 0.000001250002950856874 T D/I 0.0000000000015624993116907664 C D/D 0.000006249988919003825 T G/D 0.0000000004166671407128834 C A/T 0.00000011111137065235802 C G/D 0.0000000004166671407128834 G T/I 0.0000000004166671407128834 C C/C 0.9984401606149776 A G/G 0.00016666692910805806 A A/I 0.000001250002950856874 A A/C 0.0003333338549006453 T T/I 0.000001250002950856874 G G/D 0.000001250002950856874 T C/G 0.00000011111137065235802 G C/G 0.0003333338549006453 G A/I 0.0000000004166671407128834 G C/I 0.0000000004166671407128834 C T/D 0.0000000004166671407128834 G A/C 0.00000011111137065235802 A I/I 0.000006249988919003825 A G/T 0.00000011111137065235802 C G/I 0.0000000004166671407128834 T A/C 0.00000011111137065235802 T A/I 0.0000000004166671407128834 C A/G 0.00000011111137065235802 T T/D 0.000001250002950856874 A C/T 0.00000011111137065235802 A A/D 0.000001250002950856874 A T/T 0.00016666692910805806 A D/I 0.0000000000015624993116907664 G A/T 0.00000011111137065235802 G G/G 0.9984401606149776 G C/T 0.00000011111137065235802 G I/I 0.000006249988919003825 T A/D 0.0000000004166671407128834 T C/D 0.0000000004166671407128834 T I/I 0.000006249988919003825 A C/G 0.00000011111137065235802 C C/G 0.0003333338549006453 C A/D 0.0000000004166671407128834 C G/T 0.00000011111137065235802 T D/D 0.000006249988919003825 A A/G 0.0003333338549006453 T C/I 0.0000000004166671407128834 A D/D 0.000006249988919003825 T A/A 0.00016666692910805806 T G/I 0.0000000004166671407128834 G G/T 0.0003333338549006453 C G/G 0.00016666692910805806 G T/D 0.0000000004166671407128834 C C/D 0.000001250002950856874 A G/D 0.0000000004166671407128834 C A/C 0.0003333338549006453 C A/I 0.0000000004166671407128834 T G/T 0.0003333338549006453 A T/D 0.0000000004166671407128834 G C/D 0.0000000004166671407128834 G A/D 0.0000000004166671407128834 C T/I 0.0000000004166671407128834 C C/I 0.000001250002950856874 T C/T 0.0003333338549006453 A C/I 0.0000000004166671407128834 A C/C 0.00016666692910805806 C I/I 0.000006249988919003825 G D/D 0.000006249988919003825 A A/A 0.9984401606149776 A G/I 0.0000000004166671407128834 C T/T 0.00016666692910805806 T C/C 0.00016666692910805806 G C/C 0.00016666692910805806 T A/G 0.00000011111137065235802 T G/G 0.00016666692910805806 A T/I 0.0000000004166671407128834 G A/G 0.0003333338549006453 2021-01-14 21:43:56 Reading potential variants from input VCF... 2021-01-14 21:43:56 13 potential variants identified. 2021-01-14 21:43:56 Generating haplotype fragments from reads... 2021-01-14 21:43:56 10% of variants processed... 2021-01-14 21:43:56 20% of variants processed... 2021-01-14 21:43:56 30% of variants processed... 2021-01-14 21:43:56 40% of variants processed... 2021-01-14 21:43:56 50% of variants processed... 2021-01-14 21:43:57 60% of variants processed... 2021-01-14 21:43:57 70% of variants processed... 2021-01-14 21:43:57 80% of variants processed... 2021-01-14 21:43:57 100% of variants processed. 2021-01-14 21:43:57 Calling initial genotypes using pair-HMM realignment... Running: artic_vcf_filter --medaka my_example.merged.vcf my_example.pass.vcf my_example.fail.vcf Running: bgzip -f my_example.pass.vcf Running: tabix -p vcf my_example.pass.vcf.gz Running: artic_make_depth_mask --store-rg-depths ./primer-schemes/scov2/V3/scov2.reference.fasta my_example.primertrimmed.rg.sorted.bam my_example.coverage_mask.txt Running: artic_mask ./primer-schemes/scov2/V3/scov2.reference.fasta my_example.coverage_mask.txt my_example.fail.vcf my_example.preconsensus.fasta Running: bcftools consensus -f my_example.preconsensus.fasta my_example.pass.vcf.gz -m my_example.coverage_mask.txt -o my_example.consensus.fasta Note: the --sample option not given, applying all records regardless of the genotype Applied 9 variants Running: artic_fasta_header my_example.consensus.fasta "my_example/ARTIC/medaka" Running: cat my_example.consensus.fasta ./primer-schemes/scov2/V3/scov2.reference.fasta > my_example.muscle.in.fasta Running: muscle -in my_example.muscle.in.fasta -out my_example.muscle.out.fasta MUSCLE v3.8.1551 by Robert C. Edgar http://www.drive5.com/muscle This software is donated to the public domain. Please cite: Edgar, R.C. Nucleic Acids Res 32(5), 1792-97. my_example.muscle.in 2 seqs, lengths min 29903, max 29903, avg 29903 00:00:00 2 MB(0%) Iter 1 100.00% K-mer dist pass 1 00:00:00 2 MB(0%) Iter 1 100.00% K-mer dist pass 2 00:00:15 970 MB(6%) Iter 1 100.00% Align node 00:00:15 970 MB(6%) Iter 1 100.00% Root alignment Running: artic_get_stats --scheme ./primer-schemes/scov2/V3/scov2.scheme.bed --align-report my_example.alignreport.txt --vcf-report my_example.vcfreport.txt my_example
That's it! Let's have a quick run through of the parameters we used so that we can understand what was happening.
parameter | explanation |
---|---|
normalise |
This caps amplicon coverage to 200 reads, used mainly to speed up the pipeline run. |
threads |
This sets the number of CPU threads to use during the pipeline. We set this to 2 here as that is the limit on Binder, but if you are playing along at home you can increase this to make things run a bit more quickly. |
medaka |
This tells the ARTIC pipeline to use the medaka workflow |
medaka-model |
This specifies which model to use for the medaka program calls. |
strict |
This runs an additional filtering of reported variants, checking them in overlap regions of the primer scheme to see if they are artifacts reported in only one primer pool. |
read-file |
This tells the pipeline where to find the reads. |
scov/V3 |
This specifies the name of the primer scheme and the version to use. If it isn't found locally, the pipeline will try finding it in the ARTIC primer scheme repository. |
my_exmple |
The name to give this pipeline run, all output will have this prepended to the filenames. |
Now it is time to check what the pipeline has produced for us. The files we are most interested in are:
filename | description |
---|---|
my_example.trimmed.rg.sorted.bam |
the post-processed alignment of reads to the reference genome. |
my_example.primertrimmed.rg.sorted.bam |
the post-processed alignment with additional softmasking to exclude primer sequences. |
my_example.vcfreport.txt |
a report evaluating reported variants against the primer scheme. |
my_example.pass.vcf.gz |
detected variants that passed filters. |
my_example.consensus.fasta |
the consensus sequence for the input sample. |
my_example..muscle.out.fasta |
an alignment of the consensus sequence against the reference sequence. |
Before we look at these files, we can use MultiQC to check our amplicon coverage (pre-normalisation to 100) and if we have any variants reported that may be a result of contamination. Use the ARTIC MultiQC plugin:
!multiqc .
[INFO ] multiqc : This is MultiQC v1.9 [INFO ] multiqc : Template : default [INFO ] multiqc : Searching : /Users/willrowe/Desktop/artic-pipeline-example/notebooks Searching 37 files.. [####################################] 100% [INFO ] custom_content : custom_data_lineplot: Found 1 samples (linegraph) [INFO ] custom_content : custom_data_json_table: Found 1 samples (table) [INFO ] multiqc : Compressing plot data [INFO ] multiqc : Report : multiqc_report.html [INFO ] multiqc : Data : multiqc_data [INFO ] multiqc : MultiQC complete
This will have produced a report HTML file called multiqc_report.html. You can click on that link or use the following code to view it in this notebook:
from IPython.display import IFrame
IFrame(src='./multiqc_report.html', width=600, height=400)
Use the report to see if any amplicons are marked as low coverage due to insufficient reads being assigned. Also use the report to see if there are any overlap variant fails. This is when a variant is idenified within an amplicon overlap region of the scheme but is found in only one amplicon.
As mentioned above, the file containing the filtered variants is my_example.pass.vcf.gz
. Let's take a look:
# import pyVCF and open the variant file
import vcf
vcf_reader = vcf.Reader(filename="my_example.pass.vcf.gz")
# print the variants
print("chromosome\tpos\tref\talt")
for record in vcf_reader:
print("{}\t{}\t{}\t{}" .format(record.CHROM, record.POS, record.REF, record.ALT))
chromosome pos ref alt MN908947.3 2618 A [G] MN908947.3 8782 C [T] MN908947.3 18488 T [C] MN908947.3 21846 C [T] MN908947.3 23605 T [G] MN908947.3 26354 T [A] MN908947.3 28144 T [C] MN908947.3 29366 C [T] MN908947.3 29596 A [G]
We also have the consensus sequence and the alignment files to look out. Here is an example of how we can look at them with Python:
from Bio import AlignIO
alignment = AlignIO.read("my_example.muscle.out.fasta", "fasta")
print("alignment length= {}\n" .format(alignment.get_alignment_length()))
for record in alignment:
print("{}\n{}".format(record.id, record.seq))
alignment length= 29903 my_example/ARTIC/medaka NNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAACTAATTACTGTCGTTGACAGGACACGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAGCACATCTAGGTTTCGTCCGGGTGTGACCGAAAGGTAAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGAAAACACACGTCCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGGCACGTCAACATCTTAAAGATGGCACTTGTGGCTTAGTAGAAGTTGAAAAAGGCGTTTTGCCTCAACTTGAACAGCCCTATGTGTTCATCAAACGTTCGGATGCTCGAACTGCACCTCATGGTCATGTTATGGTTGAGCTGGTAGCAGAACTCGAAGGCATTCAGTACGGTCGTAGTGGTGAGACACTTGGTGTCCTTGTCCCTCATGTGGGCGAAATACCAGTGGCTTACCGCAAGGTTCTTCTTCGTAAGAACGGTAATAAAGGAGCTGGTGGCCATAGTTACGGCGCCGATCTAAAGTCATTTGACTTAGGCGACGAGCTTGGCACTGATCCTTATGAAGATTTTCAAGAAAACTGGAACACTAAACATAGCAGTGGTGTTACCCGTGAACTCATGCGTGAGCTTAACGGAGGGGCATACACTCGCTATGTCGATAACAACTTCTGTGGCCCTGATGGCTACCCTCTTGAGTGCATTAAAGACCTTCTAGCACGTGCTGGTAAAGCTTCATGCACTTTGTCCGAACAACTGGACTTTATTGACACTAAGAGGGGTGTATACTGCTGCCGTGAACATGAGCATGAAATTGCTTGGTACACGGAACGTTCTGAAAAGAGCTATGAATTGCAGACACCTTTTGAAATTAAATTGGCAAAGAAATTTGACACCTTCAATGGGGAATGTCCAAATTTTGTATTTCCCTTAAATTCCATAATCAAGACTATTCAACCAAGGGTTGAAAAGAAAAAGCTTGATGGCTTTATGGGTAGAATTCGATCTGTCTATCCAGTTGCGTCACCAAATGAATGCAACCAAATGTGCCTTTCAACTCTCATGAAGTGTGATCATTGTGGTGAAACTTCATGGCAGACGGGCGATTTTGTTAAAGCCACTTGCGAATTTTGTGGCACTGAGAATTTGACTAAAGAAGGTGCCACTACTTGTGGTTACTTACCCCAAAATGCTGTTGTTAAAATTTATTGTCCAGCATGTCACAATTCAGAAGTAGGACCTGAGCATAGTCTTGCCGAATACCATAATGAATCTGGCTTGAAAACCATTCTTCGTAAGGGTGGTCGCACTATTGCCTTTGGAGGCTGTGTGTTCTCTTATGTTGGTTGCCATAACAAGTGTGCCTATTGGGTTCCACGTGCTAGCGCTAACATAGGTTGTAACCATACAGGTGTTGTTGGAGAAGGTTCCGAAGGTCTTAATGACAACCTTCTTGAAATACTCCAAAAAGAGAAAGTCAACATCAATATTGTTGGTGACTTTAAACTTAATGAAGAGATCGCCATTATTTTGGCATCTTTTTCTGCTTCCACAAGTGCTTTTGTGGAAACTGTGAAAGGTTTGGATTATAAAGCATTCAAACAAATTGTTGAATCCTGTGGTAATTTTAAAGTTACAAAAGGAAAAGCTAAAAAAGGTGCCTGGAATATTGGTGAACAGAAATCAATACTGAGTCCTCTTTATGCATTTGCATCAGAGGCTGCTCGTGTTGTACGATCAATTTTCTCCCGCACTCTTGAAACTGCTCAAAATTCTGTGCGTGTTTTACAGAAGGCCGCTATAACAATACTAGATGGAATTTCACAGTATTCACTGAGACTCATTGATGCTATGATGTTCACATCTGATTTGGCTACTAACAATCTAGTTGTAATGGCCTACATTACAGGTGGTGTTGTTCAGTTGACTTCGCAGTGGCTAACTAACATCTTTGGCACTGTTTATGAAAAACTCAAACCCGTCCTTGATTGGCTTGAAGAGAAGTTTAAGGAAGGTGTAGAGTTTCTTAGAGACGGTTGGGAAATTGTTAAATTTATCTCAACCTGTGCTTGTGAAATTGTCGGTGGACAAATTGTCACCTGTGCAAAGGAAATTAAGGAGAGTGTTCAGACATTCTTTAAGCTTGTAAATAAATTTTTGGCTTTGTGTGCTGACTCTATCATTATTGGTGGAGCTAAACTTAAAGCCTTGAATTTAGGTGAAACATTTGTCACGCACTCAAAGGGATTGTACAGAAAGTGTGTTAAATCCAGAGAAGAAACTGGCCTACTCATGCCTCTAAAAGCCCCAAAAGAAATTATCTTCTTAGAGGGAGAAACACTTCCCACAGAAGTGTTAACAGAGGAAGTTGTCTTGAAAACTGGTGATTTACAACCATTAGAACAACCTACTAGTGAAGCTGTTGAAGCTCCATTGGTTGGTACACCAGTTTGTGTTAACGGGCTTATGTTGCTCGAAATCAAAGACACAGAAAAGTACTGTGCCCTTGCACCTAATATGATGGTAACAAACAATACCTTCACACTCAAAGGCGGTGCACCAACAAAGGTTACTTTTGGTGATGACACTGTGATAGAAGTGCAAGGTTACAAGAGTGTGAATATCACTTTTGAACTTGATGAAAGGATTGATAAAGTACTTAATGAGAAGTGCTCTGCCTATACAGTTGAACTCGGTACAGAAGTAAATGAGTTCGCCTGTGTTGTGGCAGATGCTGTCATAAAAACTTTGCAACCAGTATCTGAATTACTTACACCACTGGGCATTGATTTAGATGAGTGGAGTATGGCTACATACTACTTATTTGATGAGTCTGGTGAGTTTAAATTGGCTTCACATATGTATTGTTCTTTCTACCCTCCAGATGAGGATGAAGAAGAAGGTGATTGTGAAGAAGAAGAGTTTGAGCCATCAACTCAATATGAGTATGGTACTGAAGATGATTACCAAGGTAAACCTTTGGAATTTGGTGCCACTTCTGCTGCTCTTCAACCTGAAGAAGAGCAAGAAGAAGATTGGTTAGATGATGATAGTCAACAAACTGTTGGTCAACAAGACGGCAGTGAGGACAATCAGACAACTACTATTCAAACAATTGTTGAGGTTCAACCTCAATTAGAGATGGAACTTACACCAGTTGTTCAGACTATTGAAGTGAATAGTTTTAGTGGTTATTTAAAACTTACTGACAATGTATACATTAAAAATGCAGACATTGTGGAAGAAGCTAAAAAGGTAAAACCAACAGTGGTTGTTAATGCAGCCAATGTTTACCTTAAACATGGAGGAGGTGTTGCAGGAGCCTTAAATAAGGCTACTAACAATGCCATGCAAGTTGAATCTGATGATTACATAGCTACTAATGGACCACTTAAAGTGGGTGGTAGTTGTGTTTTAAGCGGACACAATCTTGCTAAACACTGTCTTCATGTTGTCGGCCCAAATGTTAACAAAGGTGAAGACATTCAACTTCTTAAGAGTGCTTATGAAAATTTTAATCAGCACGAAGTTCTACTTGCACCATTATTATCAGCTGGTATTTTTGGTGCTGACCCTATACATTCTTTAAGAGTTTGTGTAGATACTGTTCGCACAAATGTCTACTTAGCTGTCTTTGATAAAAATCTCTATGACAAACTTGTTTCAAGCTTTTTGGAAATGAAGAGTGAAAAGCAAGTTGAACAAAAGATCGCTGAGATTCCTAAAGAGGAAGTTAAGCCATTTATAACTGAAAGTAAACCTTCAGTTGAACAGAGAAAACAAGATGATAAGAAAATCAAAGCTTGTGTTGAAGAAGTTACAACAACTCTGGAAGAAACTAAGTTCCTCACAGAAAACTTGTTACTTTATATTGACATTAATGGCAATCTTCATCCAGATTCTGCCACTCTTGTTAGTGACATTGACATCACTTTCTTAAAGAAAGATGCTCCATATATAGTGGGTGATGTTGTTCAAGAGGGTGTTTTAACTGCTGTGGTTATACCTACTAAAAAGGCTGGTGGCACTACTGAAATGCTAGCGAAAGCTTTGAGAAAAGTGCCAACAGACAATTATATAACCACTTACCCGGGTCAGGGTTTAAATGGTTACACTGTAGAGGAGGCAAAGACAGTGCTTAAAAAGTGTAAAAGTGCCTTTTACATTCTACCATCTATTATCTCTAATGAGAAGCAAGAAATTCTTGGAACTGTTTCTTGGAATTTGCGAGAAATGCTTGCACATGCAGAAGAAACACGCAAATTAATGCCTGTCTGTGTGGAAACTAAAGCCATAGTTTCAACTATACAGCGTAAATATAAGGGTATTAAAATACAAGAGGGTGTGGTTGATTATGGTGCTAGATTTTACTTTTACACCAGTAAAACAACTGTAGCGTCACTTATCAACACACTTAACGATCTAAATGAAACTCTTGTTACAATGCCACTTGGCTATGTAACACATGGCTTAAATTTGGAAGAAGCTGCTCGGTATATGAGATCTCTCAAAGTGCCAGCTACAGTTTCTGTTTCTTCACCTGATGCTGTTACAGCGTATAATGGTTATCTTACTTCTTCTTCTAAAACACCTGAAGAACATTTTATTGAAACCATCTCACTTGCTGGTTCCTATAAAGATTGGTCCTATTCTGGACAATCTACACAACTAGGTATAGAATTTCTTAAGAGAGGTGATAAAAGTGTATATTACACTAGTAATCCTACCACATTCCACCTAGATGGTGAAGTTATCACCTTTGACAATCTTAAGACACTTCTTTCTTTGAGAGAAGTGAGGACTATTAAGGTGTTTACAACAGTAGACAACATTAACCTCCACACGCAAGTTGTGGACATGTCAATGACATATGGACAACAGTTTGGTCCAACTTATTTGGATGGAGCTGATGTTACTAAAATAAAACCTCATAATTCACATGAAGGTAAAACATTTTATGTTTTACCTAATGATGACACTCTACGTGTTGAGGCTTTTGAGTACTACCACACAACTGATCCTAGTTTTCTGGGTAGGTACATGTCAGCATTAAATCACACTAAAAAGTGGAAATACCCACAAGTTAATGGTTTAACTTCTATTAAATGGGCAGATAACAACTGTTATCTTGCCACTGCATTGTTAACACTCCAACAAATAGAGTTGAAGTTTAATCCACCTGCTCTACAAGATGCTTATTACAGAGCAAGGGCTGGTGAAGCTGCTAACTTTTGTGCACTTATCTTAGCCTACTGTAATAAGACAGTAGGTGAGTTAGGTGATGTTAGAGAAACAATGAGTTACTTGTTTCAACATGCCAATTTAGATTCTTGCAAAAGAGTCTTGAACGTGGTGTGTAAAACTTGTGGACAACAGCAGACAACCCTTAAGGGTGTAGAAGCTGTTATGTACATGGGCACACTTTCTTATGAACAATTTAAGAAAGGTGTTCAGATACCTTGTACGTGTGGTAAACAAGCTACAAAATATCTAGTACAACAGGAGTCACCTTTTGTTATGATGTCAGCACCACCTGCTCAGTATGAACTTAAGCATGGTACATTTACTTGTGCTAGTGAGTACACTGGTAATTACCAGTGTGGTCACTATAAACATATAACTTCTAAAGAAACTTTGTATTGCATAGACGGTGCTTTACTTACAAAGTCCTCAGAATACAAAGGTCCTATTACGGATGTTTTCTACAAAGAAAACAGTTACACAACAACCATAAAACCAGTTACTTATAAATTGGATGGTGTTGTTTGTACAGAAATTGACCCTAAGTTGGACAATTATTATAAGAAAGACAATTCTTATTTCACAGAGCAACCAATTGATCTTGTACCAAACCAACCATATCCAAACGCAAGCTTCGATAATTTTAAGTTTGTATGTGATAATATCAAATTTGCTGATGATTTAAACCAGTTAACTGGTTATAAGAAACCTGCTTCAAGAGAGCTTAAAGTTACATTTTTCCCTGACTTAAATGGTGATGTGGTGGCTATTGATTATAAACACTACACACCCTCTTTTAAGAAAGGAGCTAAATTGTTACATAAACCTATTGTTTGGCATGTTAACAATGCAACTAATAAAGCCACGTATAAACCAAATACCTGGTGTATACGTTGTCTTTGGAGCACAAAACCAGTTGAAACATCAAATTCGTTTGATGTACTGAAGTCAGAGGACGCGCAGGGAATGGATAATCTTGCCTGCGAAGATCTAAAACCAGTCTCTGAAGAAGTAGTGGAAAATCCTACCATACAGAAAGACGTTCTTGAGTGTAATGTGAAAACTACCGAAGTTGTAGGAGACATTATACTTAAACCAGCAAATAATAGTTTAAAAATTACAGAAGAGGTTGGCCACACAGATCTAATGGCTGCTTATGTAGACAATTCTAGTCTTACTATTAAGAAACCTAATGAATTATCTAGAGTATTAGGTTTGAAAACCCTTGCTACTCATGGTTTAGCTGCTGTTAATAGTGTCCCTTGGGATACTATAGCTAATTATGCTAAGCCTTTTCTTAACAAAGTTGTTAGTACAACTACTAACATAGTTACACGGTGTTTAAACCGTGTTTGTACTAATTATATGCCTTATTTCTTTACTTTATTGCTACAATTGTGTACTTTTACTAGAAGTACAAATTCTAGAATTAAAGCATCTATGCCGACTACTATAGCAAAGAATACTGTTAAGAGTGTCGGTAAATTTTGTCTAGAGGCTTCATTTAATTATTTGAAGTCACCTAATTTTTCTAAACTGATAAATATTATAATTTGGTTTTTACTATTAAGTGTTTGCCTAGGTTCTTTAATCTACTCAACCGCTGCTTTAGGTGTTTTAATGTCTAATTTAGGCATGCCTTCTTACTGTACTGGTTACAGAGAAGGCTATTTGAACTCTACTAATGTCACTATTGCAACCTACTGTACTGGTTCTATACCTTGTAGTGTTTGTCTTAGTGGTTTAGATTCTTTAGACACCTATCCTTCTTTAGAAACTATACAAATTACCATTTCATCTTTTAAATGGGATTTAACTGCTTTTGGCTTAGTTGCAGAGTGGTTTTTGGCATATATTCTTTTCACTAGGTTTTTCTATGTACTTGGATTGGCTGCAATCATGCAATTGTTTTTCAGCTATTTTGCAGTACATTTTATTAGTAATTCTTGGCTTATGTGGTTAATAATTAATCTTGTACAAATGGCCCCGATTTCAGCTATGGTTAGAATGTACATCTTCTTTGCATCATTTTATTATGTATGGAAAAGTTATGTGCATGTTGTAGACGGTTGTAATTCATCAACTTGTATGATGTGTTACAAACGTAATAGAGCAACAAGAGTCGAATGTACAACTATTGTTAATGGTGTTAGAAGGTCCTTTTATGTCTATGCTAATGGAGGTAAAGGCTTTTGCAAACTACACAATTGGAATTGTGTTAATTGTGATACATTCTGTGCTGGTAGTACATTTATTAGTGATGAAGTTGCGAGAGACTTGTCACTACAGTTTAAAAGACCAATAAATCCTACTGACCAGTCTTCTTACATCGTTGATAGTGTTACAGTGAAGAATGGTTCCATCCATCTTTACTTTGATAAAGCTGGTCAAAAGACTTATGAAAGACATTCTCTCTCTCATTTTGTTAACTTAGACAACCTGAGAGCTAATAACACTAAAGGTTCATTGCCTATTAATGTTATAGTTTTTGATGGTAAATCAAAATGTGAAGAATCATCTGCAAAATCAGCGTCTGTTTACTACAGTCAGCTTATGTGTCAACCTATACTGTTACTAGATCAGGCATTAGTGTCTGATGTTGGTGATAGTGCGGAAGTTGCAGTTAAAATGTTTGATGCTTACGTTAATACGTTTTCATCAACTTTTAACGTACCAATGGAAAAACTCAAAACACTAGTTGCAACTGCAGAAGCTGAACTTGCAAAGAATGTGTCCTTAGACAATGTCTTATCTACTTTTATTTCAGCAGCTCGGCAAGGGTTTGTTGATTCAGATGTAGAAACTAAAGATGTTGTTGAATGTCTTAAATTGTCACATCAATCTGACATAGAAGTTACTGGCGATAGTTGTAATAACTATATGCTCACCTATAACAAAGTTGAAAACATGACACCCCGTGACCTTGGTGCTTGTATTGACTGTAGTGCGCGTCATATTAATGCGCAGGTAGCAAAAAGTCACAACATTGCTTTGATATGGAACGTTAAAGATTTCATGTCATTGTCTGAACAACTACGAAAACAAATACGTAGTGCTGCTAAAAAGAATAACTTACCTTTTAAGTTGACATGTGCAACTACTAGACAAGTTGTTAATGTTGTAACAACAAAGATAGCACTTAAGGGTGGTAAAATTGTTAATAATTGGTTGAAGCAGTTAATTAAAGTTACACTTGTGTTCCTTTTTGTTGCTGCTATTTTCTATTTAATAACACCTGTTCATGTCATGTCTAAACATACTGACTTTTCAAGTGAAATCATAGGATACAAGGCTATTGATGGTGGTGTCACTCGTGACATAGCATCTACAGATACTTGTTTTGCTAACAAACATGCTGATTTTGACACATGGTTTAGTCAGCGTGGTGGTAGTTATACTAATGACAAAGCTTGCCCATTGATTGCTGCAGTCATAACAAGAGAAGTGGGTTTTGTCGTGCCTGGTTTGCCTGGCACGATATTACGCACAACTAATGGTGACTTTTTGCATTTCTTACCTAGAGTTTTTAGTGCAGTTGGTAACATCTGTTACACACCATCAAAACTTATAGAGTACACTGACTTTGCAACATCAGCTTGTGTTTTGGCTGCTGAATGTACAATTTTTAAAGATGCTTCTGGTAAGCCAGTACCATATTGTTATGATACCAATGTACTAGAAGGTTCTGTTGCTTATGAAAGTTTACGCCCTGACACACGTTATGTGCTCATGGATGGCTCTATTATTCAATTTCCTAACACCTACCTTGAAGGTTCTGTTAGAGTGGTAACAACTTTTGATTCTGAGTACTGTAGGCACGGCACTTGTGAAAGATCAGAAGCTGGTGTTTGTGTATCTACTAGTGGTAGATGGGTACTTAACAATGATTATTACAGATCTTTACCAGGAGTTTTCTGTGGTGTAGATGCTGTAAATTTACTTACTAATATGTTTACACCACTAATTCAACCTATTGGTGCTTTGGACATATCAGCATCTATAGTAGCTGGTGGTATTGTAGCTATCGTAGTAACATGCCTTGCCTACTATTTTATGAGGTTTAGAAGAGCTTTTGGTGAATACAGTCATGTAGTTGCCTTTAATACTTTACTATTCCTTATGTCATTCACTGTACTCTGTTTAACACCAGTTTACTCATTCTTACCTGGTGTTTATTCTGTTATTTACTTGTACTTGACATTTTATCTTACTAATGATGTTTCTTTTTTAGCACATATTCAGTGGATGGTTATGTTCACACCTTTAGTACCTTTCTGGATAACAATTGCTTATATCATTTGTATTTCCACAAAGCATTTCTATTGGTTCTTTAGTAATTACCTAAAGAGACGTGTAGTCTTTAATGGTGTTTCCTTTAGTACTTTTGAAGAAGCTGCGCTGTGCACCTTTTTGTTAAATAAAGAAATGTATCTAAAGTTGCGTAGTGATGTGCTATTACCTCTTACGCAATATAATAGATACTTAGCTCTTTATAATAAGTACAAGTATTTTAGTGGAGCAATGGATACAACTAGCTACAGAGAAGCTGCTTGTTGTCATCTCGCAAAGGCTCTCAATGACTTCAGTAACTCAGGTTCTGATGTTCTTTACCAACCACCACAAACCTCTATCACCTCAGCTGTTTTGCAGAGTGGTTTTAGAAAAATGGCATTCCCATCTGGTAAAGTTGAGGGTTGTATGGTACAAGTAACTTGTGGTACAACTACACTTAACGGTCTTTGGCTTGATGACGTAGTTTACTGTCCAAGACATGTGATCTGCACCTCTGAAGACATGCTTAACCCTAATTATGAAGATTTACTCATTCGTAAGTCTAATCATAATTTCTTGGTACAGGCTGGTAATGTTCAACTCAGGGTTATTGGACATTCTATGCAAAATTGTGTACTTAAGCTTAAGGTTGATACAGCCAATCCTAAGACACCTAAGTATAAGTTTGTTCGCATTCAACCAGGACAGACTTTTTCAGTGTTAGCTTGTTACAATGGTTCACCATCTGGTGTTTACCAATGTGCTATGAGGCCCAATTTCACTATTAAGGGTTCATTCCTTAATGGTTCATGTGGTAGTGTTGGTTTTAACATAGATTATGACTGTGTCTCTTTTTGTTACATGCACCATATGGAATTACCAACTGGAGTTCATGCTGGCACAGACTTAGAAGGTAACTTTTATGGACCTTTTGTTGACAGGCAAACAGCACAAGCAGCTGGTACGGACACAACTATTACAGTTAATGTTTTAGCTTGGTTGTACGCTGCTGTTATAAATGGAGACAGGTGGTTTCTCAATCGATTTACCACAACTCTTAATGACTTTAACCTTGTGGCTATGAAGTACAATTATGAACCTCTAACACAAGACCATGTTGACATACTAGGACCTCTTTCTGCTCAAACTGGAATTGCCGTTTTAGATATGTGTGCTTCATTAAAAGAATTACTGCAAAATGGTATGAATGGACGTACCATATTGGGTAGTGCTTTATTAGAAGATGAATTTACACCTTTTGATGTTGTTAGACAATGCTCAGGTGTTACTTTCCAAAGTGCAGTGAAAAGAACAATCAAGGGTACACACCACTGGTTGTTACTCACAATTTTGACTTCACTTTTAGTTTTAGTCCAGAGTACTCAATGGTCTTTGTTCTTTTTTTTGTATGAAAATGCCTTTTTACCTTTTGCTATGGGTATTATTGCTATGTCTGCTTTTGCAATGATGTTTGTCAAACATAAGCATGCATTTCTCTGTTTGTTTTTGTTACCTTCTCTTGCCACTGTAGCTTATTTTAATATGGTCTATATGCCTGCTAGTTGGGTGATGCGTATTATGACATGGTTGGATATGGTTGATACTAGTTTGTCTGGTTTTAAGCTAAAAGACTGTGTTATGTATGCATCAGCTGTAGTGTTACTAATCCTTATGACAGCAAGAACTGTGTATGATGATGGTGCTAGGAGAGTGTGGACACTTATGAATGTCTTGACACTCGTTTATAAAGTTTATTATGGTAATGCTTTAGATCAAGCCATTTCCATGTGGGCTCTTATAATCTCTGTTACTTCTAACTACTCAGGTGTAGTTACAACTGTCATGTTTTTGGCCAGAGGTATTGTTTTTATGTGTGTTGAGTATTGCCCTATTTTCTTCATAACTGGTAATACACTTCAGTGTATAATGCTAGTTTATTGTTTCTTAGGCTATTTTTGTACTTGTTACTTTGGCCTCTTTTGTTTACTCAACCGCTACTTTAGACTGACTCTTGGTGTTTATGATTACTTAGTTTCTACACAGGAGTTTAGATATATGAATTCACAGGGACTACTCCCACCCAAGAATAGCATAGATGCCTTCAAACTCAACATTAAATTGTTGGGTGTTGGTGGCAAACCTTGTATCAAAGTAGCCACTGTACAGTCTAAAATGTCAGATGTAAAGTGCACATCAGTAGTCTTACTCTCAGTTTTGCAACAACTCAGAGTAGAATCATCATCTAAATTGTGGGCTCAATGTGTCCAGTTACACAATGACATTCTCTTAGCTAAAGATACTACTGAAGCCTTTGAAAAAATGGTTTCACTACTTTCTGTTTTGCTTTCCATGCAGGGTGCTGTAGACATAAACAAGCTTTGTGAAGAAATGCTGGACAACAGGGCAACCTTACAAGCTATAGCCTCAGAGTTTAGTTCCCTTCCATCATATGCAGCTTTTGCTACTGCTCAAGAAGCTTATGAGCAGGCTGTTGCTAATGGTGATTCTGAAGTTGTTCTTAAAAAGTTGAAGAAGTCTTTGAATGTGGCTAAATCTGAATTTGACCGTGATGCAGCCATGCAACGTAAGTTGGAAAAGATGGCTGATCAAGCTATGACCCAAATGTATAAACAGGCTAGATCTGAGGACAAGAGGGCAAAAGTTACTAGTGCTATGCAGACAATGCTTTTCACTATGCTTAGAAAGTTGGATAATGATGCACTCAACAACATTATCAACAATGCAAGAGATGGTTGTGTTCCCTTGAACATAATACCTCTTACAACAGCAGCCAAACTAATGGTTGTCATACCAGACTATAACACATATAAAAATACGTGTGATGGTACAACATTTACTTATGCATCAGCATTGTGGGAAATCCAACAGGTTGTAGATGCAGATAGTAAAATTGTTCAACTTAGTGAAATTAGTATGGACAATTCACCTAATTTAGCATGGCCTCTTATTGTAACAGCTTTAAGGGCCAATTCTGCTGTCAAATTACAGAATAATGAGCTTAGTCCTGTTGCACTACGACAGATGTCTTGTGCTGCCGGTACTACACAAACTGCTTGCACTGATGACAATGCGTTAGCTTACTACAACACAACAAAGGGAGGTAGGTTTGTACTTGCACTGTTATCCGATTTACAGGATTTGAAATGGGCTAGATTCCCTAAGAGTGATGGAACTGGTACTATCTATACAGAACTGGAACCACCTTGTAGGTTTGTTACAGACACACCTAAAGGTCCTAAAGTGAAGTATTTATACTTTATTAAAGGATTAAACAACCTAAATAGAGGTATGGTACTTGGTAGTTTAGCTGCCACAGTACGTCTACAAGCTGGTAATGCAACAGAAGTGCCTGCCAATTCAACTGTATTATCTTTCTGTGCTTTTGCTGTAGATGCTGCTAAAGCTTACAAAGATTATCTAGCTAGTGGGGGACAACCAATCACTAATTGTGTTAAGATGTTGTGTACACACACTGGTACTGGTCAGGCAATAACAGTTACACCGGAAGCCAATATGGATCAAGAATCCTTTGGTGGTGCATCGTGTTGTCTGTACTGCCGTTGCCACATAGATCATCCAAATCCTAAAGGATTTTGTGACTTAAAAGGTAAGTATGTACAAATACCTACAACTTGTGCTAATGACCCTGTGGGTTTTACACTTAAAAACACAGTCTGTACCGTCTGCGGTATGTGGAAAGGTTATGGCTGTAGTTGTGATCAACTCCGCGAACCCATGCTTCAGTCAGCTGATGCACAATCGTTTTTAAACGGGTTTGCGGTGTAAGTGCAGCCCGTCTTACACCGTGCGGCACAGGCACTAGTACTGATGTCGTATACAGGGCTTTTGACATCTACAATGATAAAGTAGCTGGTTTTGCTAAATTCCTAAAAACTAATTGTTGTCGCTTCCAAGAAAAGGACGAAGATGACAATTTAATTGATTCTTACTTTGTAGTTAAGAGACACACTTTCTCTAACTACCAACATGAAGAAACAATTTATAATTTACTTAAGGATTGTCCAGCTGTTGCTAAACATGACTTCTTTAAGTTTAGAATAGACGGTGACATGGTACCACATATATCACGTCAACGTCTTACTAAATACACAATGGCAGACCTCGTCTATGCTTTAAGGCATTTTGATGAAGGTAATTGTGACACATTAAAAGAAATACTTGTCACATACAATTGTTGTGATGATGATTATTTCAATAAAAAGGACTGGTATGATTTTGTAGAAAACCCAGATATATTACGCGTATACGCCAACTTAGGTGAACGTGTACGCCAAGCTTTGTTAAAAACAGTACAATTCTGTGATGCCATGCGAAATGCTGGTATTGTTGGTGTACTGACATTAGATAATCAAGATCTCAATGGTAACTGGTATGATTTCGGTGATTTCATACAAACCACGCCAGGTAGTGGAGTTCCTGTTGTAGATTCTTATTATTCATTGTTAATGCCTATATTAACCTTGACCAGGGCTTTAACTGCAGAGTCACATGTTGACACTGACTTAACAAAGCCTTACATTAAGTGGGATTTGTTAAAATATGACTTCACGGAAGAGAGGTTAAAACTCTTTGACCGTTATTTTAAATATTGGGATCAGACATACCACCCAAATTGTGTTAACTGTTTGGATGACAGATGCATTCTGCATTGTGCAAACTTTAATGTTTTATTCTCTACAGTGTTCCCACCTACAAGTTTTGGACCACTAGTGAGAAAAATATTTGTTGATGGTGTTCCATTTGTAGTTTCAACTGGATACCACTTCAGAGAGCTAGGTGTTGTACATAATCAGGATGTAAACTTACATAGCTCTAGACTTAGTTTTAAGGAATTACTTGTGTATGCTGCTGACCCTGCTATGCACGCTGCTTCTGGTAATCTATTACTAGATAAACGCACTACGTGCTTTTCAGTAGCTGCACTTACTAACAATGTTGCTTTTCAAACTGTCAAACCCGGTAATTTTAACAAAGACTTCTATGACTTTGCTGTGTCTAAGGGTTTCTTTAAGGAAGGAAGTTCTGTTGAATTAAAACACTTCTTCTTTGCTCAGGATGGTAATGCTGCTATCAGCGATTATGACTACTATCGTTATAATCTACCAACAATGTGTGATATCAGACAACTACTATTTGTAGTTGAAGTTGTTGATAAGTACTTTGATTGTTACGATGGTGGCTGTATTAATGCTAACCAAGTCATCGTCAACAACCTAGACAAATCAGCTGGTTTTCCATTTAATAAATGGGGTAAGGCTAGACTTTATTATGATTCAATGAGTTATGAGGATCAAGATGCACTTTTCGCATATACAAAACGTAATGTCATCCCTACTATAACTCAAATGAATCTTAAGTATGCCATTAGTGCAAAGAATAGAGCTCGCACCGTAGCTGGTGTCTCTATCTGTAGTACTATGACCAATAGACAGTTTCATCAAAAATTATTGAAATCAATAGCCGCCACTAGAGGAGCTACTGTAGTAATTGGAACAAGCAAATTCTATGGTGGTTGGCACAACATGTTAAAAACTGTTTATAGTGATGTAGAAAACCCTCACCTTATGGGTTGGGATTATCCTAAATGTGATAGAGCCATGCCTAACATGCTTAGAATTATGGCCTCACTTGTTCTTGCTCGCAAACATACAACGTGTTGTAGCTTGTCACACCGTTTCTATAGATTAGCTAATGAGTGTGCTCAAGTATTGAGTGAAATGGTCATGTGTGGCGGTTCACTATATGTTAAACCAGGTGGAACCTCATCAGGAGATGCCACAACTGCTTATGCTAATAGTGTTTTTAACATTTGTCAAGCTGTCACGGCCAATGTTAATGCACTTTTATCTACTGATGGTAACAAAATTGCCGATAAGTATGTCCGCAATTTACAACACAGACTTTATGAGTGTCTCTATAGAAATAGAGATGTTGACACAGACTTTGTGAATGAGTTTTACGCATATTTGCGTAAACATTTCTCAATGATGATACTCTCTGACGATGCTGTTGTGTGTTTCAATAGCACTTATGCATCTCAAGGTCTAGTGGCTAGCATAAAGAACTTTAAGTCAGTTCTTTATTATCAAAACAATGTTTTTATGTCTGAAGCAAAATGTTGGACTGAGACTGACCTTACTAAAGGACCTCATGAATTTTGCTCTCAACATACAATGCTAGTTAAACAGGGTGATGATTATGTGTACCTTCCTTACCCAGATCCATCAAGAATCCTAGGGGCCGGCTGTTTTGTAGATGATATCGTAAAAACAGATGGTACACTTATGATTGAACGGTTCGTGTCTTTAGCTATAGATGCTTACCCACTTACTAAACATCCTAATCAGGAGTATGCTGATGTCTTTCATTTGTACTTACAATACATAAGAAAGCTACATGATGAGTTAACAGGACACATGTTAGACATGTATTCTGTTATGCTTACTAATGATAACACTTCAAGGTATTGGGAACCTGAGTTTTATGAGGCTATGTACACACCGCATACAGTCTTACAGGCTGTTGGGGCTTGTGTTCTTTGCAATTCACAGACTTCATTAAGATGTGGTGCTTGCATACGTAGACCATTCTTATGTTGTAAATGCTGTTACGACCATGTCATATCAACATCACATAAATTAGTCTTGTCTGTTAATCCGTATGTTTGCAATGCTCCAGGTTGTGATGTCACAGATGTGACTCAACTTTACTTAGGAGGTATGAGCTATTATTGTAAATCACATAAACCACCCATTAGTTTTCCATTGTGTGCTAATGGACAAGTTTTTGGTTTATATAAAAATACATGTGTTGGTAGCGATAATGTTACTGACTTTAATGCAATTGCAACATGTGACTGGACAAATGCTGGTGATTACATTTTAGCTAACACCTGTACTGAAAGACTCAAGCTTTTTGCAGCAGAAACGCTCAAAGCTACTGAGGAGACATTTAAACTGTCTTATGGTATTGCTACTGTACGTGAAGTGCTGTCTGACAGAGAATTACATCTTTCATGGGAAGTTGGTAAACCTAGACCACCACTTAACCGAAATTATGTCTTTACTGGTTATCGTGTAACTAAAAACAGTAAAGTACAAATAGGAGAGTACACCTTTGAAAAAGGTGACTATGGTGATGCTGTTGTTTACCGAGGTACAACAACTTACAAATTAAATGTTGGTGATTATTTTGTGCTGACATCACATACAGTAATGCCATTAAGTGCACCTACACTAGTGCCACAAGAGCACTATGTTAGAATTACTGGCTTATACCCAACACTCAATATCTCAGATGAGTTTTCTAGCAATGTTGCAAATTATCAAAAGGTTGGTATGCAAAAGTATTCTACACTCCAGGGACCACCTGGTACTGGTAAGAGTCATTTTGCTATTGGCCTAGCTCTCTACTACCCTTCTGCTCGCATAGTGTATACAGCTTGCTCTCATGCCGCTGTTGATGCACTATGTGAGAAGGCATTAAAATATTTGCCTATAGATAAATGTAGTAGAATTATACCTGCACGTGCTCGTGTAGAGTGTTTTGATAAATTCAAAGTGAATTCAACATTAGAACAGTATGTCTTTTGTACTGTAAATGCATTGCCTGAGACGACAGCAGATATAGTTGTCTTTGATGAAATTTCAATGGCCACAAATTATGATTTGAGTGTTGTCAATGCCAGATTACGTGCTAAGCACTATGTGTACATTGGCGACCCTGCTCAATTACCTGCACCACGCACATTGCTAACTAAGGGCACACTAGAACCAGAATATTTCAATTCAGTGTGTAGACTTATGAAAACTATAGGTCCAGACATGTTCCTCGGAACTTGTCGGCGTTGTCCTGCTGAAATTGTTGACACTGTGAGTGCTTTGGTTTATGATAATAAGCTTAAAGCACATAAAGACAAATCAGCTCAATGCTTTAAAATGTTTTATAAGGGTGTTATCACGCATGATGTTTCATCTGCAATTAACAGGCCACAAATAGGCGTGGTAAGAGAATTCCTTACACGTAACCCTGCTTGGAGAAAAGCTGTCTTTATTTCACCTTATAATTCACAGAATGCTGTAGCCTCAAAGATTTTGGGACTACCAACTCAAACTGTTGATTCATCACAGGGCTCAGAATATGACTATGTCATATTCACTCAAACCACTGAAACAGCTCACTCTTGTAATGTAAACAGATTTAATGTTGCTATTACCAGAGCAAAAGTAGGCATACTTTGCATAATGTCTGATAGAGACCTTTATGACAAGTTGCAATTTACAAGTCTTGAAATTCCACGTAGGAATGTGGCAACTTTACAAGCTGAAAATGTAACAGGACTCTTTAAAGATTGTAGTAAGGTAATCACTGGGTTACATCCTACACAGGCACCTACACACCTCAGTGTTGACACTAAATTCAAAACTGAAGGTTTATGTGTTGACATACCTGGCATACCTAAGGACATGACCTATAGAAGACTCATCTCTATGATGGGTTTTAAAATGAATTATCAAGTTAATGGTTACCCTAACATGTTTATCACCCGCGAAGAAGCTATAAGACATGTACGTGCATGGATTGGCTTCGATGTCGAGGGGTGTCATGCTACTAGAGAAGCTGTTGGTACCAATTTACCTTTACAGCTAGGTTTTTCTACAGGTGTTAACCTAGTTGCTGTACCTACAGGTTATGTTGATACACCTAATAATACAGATTTTTCCAGAGTTAGTGCTAAACCACCGCCTGGAGATCAATTTAAACACCTCACACCACTTATGTACAAAGGACTTCCTTGGAATGTAGTGCGTATAAAGATTGTACAAATGTTAAGTGACACACTTAAAAATCTCTCTGACAGAGTCGTATTTGTCTTATGGGCACATGGCTTTGAGTTGACATCTATGAAGTATTTTGTGAAAATAGGACCTGAGCGCACCTGTTGTCTATGTGATAGACGTGCCACATGCTTTTCCACTGCTTCAGACACTTATGCCTGTTGGCATCATTCTATTGGATTTGATTACGTCTATAATCCGTTTATGATTGATGTTCAACAATGGGGTTTTACAGGTAACCTACAAAGCAACCATGATCTGTATTGTCAAGTCCATGGTAATGCACATGTAGCTAGTTGTGATGCAATCATGACTAGGTGTCTAGCTGTCCACGAGTGCTTTGTTAAGCGTGTTGACTGGACTATTGAATATCCTATAATTGGTGATGAACTGAAGATTAATGCGGCTTGTAGAAAGGTTCAACACATGGTTGTTAAAGCTGCATTATTAGCAGACAAATTCCCAGTTCTTCACGACATTGGTAACCCTAAAGCTATTAAGTGTGTACCTCAAGCTGATGTAGAATGGAAGTTCTATGATGCACAGCCTTGTAGTGACAAAGCTTATAAAATAGAAGAATTATTCTATTCTTATGCCACACATTCTGACAAATTCACAGATGGTGTATGCCTATTTTGGAATTGCAATGTCGATAGATATCCTGCTAATTCCATTGTTTGTAGATTTGACACTAGAGTGCTATCTAACCTTAACTTGCCTGGTTGTGATGGTGGCAGTTTGTATGTAAATAAACATGCATTCCACACACCAGCTTTTGATAAAAGTGCTTTTGTTAATTTAAAACAATTACCATTTTTCTATTACTCTGACAGTCCATGTGAGTCTCATGGAAAACAAGTAGTGTCAGATATAGATTATGTACCACTAAAGTCTGCTACGTGTATAACACGTTGCAATTTAGGTGGTGCTGTCTGTAGACATCATGCTAATGAGTACAGATTGTATCTCGATGCTTATAACATGATGATCTCAGCTGGCTTTAGCTTGTGGGTTTACAAACAATTTGATACTTATAACCTCTGGAACACTTTTACAAGACTTCAGAGTTTAGAAAATGTGGCTTTTAATGTTGTAAATAAGGGACACTTTGATGGACAACAGGGTGAAGTACCAGTTTCTATCATTAATAACACTGTTTACACAAAAGTTGATGGTGTTGATGTAGAATTGTTTGAAAATAAAACAACATTACCTGTTAATGTAGCATTTGAGCTTTGGGCTAAGCGCAACATTAAACCAGTACCAGAGGTGAAAATACTCAATAATTTGGGTGTGGACATTGCTGCTAATACTGTGATCTGGGACTACAAAAGAGATGCTCCAGCACATATATCTACTATTGGTGTTTGTTCTATGACTGACATAGCCAAGAAACCAACTGAAACGATTTGTGCACCACTCACTGTCTTTTTTGATGGTAGAGTTGATGGTCAAGTAGACTTATTTAGAAATGCCCGTAATGGTGTTCTTATTACAGAAGGTAGTGTTAAAGGTTTACAACCATCTGTAGGTCCCAAACAAGCTAGTCTTAATGGAGTCACATTAATTGGAGAAGCCGTAAAAACACAGTTCAATTATTATAAGAAAGTTGATGGTGTTGTCCAACAATTACCTGAAACTTACTTTACTCAGAGTAGAAATTTACAAGAATTTAAACCCAGGAGTCAAATGGAAATTGATTTCTTAGAATTAGCTATGGATGAATTCATTGAACGGTATAAATTAGAAGGCTATGCCTTCGAACATATCGTTTATGGAGATTTTAGTCATAGTCAGTTAGGTGGTTTACATCTACTGATTGGACTAGCTAAACGTTTTAAGGAATCACCTTTTGAATTAGAAGATTTTATTCCTATGGACAGTACAGTTAAAAACTATTTCATAACAGATGCGCAAACAGGTTCATCTAAGTGTGTGTGTTCTGTTATTGATTTATTACTTGATGATTTTGTTGAAATAATAAAATCCCAAGATTTATCTGTAGTTTCTAAGGTTGTCAAAGTGACTATTGACTATACAGAAATTTCATTTATGCTTTGGTGTAAAGATGGCCATGTAGAAACATTTTACCCAAAATTACAATCTAGTCAAGCGTGGCAACCGGGTGTTGCTATGCCTAATCTTTACAAAATGCAAAGAATGCTATTAGAAAAGTGTGACCTTCAAAATTATGGTGATAGTGCAACATTACCTAAAGGCATAATGATGAATGTCGCAAAATATACTCAACTGTGTCAATATTTAAACACATTAACATTAGCTGTACCCTATAATATGAGAGTTATACATTTTGGTGCTGGTTCTGATAAAGGAGTTGCACCAGGTACAGCTGTTTTAAGACAGTGGTTGCCTACGGGTACGCTGCTTGTCGATTCAGATCTTAATGACTTTGTCTCTGATGCAGATTCAACTTTGATTGGTGATTGTGCAACTGTACATACAGCTAATAAATGGGATCTCATTATTAGTGATATGTACGACCCTAAGACTAAAAATGTTACAAAAGAAAATGACTCTAAAGAGGGTTTTTTCACTTACATTTGTGGGTTTATACAACAAAAGCTAGCTCTTGGAGGTTCCGTGGCTATAAAGATAACAGAACATTCTTGGAATGCTGATCTTTATAAGCTCATGGGACACTTCGCATGGTGGACAGCCTTTGTTACTAATGTGAATGCGTCATCATCTGAAGCATTTTTAATTGGATGTAATTATCTTGGCAAACCACGCGAACAAATAGATGGTTATGTCATGCATGCAAATTACATATTTTGGAGGAATACAAATCCAATTCAGTTGTCTTCCTATTCTTTATTTGACATGAGTAAATTTCCCCTTAAATTAAGGGGTACTGCTGTTATGTCTTTAAAAGAAGGTCAAATCAATGATATGATTTTATCTCTTCTTAGTAAAGGTAGACTTATAATTAGAGAAAACAACAGAGTTGTTATTTCTAGTGATGTTCTTGTTAACAACTAAACGAACAATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAACCAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCTGACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTTTTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGGTTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCATTGAGAAGTCTAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTACTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAATGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGAGTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTCTTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTAAGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCGTGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATTAACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTGATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCTAGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGTGCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAAGGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTCCTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCTGTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTATATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAATGATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCAGACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAGATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGGTGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAGAGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAAGGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGGTTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACTGTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACTTCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTTCCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGACACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCAGGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAGTCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGTTCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACTCATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGACTAATTCTCCGCGGCGGGCACGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATGTCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAAATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGTAGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAATATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAGACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAATTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAGCAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGCTTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTGCACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGCTCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCAGGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTGGAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATAGTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACTTCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAGCTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACAAAGTTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGACATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGCTGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGAAAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCATGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCATGATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGTTTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGTGTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTGCAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACATCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTCAAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCGATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATGGCCATGGTACATTTGGCTAGGTTTTATAGCTGGCTTGATTGCCATAGTAATGGTGACAATTATGCTTTGCTGTATGACCAGTTGCTGTAGTTGTCTCAAGGGCTGTTGTTCTTGTGGATCCTGCTGCAAATTTGATGAAGACGACTCTGAGCCAGTGCTCAAAGGAGTCAAATTACATTACACATAAACGAACTTATGGATTTGTTTATGAGAATCTTCACAATTGGAACTGTAACTTTGAAGCAAGGTGAAATCAAGGATGCTACTCCTTCAGATTTTGTTCGCGCTACTGCAACGATACCGATACAAGCCTCACTCCCTTTCGGATGGCTTATTGTTGGCGTTGCACTTCTTGCTGTTTTTCAGAGCGCTTCCAAAATCATAACCCTCAAAAAGAGATGGCAACTAGCACTCTCCAAGGGTGTTCACTTTGTTTGCAACTTGCTGTTGTTGTTTGTAACAGTTTACTCACACCTTTTGCTCGTTGCTGCTGGCCTTGAAGCCCCTTTTCTCTATCTTTATGCTTTAGTCTACTTCTTGCAGAGTATAAACTTTGTAAGAATAATAATGAGGCTTTGGCTTTGCTGGAAATGCCGTTCCAAAAACCCATTACTTTATGATGCCAACTATTTTCTTTGCTGGCATACTAATTGTTACGACTATTGTATACCTTACAATAGTGTAACTTCTTCAATTGTCATTACTTCAGGTGATGGCACAACAAGTCCTATTTCTGAACATGACTACCAGATTGGTGGTTATACTGAAAAATGGGAATCTGGAGTAAAAGACTGTGTTGTATTACACAGTTACTTCACTTCAGACTATTACCAGCTGTACTCAACTCAATTGAGTACAGACACTGGTGTTGAACATGTTACCTTCTTCATCTACAATAAAATTGTTGATGAGCCTGAAGAACATGTCCAAATTCACACAATCGACGGTTCATCCGGAGTTGTTAATCCAGTAATGGAACCAATTTATGATGAACCGACGACGACTACTAGCGTGCCTTTGTAAGCACAAGCTGATGAGTACGAACTTATGTACTCATTCGTTTCGGAAGAGACAGGTACGTTAATAGTTAATAGCGTACTTCTTTTTCTTGCTTTCGTGGTATTCTTGCTAGTTACACTAGCCATCCTTACTGCGCATCGATTGTGTGCGTACTGCTGCAATATTGTTAACGTGAGTCTTGTAAAACCTTCTTTTTACGTTTACTCTCGTGTTAAAAATCTGAATTCTTCTAGAGTTCCTGATCTTCTGGTCTAAACGAACTAAATATTATATTAGTTTTTCTGTTTGGAACTTTAATTTTAGCCATGGCAGATTCCAACGGTACTATTACCGTTGAAGAGCTTAAAAAGCTCCTTGAACAATGGAACCTAGTAATAGGTTTCCTATTCCTTACATGGATTTGTCTTCTACAATTTGCCTATGCCAACAGGAATAGGTTTTTGTATATAATTAAGTTAATTTTCCTCTGGCTGTTATGGCCAGTAACTTTAGCTTGTTTTGTGCTTGCTGCTGTTTACAGAATAAATTGGATCACCGGTGGAATTGCTATCGCAATGGCTTGTCTTGTAGGCTTGATGTGGCTCAGCTACTTCATTGCTTCTTTCAGACTGTTTGCGCGTACGCGTTCCATGTGGTCATTCAATCCAGAAACTAACATTCTTCTCAACGTGCCACTCCATGGCACTATTCTGACCAGACCGCTTCTAGAAAGTGAACTCGTAATCGGAGCTGTGATCCTTCGTGGACATCTTCGTATTGCTGGACACCATCTAGGACGCTGTGACATCAAGGACCTGCCTAAAGAAATCACTGTTGCTACATCACGAACGCTTTCTTATTACAAATTGGGAGCTTCGCAGCGTGTAGCAGGTGACTCAGGTTTTGCTGCATACAGTCGCTACAGGATTGGCAACTATAAATTAAACACAGACCATTCCAGTAGCAGTGACAATATTGCTTTGCTTGTACAGTAAGTGACAACAGATGTTTCATCTCGTTGACTTTCAGGTTACTATAGCAGAGATATTACTAATTATTATGAGGACTTTTAAAGTTTCCATTTGGAATCTTGATTACATCATAAACCTCATAATTAAAAATTTATCTAAGTCACTAACTGAGAATAAATATTCTCAATTAGATGAAGAGCAACCAATGGAGATTGATTAAACGAACATGAAAATTATTCTTTTCTTGGCACTGATAACACTCGCTACTTGTGAGCTTTATCACTACCAAGAGTGTGTTAGAGGTACAACAGTACTTTTAAAAGAACCTTGCTCTTCTGGAACATACGAGGGCAATTCACCATTTCATCCTCTAGCTGATAACAAATTTGCACTGACTTGCTTTAGCACTCAATTTGCTTTTGCTTGTCCTGACGGCGTAAAACACGTCTATCAGTTACGTGCCAGATCAGTTTCACCTAAACTGTTCATCAGACAAGAGGAAGTTCAAGAACTTTACTCTCCAATTTTTCTTATTGTTGCGGCAATAGTGTTTATAACACTTTGCTTCACACTCAAAAGAAAGACAGAATGATTGAACTTTCATTAATTGACTTCTATTTGTGCTTTTTAGCCTTTCTGCTATTCCTTGTTTTAATTATGCTTATTATCTTTTGGTTCTCACTTGAACTGCAAGATCATAATGAAACTTGTCACGCCTAAACGAACATGAAATTTCTTGTTTTCTTAGGAATCATCACAACTGTAGCTGCATTTCACCAAGAATGTAGTTTACAGTCATGTACTCAACATCAACCATATGTAGTTGATGACCCGTGTCCTATTCACTTCTATTCTAAATGGTATATTAGAGTAGGAGCTAGAAAATCAGCACCTTTAATTGAATTGTGCGTGGATGAGGCTGGTTCTAAATCACCCATTCAGTACATCGATATCGGTAATTATACAGTTTCCTGTTCACCTTTTACAATTAATTGCCAGGAACCTAAATTGGGTAGTCTTGTAGTGCGTTGTTCGTTCTATGAAGACTTTTTAGAGTATCATGACGTTCGTGTTGTTTTAGATTTCATCTAAACGAACAAACTAAAATGTCTGATAATGGACCCCAAAATCAGCGAAATGCACCCCGCATTACGTTTGGTGGACCCTCAGATTCAACTGGCAGTAACCAGAATGGAGAACGCAGTGGGGCGCGATCAAAACAACGTCGGCCCCAAGGTTTACCCAATAATACTGCGTCTTGGTTCACCGCTCTCACTCAACATGGCAAGGAAGACCTTAAATTCCCTCGAGGACAAGGCGTTCCAATTAACACCAATAGCAGTCCAGATGACCAAATTGGCTACTACCGAAGAGCTACCAGACGAATTCGTGGTGGTGACGGTAAAATGAAAGATCTCAGTCCAAGATGGTATTTCTACTACCTAGGAACTGGGCCAGAAGCTGGACTTCCCTATGGTGCTAACAAAGACGGCATCATATGGGTTGCAACTGAGGGAGCCTTGAATACACCAAAAGATCACATTGGCACCCGCAATCCTGCTAACAATGCTGCAATCGTGCTACAACTTCCTCAAGGAACAACATTGCCAAAAGGCTTCTACGCAGAAGGGAGCAGAGGCGGCAGTCAAGCCTCTTCTCGTTCCTCATCACGTAGTCGCAACAGTTCAAGAAATTCAACTCCAGGCAGCAGTAGGGGAACTTCTCCTGCTAGAATGGCTGGCAATGGCGGTGATGCTGCTCTTGCTTTGCTGCTGCTTGACAGATTGAACCAGCTTGAGAGCAAAATGTCTGGTAAAGGCCAACAACAACAAGGCCAAACTGTCACTAAGAAATCTGCTGCTGAGGCTTCTAAGAAGCCTCGGCAAAAACGTACTGCCACTAAAGCATACAATGTAACACAAGCTTTCGGCAGACGTGGTCCAGAACAAACCCAAGGAAATTTTGGGGACCAGGAACTAATCAGACAAGGAACTGATTACAAACATTGGCCGCAAATTGCACAATTTGCCCCCAGCGCTTCAGCGTTCTTCGGAATGTCGCGCATTGGCATGGAAGTCACACCTTCGGGAACGTGGTTGACCTACACAGGTGCCATCAAATTGGATGACAAAGATCCAAATTTCAAAGATCAAGTCATTTTGCTGAATAAGCATATTGACGCATACAAAACATTCCCATCAACAGAGCCTAAAAAGGACAAAAAGAAGAAGGCTGATGAAACTCAAGCCTTACCGCAGAGACAGAAGAAACAGCAAACTGTGACTCTTCTTCCTGCTGCAGATTTGGATGATTTCTCCAAACAATTGCAACAATCCATGAGCAGTGCTGACTCAACTCAGGCCTAAACTCATGCAGACCACACAAGGCAGATGGGCTATATAAACGTTTTCGCTTTTCCGTTTACGATGTATAGTCTACTCTTGTGCAGAATGAATTCTCGTAACTACATAGCACAAGTAGATGTAGTTAACTTTAATCTCACATAGCAATCTTTAATCAGTGTGTAACATTAGGGAGGACTTGAAAGAGCCACCACATTTTCACCGAGGCCACGCGGAGTACGATCGAGTGTACAGTGAACAATGCTAGGGAGAGCTGCCTATATGGAAGAGCCCTAATGTGTAAAATTAATTTTAGTAGTGCTATCCNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNNN MN908947.3 ATTAAAGGTTTATACCTTCCCAGGTAACAAACCAACCAACTTTCGATCTCTTGTAGATCTGTTCTCTAAACGAACTTTAAAATCTGTGTGGCTGTCACTCGGCTGCATGCTTAGTGCACTCACGCAGTATAATTAATAACTAATTACTGTCGTTGACAGGACACGAGTAACTCGTCTATCTTCTGCAGGCTGCTTACGGTTTCGTCCGTGTTGCAGCCGATCATCAGCACATCTAGGTTTCGTCCGGGTGTGACCGAAAGGTAAGATGGAGAGCCTTGTCCCTGGTTTCAACGAGAAAACACACGTCCAACTCAGTTTGCCTGTTTTACAGGTTCGCGACGTGCTCGTACGTGGCTTTGGAGACTCCGTGGAGGAGGTCTTATCAGAGGCACGTCAACATCTTAAAGATGGCACTTGTGGCTTAGTAGAAGTTGAAAAAGGCGTTTTGCCTCAACTTGAACAGCCCTATGTGTTCATCAAACGTTCGGATGCTCGAACTGCACCTCATGGTCATGTTATGGTTGAGCTGGTAGCAGAACTCGAAGGCATTCAGTACGGTCGTAGTGGTGAGACACTTGGTGTCCTTGTCCCTCATGTGGGCGAAATACCAGTGGCTTACCGCAAGGTTCTTCTTCGTAAGAACGGTAATAAAGGAGCTGGTGGCCATAGTTACGGCGCCGATCTAAAGTCATTTGACTTAGGCGACGAGCTTGGCACTGATCCTTATGAAGATTTTCAAGAAAACTGGAACACTAAACATAGCAGTGGTGTTACCCGTGAACTCATGCGTGAGCTTAACGGAGGGGCATACACTCGCTATGTCGATAACAACTTCTGTGGCCCTGATGGCTACCCTCTTGAGTGCATTAAAGACCTTCTAGCACGTGCTGGTAAAGCTTCATGCACTTTGTCCGAACAACTGGACTTTATTGACACTAAGAGGGGTGTATACTGCTGCCGTGAACATGAGCATGAAATTGCTTGGTACACGGAACGTTCTGAAAAGAGCTATGAATTGCAGACACCTTTTGAAATTAAATTGGCAAAGAAATTTGACACCTTCAATGGGGAATGTCCAAATTTTGTATTTCCCTTAAATTCCATAATCAAGACTATTCAACCAAGGGTTGAAAAGAAAAAGCTTGATGGCTTTATGGGTAGAATTCGATCTGTCTATCCAGTTGCGTCACCAAATGAATGCAACCAAATGTGCCTTTCAACTCTCATGAAGTGTGATCATTGTGGTGAAACTTCATGGCAGACGGGCGATTTTGTTAAAGCCACTTGCGAATTTTGTGGCACTGAGAATTTGACTAAAGAAGGTGCCACTACTTGTGGTTACTTACCCCAAAATGCTGTTGTTAAAATTTATTGTCCAGCATGTCACAATTCAGAAGTAGGACCTGAGCATAGTCTTGCCGAATACCATAATGAATCTGGCTTGAAAACCATTCTTCGTAAGGGTGGTCGCACTATTGCCTTTGGAGGCTGTGTGTTCTCTTATGTTGGTTGCCATAACAAGTGTGCCTATTGGGTTCCACGTGCTAGCGCTAACATAGGTTGTAACCATACAGGTGTTGTTGGAGAAGGTTCCGAAGGTCTTAATGACAACCTTCTTGAAATACTCCAAAAAGAGAAAGTCAACATCAATATTGTTGGTGACTTTAAACTTAATGAAGAGATCGCCATTATTTTGGCATCTTTTTCTGCTTCCACAAGTGCTTTTGTGGAAACTGTGAAAGGTTTGGATTATAAAGCATTCAAACAAATTGTTGAATCCTGTGGTAATTTTAAAGTTACAAAAGGAAAAGCTAAAAAAGGTGCCTGGAATATTGGTGAACAGAAATCAATACTGAGTCCTCTTTATGCATTTGCATCAGAGGCTGCTCGTGTTGTACGATCAATTTTCTCCCGCACTCTTGAAACTGCTCAAAATTCTGTGCGTGTTTTACAGAAGGCCGCTATAACAATACTAGATGGAATTTCACAGTATTCACTGAGACTCATTGATGCTATGATGTTCACATCTGATTTGGCTACTAACAATCTAGTTGTAATGGCCTACATTACAGGTGGTGTTGTTCAGTTGACTTCGCAGTGGCTAACTAACATCTTTGGCACTGTTTATGAAAAACTCAAACCCGTCCTTGATTGGCTTGAAGAGAAGTTTAAGGAAGGTGTAGAGTTTCTTAGAGACGGTTGGGAAATTGTTAAATTTATCTCAACCTGTGCTTGTGAAATTGTCGGTGGACAAATTGTCACCTGTGCAAAGGAAATTAAGGAGAGTGTTCAGACATTCTTTAAGCTTGTAAATAAATTTTTGGCTTTGTGTGCTGACTCTATCATTATTGGTGGAGCTAAACTTAAAGCCTTGAATTTAGGTGAAACATTTGTCACGCACTCAAAGGGATTGTACAGAAAGTGTGTTAAATCCAGAGAAGAAACTGGCCTACTCATGCCTCTAAAAGCCCCAAAAGAAATTATCTTCTTAGAGGGAGAAACACTTCCCACAGAAGTGTTAACAGAGGAAGTTGTCTTGAAAACTGGTGATTTACAACCATTAGAACAACCTACTAGTGAAGCTGTTGAAGCTCCATTGGTTGGTACACCAGTTTGTATTAACGGGCTTATGTTGCTCGAAATCAAAGACACAGAAAAGTACTGTGCCCTTGCACCTAATATGATGGTAACAAACAATACCTTCACACTCAAAGGCGGTGCACCAACAAAGGTTACTTTTGGTGATGACACTGTGATAGAAGTGCAAGGTTACAAGAGTGTGAATATCACTTTTGAACTTGATGAAAGGATTGATAAAGTACTTAATGAGAAGTGCTCTGCCTATACAGTTGAACTCGGTACAGAAGTAAATGAGTTCGCCTGTGTTGTGGCAGATGCTGTCATAAAAACTTTGCAACCAGTATCTGAATTACTTACACCACTGGGCATTGATTTAGATGAGTGGAGTATGGCTACATACTACTTATTTGATGAGTCTGGTGAGTTTAAATTGGCTTCACATATGTATTGTTCTTTCTACCCTCCAGATGAGGATGAAGAAGAAGGTGATTGTGAAGAAGAAGAGTTTGAGCCATCAACTCAATATGAGTATGGTACTGAAGATGATTACCAAGGTAAACCTTTGGAATTTGGTGCCACTTCTGCTGCTCTTCAACCTGAAGAAGAGCAAGAAGAAGATTGGTTAGATGATGATAGTCAACAAACTGTTGGTCAACAAGACGGCAGTGAGGACAATCAGACAACTACTATTCAAACAATTGTTGAGGTTCAACCTCAATTAGAGATGGAACTTACACCAGTTGTTCAGACTATTGAAGTGAATAGTTTTAGTGGTTATTTAAAACTTACTGACAATGTATACATTAAAAATGCAGACATTGTGGAAGAAGCTAAAAAGGTAAAACCAACAGTGGTTGTTAATGCAGCCAATGTTTACCTTAAACATGGAGGAGGTGTTGCAGGAGCCTTAAATAAGGCTACTAACAATGCCATGCAAGTTGAATCTGATGATTACATAGCTACTAATGGACCACTTAAAGTGGGTGGTAGTTGTGTTTTAAGCGGACACAATCTTGCTAAACACTGTCTTCATGTTGTCGGCCCAAATGTTAACAAAGGTGAAGACATTCAACTTCTTAAGAGTGCTTATGAAAATTTTAATCAGCACGAAGTTCTACTTGCACCATTATTATCAGCTGGTATTTTTGGTGCTGACCCTATACATTCTTTAAGAGTTTGTGTAGATACTGTTCGCACAAATGTCTACTTAGCTGTCTTTGATAAAAATCTCTATGACAAACTTGTTTCAAGCTTTTTGGAAATGAAGAGTGAAAAGCAAGTTGAACAAAAGATCGCTGAGATTCCTAAAGAGGAAGTTAAGCCATTTATAACTGAAAGTAAACCTTCAGTTGAACAGAGAAAACAAGATGATAAGAAAATCAAAGCTTGTGTTGAAGAAGTTACAACAACTCTGGAAGAAACTAAGTTCCTCACAGAAAACTTGTTACTTTATATTGACATTAATGGCAATCTTCATCCAGATTCTGCCACTCTTGTTAGTGACATTGACATCACTTTCTTAAAGAAAGATGCTCCATATATAGTGGGTGATGTTGTTCAAGAGGGTGTTTTAACTGCTGTGGTTATACCTACTAAAAAGGCTGGTGGCACTACTGAAATGCTAGCGAAAGCTTTGAGAAAAGTGCCAACAGACAATTATATAACCACTTACCCGGGTCAGGGTTTAAATGGTTACACTGTAGAGGAGGCAAAGACAGTGCTTAAAAAGTGTAAAAGTGCCTTTTACATTCTACCATCTATTATCTCTAATGAGAAGCAAGAAATTCTTGGAACTGTTTCTTGGAATTTGCGAGAAATGCTTGCACATGCAGAAGAAACACGCAAATTAATGCCTGTCTGTGTGGAAACTAAAGCCATAGTTTCAACTATACAGCGTAAATATAAGGGTATTAAAATACAAGAGGGTGTGGTTGATTATGGTGCTAGATTTTACTTTTACACCAGTAAAACAACTGTAGCGTCACTTATCAACACACTTAACGATCTAAATGAAACTCTTGTTACAATGCCACTTGGCTATGTAACACATGGCTTAAATTTGGAAGAAGCTGCTCGGTATATGAGATCTCTCAAAGTGCCAGCTACAGTTTCTGTTTCTTCACCTGATGCTGTTACAGCGTATAATGGTTATCTTACTTCTTCTTCTAAAACACCTGAAGAACATTTTATTGAAACCATCTCACTTGCTGGTTCCTATAAAGATTGGTCCTATTCTGGACAATCTACACAACTAGGTATAGAATTTCTTAAGAGAGGTGATAAAAGTGTATATTACACTAGTAATCCTACCACATTCCACCTAGATGGTGAAGTTATCACCTTTGACAATCTTAAGACACTTCTTTCTTTGAGAGAAGTGAGGACTATTAAGGTGTTTACAACAGTAGACAACATTAACCTCCACACGCAAGTTGTGGACATGTCAATGACATATGGACAACAGTTTGGTCCAACTTATTTGGATGGAGCTGATGTTACTAAAATAAAACCTCATAATTCACATGAAGGTAAAACATTTTATGTTTTACCTAATGATGACACTCTACGTGTTGAGGCTTTTGAGTACTACCACACAACTGATCCTAGTTTTCTGGGTAGGTACATGTCAGCATTAAATCACACTAAAAAGTGGAAATACCCACAAGTTAATGGTTTAACTTCTATTAAATGGGCAGATAACAACTGTTATCTTGCCACTGCATTGTTAACACTCCAACAAATAGAGTTGAAGTTTAATCCACCTGCTCTACAAGATGCTTATTACAGAGCAAGGGCTGGTGAAGCTGCTAACTTTTGTGCACTTATCTTAGCCTACTGTAATAAGACAGTAGGTGAGTTAGGTGATGTTAGAGAAACAATGAGTTACTTGTTTCAACATGCCAATTTAGATTCTTGCAAAAGAGTCTTGAACGTGGTGTGTAAAACTTGTGGACAACAGCAGACAACCCTTAAGGGTGTAGAAGCTGTTATGTACATGGGCACACTTTCTTATGAACAATTTAAGAAAGGTGTTCAGATACCTTGTACGTGTGGTAAACAAGCTACAAAATATCTAGTACAACAGGAGTCACCTTTTGTTATGATGTCAGCACCACCTGCTCAGTATGAACTTAAGCATGGTACATTTACTTGTGCTAGTGAGTACACTGGTAATTACCAGTGTGGTCACTATAAACATATAACTTCTAAAGAAACTTTGTATTGCATAGACGGTGCTTTACTTACAAAGTCCTCAGAATACAAAGGTCCTATTACGGATGTTTTCTACAAAGAAAACAGTTACACAACAACCATAAAACCAGTTACTTATAAATTGGATGGTGTTGTTTGTACAGAAATTGACCCTAAGTTGGACAATTATTATAAGAAAGACAATTCTTATTTCACAGAGCAACCAATTGATCTTGTACCAAACCAACCATATCCAAACGCAAGCTTCGATAATTTTAAGTTTGTATGTGATAATATCAAATTTGCTGATGATTTAAACCAGTTAACTGGTTATAAGAAACCTGCTTCAAGAGAGCTTAAAGTTACATTTTTCCCTGACTTAAATGGTGATGTGGTGGCTATTGATTATAAACACTACACACCCTCTTTTAAGAAAGGAGCTAAATTGTTACATAAACCTATTGTTTGGCATGTTAACAATGCAACTAATAAAGCCACGTATAAACCAAATACCTGGTGTATACGTTGTCTTTGGAGCACAAAACCAGTTGAAACATCAAATTCGTTTGATGTACTGAAGTCAGAGGACGCGCAGGGAATGGATAATCTTGCCTGCGAAGATCTAAAACCAGTCTCTGAAGAAGTAGTGGAAAATCCTACCATACAGAAAGACGTTCTTGAGTGTAATGTGAAAACTACCGAAGTTGTAGGAGACATTATACTTAAACCAGCAAATAATAGTTTAAAAATTACAGAAGAGGTTGGCCACACAGATCTAATGGCTGCTTATGTAGACAATTCTAGTCTTACTATTAAGAAACCTAATGAATTATCTAGAGTATTAGGTTTGAAAACCCTTGCTACTCATGGTTTAGCTGCTGTTAATAGTGTCCCTTGGGATACTATAGCTAATTATGCTAAGCCTTTTCTTAACAAAGTTGTTAGTACAACTACTAACATAGTTACACGGTGTTTAAACCGTGTTTGTACTAATTATATGCCTTATTTCTTTACTTTATTGCTACAATTGTGTACTTTTACTAGAAGTACAAATTCTAGAATTAAAGCATCTATGCCGACTACTATAGCAAAGAATACTGTTAAGAGTGTCGGTAAATTTTGTCTAGAGGCTTCATTTAATTATTTGAAGTCACCTAATTTTTCTAAACTGATAAATATTATAATTTGGTTTTTACTATTAAGTGTTTGCCTAGGTTCTTTAATCTACTCAACCGCTGCTTTAGGTGTTTTAATGTCTAATTTAGGCATGCCTTCTTACTGTACTGGTTACAGAGAAGGCTATTTGAACTCTACTAATGTCACTATTGCAACCTACTGTACTGGTTCTATACCTTGTAGTGTTTGTCTTAGTGGTTTAGATTCTTTAGACACCTATCCTTCTTTAGAAACTATACAAATTACCATTTCATCTTTTAAATGGGATTTAACTGCTTTTGGCTTAGTTGCAGAGTGGTTTTTGGCATATATTCTTTTCACTAGGTTTTTCTATGTACTTGGATTGGCTGCAATCATGCAATTGTTTTTCAGCTATTTTGCAGTACATTTTATTAGTAATTCTTGGCTTATGTGGTTAATAATTAATCTTGTACAAATGGCCCCGATTTCAGCTATGGTTAGAATGTACATCTTCTTTGCATCATTTTATTATGTATGGAAAAGTTATGTGCATGTTGTAGACGGTTGTAATTCATCAACTTGTATGATGTGTTACAAACGTAATAGAGCAACAAGAGTCGAATGTACAACTATTGTTAATGGTGTTAGAAGGTCCTTTTATGTCTATGCTAATGGAGGTAAAGGCTTTTGCAAACTACACAATTGGAATTGTGTTAATTGTGATACATTCTGTGCTGGTAGTACATTTATTAGTGATGAAGTTGCGAGAGACTTGTCACTACAGTTTAAAAGACCAATAAATCCTACTGACCAGTCTTCTTACATCGTTGATAGTGTTACAGTGAAGAATGGTTCCATCCATCTTTACTTTGATAAAGCTGGTCAAAAGACTTATGAAAGACATTCTCTCTCTCATTTTGTTAACTTAGACAACCTGAGAGCTAATAACACTAAAGGTTCATTGCCTATTAATGTTATAGTTTTTGATGGTAAATCAAAATGTGAAGAATCATCTGCAAAATCAGCGTCTGTTTACTACAGTCAGCTTATGTGTCAACCTATACTGTTACTAGATCAGGCATTAGTGTCTGATGTTGGTGATAGTGCGGAAGTTGCAGTTAAAATGTTTGATGCTTACGTTAATACGTTTTCATCAACTTTTAACGTACCAATGGAAAAACTCAAAACACTAGTTGCAACTGCAGAAGCTGAACTTGCAAAGAATGTGTCCTTAGACAATGTCTTATCTACTTTTATTTCAGCAGCTCGGCAAGGGTTTGTTGATTCAGATGTAGAAACTAAAGATGTTGTTGAATGTCTTAAATTGTCACATCAATCTGACATAGAAGTTACTGGCGATAGTTGTAATAACTATATGCTCACCTATAACAAAGTTGAAAACATGACACCCCGTGACCTTGGTGCTTGTATTGACTGTAGTGCGCGTCATATTAATGCGCAGGTAGCAAAAAGTCACAACATTGCTTTGATATGGAACGTTAAAGATTTCATGTCATTGTCTGAACAACTACGAAAACAAATACGTAGTGCTGCTAAAAAGAATAACTTACCTTTTAAGTTGACATGTGCAACTACTAGACAAGTTGTTAATGTTGTAACAACAAAGATAGCACTTAAGGGTGGTAAAATTGTTAATAATTGGTTGAAGCAGTTAATTAAAGTTACACTTGTGTTCCTTTTTGTTGCTGCTATTTTCTATTTAATAACACCTGTTCATGTCATGTCTAAACATACTGACTTTTCAAGTGAAATCATAGGATACAAGGCTATTGATGGTGGTGTCACTCGTGACATAGCATCTACAGATACTTGTTTTGCTAACAAACATGCTGATTTTGACACATGGTTTAGCCAGCGTGGTGGTAGTTATACTAATGACAAAGCTTGCCCATTGATTGCTGCAGTCATAACAAGAGAAGTGGGTTTTGTCGTGCCTGGTTTGCCTGGCACGATATTACGCACAACTAATGGTGACTTTTTGCATTTCTTACCTAGAGTTTTTAGTGCAGTTGGTAACATCTGTTACACACCATCAAAACTTATAGAGTACACTGACTTTGCAACATCAGCTTGTGTTTTGGCTGCTGAATGTACAATTTTTAAAGATGCTTCTGGTAAGCCAGTACCATATTGTTATGATACCAATGTACTAGAAGGTTCTGTTGCTTATGAAAGTTTACGCCCTGACACACGTTATGTGCTCATGGATGGCTCTATTATTCAATTTCCTAACACCTACCTTGAAGGTTCTGTTAGAGTGGTAACAACTTTTGATTCTGAGTACTGTAGGCACGGCACTTGTGAAAGATCAGAAGCTGGTGTTTGTGTATCTACTAGTGGTAGATGGGTACTTAACAATGATTATTACAGATCTTTACCAGGAGTTTTCTGTGGTGTAGATGCTGTAAATTTACTTACTAATATGTTTACACCACTAATTCAACCTATTGGTGCTTTGGACATATCAGCATCTATAGTAGCTGGTGGTATTGTAGCTATCGTAGTAACATGCCTTGCCTACTATTTTATGAGGTTTAGAAGAGCTTTTGGTGAATACAGTCATGTAGTTGCCTTTAATACTTTACTATTCCTTATGTCATTCACTGTACTCTGTTTAACACCAGTTTACTCATTCTTACCTGGTGTTTATTCTGTTATTTACTTGTACTTGACATTTTATCTTACTAATGATGTTTCTTTTTTAGCACATATTCAGTGGATGGTTATGTTCACACCTTTAGTACCTTTCTGGATAACAATTGCTTATATCATTTGTATTTCCACAAAGCATTTCTATTGGTTCTTTAGTAATTACCTAAAGAGACGTGTAGTCTTTAATGGTGTTTCCTTTAGTACTTTTGAAGAAGCTGCGCTGTGCACCTTTTTGTTAAATAAAGAAATGTATCTAAAGTTGCGTAGTGATGTGCTATTACCTCTTACGCAATATAATAGATACTTAGCTCTTTATAATAAGTACAAGTATTTTAGTGGAGCAATGGATACAACTAGCTACAGAGAAGCTGCTTGTTGTCATCTCGCAAAGGCTCTCAATGACTTCAGTAACTCAGGTTCTGATGTTCTTTACCAACCACCACAAACCTCTATCACCTCAGCTGTTTTGCAGAGTGGTTTTAGAAAAATGGCATTCCCATCTGGTAAAGTTGAGGGTTGTATGGTACAAGTAACTTGTGGTACAACTACACTTAACGGTCTTTGGCTTGATGACGTAGTTTACTGTCCAAGACATGTGATCTGCACCTCTGAAGACATGCTTAACCCTAATTATGAAGATTTACTCATTCGTAAGTCTAATCATAATTTCTTGGTACAGGCTGGTAATGTTCAACTCAGGGTTATTGGACATTCTATGCAAAATTGTGTACTTAAGCTTAAGGTTGATACAGCCAATCCTAAGACACCTAAGTATAAGTTTGTTCGCATTCAACCAGGACAGACTTTTTCAGTGTTAGCTTGTTACAATGGTTCACCATCTGGTGTTTACCAATGTGCTATGAGGCCCAATTTCACTATTAAGGGTTCATTCCTTAATGGTTCATGTGGTAGTGTTGGTTTTAACATAGATTATGACTGTGTCTCTTTTTGTTACATGCACCATATGGAATTACCAACTGGAGTTCATGCTGGCACAGACTTAGAAGGTAACTTTTATGGACCTTTTGTTGACAGGCAAACAGCACAAGCAGCTGGTACGGACACAACTATTACAGTTAATGTTTTAGCTTGGTTGTACGCTGCTGTTATAAATGGAGACAGGTGGTTTCTCAATCGATTTACCACAACTCTTAATGACTTTAACCTTGTGGCTATGAAGTACAATTATGAACCTCTAACACAAGACCATGTTGACATACTAGGACCTCTTTCTGCTCAAACTGGAATTGCCGTTTTAGATATGTGTGCTTCATTAAAAGAATTACTGCAAAATGGTATGAATGGACGTACCATATTGGGTAGTGCTTTATTAGAAGATGAATTTACACCTTTTGATGTTGTTAGACAATGCTCAGGTGTTACTTTCCAAAGTGCAGTGAAAAGAACAATCAAGGGTACACACCACTGGTTGTTACTCACAATTTTGACTTCACTTTTAGTTTTAGTCCAGAGTACTCAATGGTCTTTGTTCTTTTTTTTGTATGAAAATGCCTTTTTACCTTTTGCTATGGGTATTATTGCTATGTCTGCTTTTGCAATGATGTTTGTCAAACATAAGCATGCATTTCTCTGTTTGTTTTTGTTACCTTCTCTTGCCACTGTAGCTTATTTTAATATGGTCTATATGCCTGCTAGTTGGGTGATGCGTATTATGACATGGTTGGATATGGTTGATACTAGTTTGTCTGGTTTTAAGCTAAAAGACTGTGTTATGTATGCATCAGCTGTAGTGTTACTAATCCTTATGACAGCAAGAACTGTGTATGATGATGGTGCTAGGAGAGTGTGGACACTTATGAATGTCTTGACACTCGTTTATAAAGTTTATTATGGTAATGCTTTAGATCAAGCCATTTCCATGTGGGCTCTTATAATCTCTGTTACTTCTAACTACTCAGGTGTAGTTACAACTGTCATGTTTTTGGCCAGAGGTATTGTTTTTATGTGTGTTGAGTATTGCCCTATTTTCTTCATAACTGGTAATACACTTCAGTGTATAATGCTAGTTTATTGTTTCTTAGGCTATTTTTGTACTTGTTACTTTGGCCTCTTTTGTTTACTCAACCGCTACTTTAGACTGACTCTTGGTGTTTATGATTACTTAGTTTCTACACAGGAGTTTAGATATATGAATTCACAGGGACTACTCCCACCCAAGAATAGCATAGATGCCTTCAAACTCAACATTAAATTGTTGGGTGTTGGTGGCAAACCTTGTATCAAAGTAGCCACTGTACAGTCTAAAATGTCAGATGTAAAGTGCACATCAGTAGTCTTACTCTCAGTTTTGCAACAACTCAGAGTAGAATCATCATCTAAATTGTGGGCTCAATGTGTCCAGTTACACAATGACATTCTCTTAGCTAAAGATACTACTGAAGCCTTTGAAAAAATGGTTTCACTACTTTCTGTTTTGCTTTCCATGCAGGGTGCTGTAGACATAAACAAGCTTTGTGAAGAAATGCTGGACAACAGGGCAACCTTACAAGCTATAGCCTCAGAGTTTAGTTCCCTTCCATCATATGCAGCTTTTGCTACTGCTCAAGAAGCTTATGAGCAGGCTGTTGCTAATGGTGATTCTGAAGTTGTTCTTAAAAAGTTGAAGAAGTCTTTGAATGTGGCTAAATCTGAATTTGACCGTGATGCAGCCATGCAACGTAAGTTGGAAAAGATGGCTGATCAAGCTATGACCCAAATGTATAAACAGGCTAGATCTGAGGACAAGAGGGCAAAAGTTACTAGTGCTATGCAGACAATGCTTTTCACTATGCTTAGAAAGTTGGATAATGATGCACTCAACAACATTATCAACAATGCAAGAGATGGTTGTGTTCCCTTGAACATAATACCTCTTACAACAGCAGCCAAACTAATGGTTGTCATACCAGACTATAACACATATAAAAATACGTGTGATGGTACAACATTTACTTATGCATCAGCATTGTGGGAAATCCAACAGGTTGTAGATGCAGATAGTAAAATTGTTCAACTTAGTGAAATTAGTATGGACAATTCACCTAATTTAGCATGGCCTCTTATTGTAACAGCTTTAAGGGCCAATTCTGCTGTCAAATTACAGAATAATGAGCTTAGTCCTGTTGCACTACGACAGATGTCTTGTGCTGCCGGTACTACACAAACTGCTTGCACTGATGACAATGCGTTAGCTTACTACAACACAACAAAGGGAGGTAGGTTTGTACTTGCACTGTTATCCGATTTACAGGATTTGAAATGGGCTAGATTCCCTAAGAGTGATGGAACTGGTACTATCTATACAGAACTGGAACCACCTTGTAGGTTTGTTACAGACACACCTAAAGGTCCTAAAGTGAAGTATTTATACTTTATTAAAGGATTAAACAACCTAAATAGAGGTATGGTACTTGGTAGTTTAGCTGCCACAGTACGTCTACAAGCTGGTAATGCAACAGAAGTGCCTGCCAATTCAACTGTATTATCTTTCTGTGCTTTTGCTGTAGATGCTGCTAAAGCTTACAAAGATTATCTAGCTAGTGGGGGACAACCAATCACTAATTGTGTTAAGATGTTGTGTACACACACTGGTACTGGTCAGGCAATAACAGTTACACCGGAAGCCAATATGGATCAAGAATCCTTTGGTGGTGCATCGTGTTGTCTGTACTGCCGTTGCCACATAGATCATCCAAATCCTAAAGGATTTTGTGACTTAAAAGGTAAGTATGTACAAATACCTACAACTTGTGCTAATGACCCTGTGGGTTTTACACTTAAAAACACAGTCTGTACCGTCTGCGGTATGTGGAAAGGTTATGGCTGTAGTTGTGATCAACTCCGCGAACCCATGCTTCAGTCAGCTGATGCACAATCGTTTTTAAACGGGTTTGCGGTGTAAGTGCAGCCCGTCTTACACCGTGCGGCACAGGCACTAGTACTGATGTCGTATACAGGGCTTTTGACATCTACAATGATAAAGTAGCTGGTTTTGCTAAATTCCTAAAAACTAATTGTTGTCGCTTCCAAGAAAAGGACGAAGATGACAATTTAATTGATTCTTACTTTGTAGTTAAGAGACACACTTTCTCTAACTACCAACATGAAGAAACAATTTATAATTTACTTAAGGATTGTCCAGCTGTTGCTAAACATGACTTCTTTAAGTTTAGAATAGACGGTGACATGGTACCACATATATCACGTCAACGTCTTACTAAATACACAATGGCAGACCTCGTCTATGCTTTAAGGCATTTTGATGAAGGTAATTGTGACACATTAAAAGAAATACTTGTCACATACAATTGTTGTGATGATGATTATTTCAATAAAAAGGACTGGTATGATTTTGTAGAAAACCCAGATATATTACGCGTATACGCCAACTTAGGTGAACGTGTACGCCAAGCTTTGTTAAAAACAGTACAATTCTGTGATGCCATGCGAAATGCTGGTATTGTTGGTGTACTGACATTAGATAATCAAGATCTCAATGGTAACTGGTATGATTTCGGTGATTTCATACAAACCACGCCAGGTAGTGGAGTTCCTGTTGTAGATTCTTATTATTCATTGTTAATGCCTATATTAACCTTGACCAGGGCTTTAACTGCAGAGTCACATGTTGACACTGACTTAACAAAGCCTTACATTAAGTGGGATTTGTTAAAATATGACTTCACGGAAGAGAGGTTAAAACTCTTTGACCGTTATTTTAAATATTGGGATCAGACATACCACCCAAATTGTGTTAACTGTTTGGATGACAGATGCATTCTGCATTGTGCAAACTTTAATGTTTTATTCTCTACAGTGTTCCCACCTACAAGTTTTGGACCACTAGTGAGAAAAATATTTGTTGATGGTGTTCCATTTGTAGTTTCAACTGGATACCACTTCAGAGAGCTAGGTGTTGTACATAATCAGGATGTAAACTTACATAGCTCTAGACTTAGTTTTAAGGAATTACTTGTGTATGCTGCTGACCCTGCTATGCACGCTGCTTCTGGTAATCTATTACTAGATAAACGCACTACGTGCTTTTCAGTAGCTGCACTTACTAACAATGTTGCTTTTCAAACTGTCAAACCCGGTAATTTTAACAAAGACTTCTATGACTTTGCTGTGTCTAAGGGTTTCTTTAAGGAAGGAAGTTCTGTTGAATTAAAACACTTCTTCTTTGCTCAGGATGGTAATGCTGCTATCAGCGATTATGACTACTATCGTTATAATCTACCAACAATGTGTGATATCAGACAACTACTATTTGTAGTTGAAGTTGTTGATAAGTACTTTGATTGTTACGATGGTGGCTGTATTAATGCTAACCAAGTCATCGTCAACAACCTAGACAAATCAGCTGGTTTTCCATTTAATAAATGGGGTAAGGCTAGACTTTATTATGATTCAATGAGTTATGAGGATCAAGATGCACTTTTCGCATATACAAAACGTAATGTCATCCCTACTATAACTCAAATGAATCTTAAGTATGCCATTAGTGCAAAGAATAGAGCTCGCACCGTAGCTGGTGTCTCTATCTGTAGTACTATGACCAATAGACAGTTTCATCAAAAATTATTGAAATCAATAGCCGCCACTAGAGGAGCTACTGTAGTAATTGGAACAAGCAAATTCTATGGTGGTTGGCACAACATGTTAAAAACTGTTTATAGTGATGTAGAAAACCCTCACCTTATGGGTTGGGATTATCCTAAATGTGATAGAGCCATGCCTAACATGCTTAGAATTATGGCCTCACTTGTTCTTGCTCGCAAACATACAACGTGTTGTAGCTTGTCACACCGTTTCTATAGATTAGCTAATGAGTGTGCTCAAGTATTGAGTGAAATGGTCATGTGTGGCGGTTCACTATATGTTAAACCAGGTGGAACCTCATCAGGAGATGCCACAACTGCTTATGCTAATAGTGTTTTTAACATTTGTCAAGCTGTCACGGCCAATGTTAATGCACTTTTATCTACTGATGGTAACAAAATTGCCGATAAGTATGTCCGCAATTTACAACACAGACTTTATGAGTGTCTCTATAGAAATAGAGATGTTGACACAGACTTTGTGAATGAGTTTTACGCATATTTGCGTAAACATTTCTCAATGATGATACTCTCTGACGATGCTGTTGTGTGTTTCAATAGCACTTATGCATCTCAAGGTCTAGTGGCTAGCATAAAGAACTTTAAGTCAGTTCTTTATTATCAAAACAATGTTTTTATGTCTGAAGCAAAATGTTGGACTGAGACTGACCTTACTAAAGGACCTCATGAATTTTGCTCTCAACATACAATGCTAGTTAAACAGGGTGATGATTATGTGTACCTTCCTTACCCAGATCCATCAAGAATCCTAGGGGCCGGCTGTTTTGTAGATGATATCGTAAAAACAGATGGTACACTTATGATTGAACGGTTCGTGTCTTTAGCTATAGATGCTTACCCACTTACTAAACATCCTAATCAGGAGTATGCTGATGTCTTTCATTTGTACTTACAATACATAAGAAAGCTACATGATGAGTTAACAGGACACATGTTAGACATGTATTCTGTTATGCTTACTAATGATAACACTTCAAGGTATTGGGAACCTGAGTTTTATGAGGCTATGTACACACCGCATACAGTCTTACAGGCTGTTGGGGCTTGTGTTCTTTGCAATTCACAGACTTCATTAAGATGTGGTGCTTGCATACGTAGACCATTCTTATGTTGTAAATGCTGTTACGACCATGTCATATCAACATCACATAAATTAGTCTTGTCTGTTAATCCGTATGTTTGCAATGCTCCAGGTTGTGATGTCACAGATGTGACTCAACTTTACTTAGGAGGTATGAGCTATTATTGTAAATCACATAAACCACCCATTAGTTTTCCATTGTGTGCTAATGGACAAGTTTTTGGTTTATATAAAAATACATGTGTTGGTAGCGATAATGTTACTGACTTTAATGCAATTGCAACATGTGACTGGACAAATGCTGGTGATTACATTTTAGCTAACACCTGTACTGAAAGACTCAAGCTTTTTGCAGCAGAAACGCTCAAAGCTACTGAGGAGACATTTAAACTGTCTTATGGTATTGCTACTGTACGTGAAGTGCTGTCTGACAGAGAATTACATCTTTCATGGGAAGTTGGTAAACCTAGACCACCACTTAACCGAAATTATGTCTTTACTGGTTATCGTGTAACTAAAAACAGTAAAGTACAAATAGGAGAGTACACCTTTGAAAAAGGTGACTATGGTGATGCTGTTGTTTACCGAGGTACAACAACTTACAAATTAAATGTTGGTGATTATTTTGTGCTGACATCACATACAGTAATGCCATTAAGTGCACCTACACTAGTGCCACAAGAGCACTATGTTAGAATTACTGGCTTATACCCAACACTCAATATCTCAGATGAGTTTTCTAGCAATGTTGCAAATTATCAAAAGGTTGGTATGCAAAAGTATTCTACACTCCAGGGACCACCTGGTACTGGTAAGAGTCATTTTGCTATTGGCCTAGCTCTCTACTACCCTTCTGCTCGCATAGTGTATACAGCTTGCTCTCATGCCGCTGTTGATGCACTATGTGAGAAGGCATTAAAATATTTGCCTATAGATAAATGTAGTAGAATTATACCTGCACGTGCTCGTGTAGAGTGTTTTGATAAATTCAAAGTGAATTCAACATTAGAACAGTATGTCTTTTGTACTGTAAATGCATTGCCTGAGACGACAGCAGATATAGTTGTCTTTGATGAAATTTCAATGGCCACAAATTATGATTTGAGTGTTGTCAATGCCAGATTACGTGCTAAGCACTATGTGTACATTGGCGACCCTGCTCAATTACCTGCACCACGCACATTGCTAACTAAGGGCACACTAGAACCAGAATATTTCAATTCAGTGTGTAGACTTATGAAAACTATAGGTCCAGACATGTTCCTCGGAACTTGTCGGCGTTGTCCTGCTGAAATTGTTGACACTGTGAGTGCTTTGGTTTATGATAATAAGCTTAAAGCACATAAAGACAAATCAGCTCAATGCTTTAAAATGTTTTATAAGGGTGTTATCACGCATGATGTTTCATCTGCAATTAACAGGCCACAAATAGGCGTGGTAAGAGAATTCCTTACACGTAACCCTGCTTGGAGAAAAGCTGTCTTTATTTCACCTTATAATTCACAGAATGCTGTAGCCTCAAAGATTTTGGGACTACCAACTCAAACTGTTGATTCATCACAGGGCTCAGAATATGACTATGTCATATTCACTCAAACCACTGAAACAGCTCACTCTTGTAATGTAAACAGATTTAATGTTGCTATTACCAGAGCAAAAGTAGGCATACTTTGCATAATGTCTGATAGAGACCTTTATGACAAGTTGCAATTTACAAGTCTTGAAATTCCACGTAGGAATGTGGCAACTTTACAAGCTGAAAATGTAACAGGACTCTTTAAAGATTGTAGTAAGGTAATCACTGGGTTACATCCTACACAGGCACCTACACACCTCAGTGTTGACACTAAATTCAAAACTGAAGGTTTATGTGTTGACATACCTGGCATACCTAAGGACATGACCTATAGAAGACTCATCTCTATGATGGGTTTTAAAATGAATTATCAAGTTAATGGTTACCCTAACATGTTTATCACCCGCGAAGAAGCTATAAGACATGTACGTGCATGGATTGGCTTCGATGTCGAGGGGTGTCATGCTACTAGAGAAGCTGTTGGTACCAATTTACCTTTACAGCTAGGTTTTTCTACAGGTGTTAACCTAGTTGCTGTACCTACAGGTTATGTTGATACACCTAATAATACAGATTTTTCCAGAGTTAGTGCTAAACCACCGCCTGGAGATCAATTTAAACACCTCATACCACTTATGTACAAAGGACTTCCTTGGAATGTAGTGCGTATAAAGATTGTACAAATGTTAAGTGACACACTTAAAAATCTCTCTGACAGAGTCGTATTTGTCTTATGGGCACATGGCTTTGAGTTGACATCTATGAAGTATTTTGTGAAAATAGGACCTGAGCGCACCTGTTGTCTATGTGATAGACGTGCCACATGCTTTTCCACTGCTTCAGACACTTATGCCTGTTGGCATCATTCTATTGGATTTGATTACGTCTATAATCCGTTTATGATTGATGTTCAACAATGGGGTTTTACAGGTAACCTACAAAGCAACCATGATCTGTATTGTCAAGTCCATGGTAATGCACATGTAGCTAGTTGTGATGCAATCATGACTAGGTGTCTAGCTGTCCACGAGTGCTTTGTTAAGCGTGTTGACTGGACTATTGAATATCCTATAATTGGTGATGAACTGAAGATTAATGCGGCTTGTAGAAAGGTTCAACACATGGTTGTTAAAGCTGCATTATTAGCAGACAAATTCCCAGTTCTTCACGACATTGGTAACCCTAAAGCTATTAAGTGTGTACCTCAAGCTGATGTAGAATGGAAGTTCTATGATGCACAGCCTTGTAGTGACAAAGCTTATAAAATAGAAGAATTATTCTATTCTTATGCCACACATTCTGACAAATTCACAGATGGTGTATGCCTATTTTGGAATTGCAATGTCGATAGATATCCTGCTAATTCCATTGTTTGTAGATTTGACACTAGAGTGCTATCTAACCTTAACTTGCCTGGTTGTGATGGTGGCAGTTTGTATGTAAATAAACATGCATTCCACACACCAGCTTTTGATAAAAGTGCTTTTGTTAATTTAAAACAATTACCATTTTTCTATTACTCTGACAGTCCATGTGAGTCTCATGGAAAACAAGTAGTGTCAGATATAGATTATGTACCACTAAAGTCTGCTACGTGTATAACACGTTGCAATTTAGGTGGTGCTGTCTGTAGACATCATGCTAATGAGTACAGATTGTATCTCGATGCTTATAACATGATGATCTCAGCTGGCTTTAGCTTGTGGGTTTACAAACAATTTGATACTTATAACCTCTGGAACACTTTTACAAGACTTCAGAGTTTAGAAAATGTGGCTTTTAATGTTGTAAATAAGGGACACTTTGATGGACAACAGGGTGAAGTACCAGTTTCTATCATTAATAACACTGTTTACACAAAAGTTGATGGTGTTGATGTAGAATTGTTTGAAAATAAAACAACATTACCTGTTAATGTAGCATTTGAGCTTTGGGCTAAGCGCAACATTAAACCAGTACCAGAGGTGAAAATACTCAATAATTTGGGTGTGGACATTGCTGCTAATACTGTGATCTGGGACTACAAAAGAGATGCTCCAGCACATATATCTACTATTGGTGTTTGTTCTATGACTGACATAGCCAAGAAACCAACTGAAACGATTTGTGCACCACTCACTGTCTTTTTTGATGGTAGAGTTGATGGTCAAGTAGACTTATTTAGAAATGCCCGTAATGGTGTTCTTATTACAGAAGGTAGTGTTAAAGGTTTACAACCATCTGTAGGTCCCAAACAAGCTAGTCTTAATGGAGTCACATTAATTGGAGAAGCCGTAAAAACACAGTTCAATTATTATAAGAAAGTTGATGGTGTTGTCCAACAATTACCTGAAACTTACTTTACTCAGAGTAGAAATTTACAAGAATTTAAACCCAGGAGTCAAATGGAAATTGATTTCTTAGAATTAGCTATGGATGAATTCATTGAACGGTATAAATTAGAAGGCTATGCCTTCGAACATATCGTTTATGGAGATTTTAGTCATAGTCAGTTAGGTGGTTTACATCTACTGATTGGACTAGCTAAACGTTTTAAGGAATCACCTTTTGAATTAGAAGATTTTATTCCTATGGACAGTACAGTTAAAAACTATTTCATAACAGATGCGCAAACAGGTTCATCTAAGTGTGTGTGTTCTGTTATTGATTTATTACTTGATGATTTTGTTGAAATAATAAAATCCCAAGATTTATCTGTAGTTTCTAAGGTTGTCAAAGTGACTATTGACTATACAGAAATTTCATTTATGCTTTGGTGTAAAGATGGCCATGTAGAAACATTTTACCCAAAATTACAATCTAGTCAAGCGTGGCAACCGGGTGTTGCTATGCCTAATCTTTACAAAATGCAAAGAATGCTATTAGAAAAGTGTGACCTTCAAAATTATGGTGATAGTGCAACATTACCTAAAGGCATAATGATGAATGTCGCAAAATATACTCAACTGTGTCAATATTTAAACACATTAACATTAGCTGTACCCTATAATATGAGAGTTATACATTTTGGTGCTGGTTCTGATAAAGGAGTTGCACCAGGTACAGCTGTTTTAAGACAGTGGTTGCCTACGGGTACGCTGCTTGTCGATTCAGATCTTAATGACTTTGTCTCTGATGCAGATTCAACTTTGATTGGTGATTGTGCAACTGTACATACAGCTAATAAATGGGATCTCATTATTAGTGATATGTACGACCCTAAGACTAAAAATGTTACAAAAGAAAATGACTCTAAAGAGGGTTTTTTCACTTACATTTGTGGGTTTATACAACAAAAGCTAGCTCTTGGAGGTTCCGTGGCTATAAAGATAACAGAACATTCTTGGAATGCTGATCTTTATAAGCTCATGGGACACTTCGCATGGTGGACAGCCTTTGTTACTAATGTGAATGCGTCATCATCTGAAGCATTTTTAATTGGATGTAATTATCTTGGCAAACCACGCGAACAAATAGATGGTTATGTCATGCATGCAAATTACATATTTTGGAGGAATACAAATCCAATTCAGTTGTCTTCCTATTCTTTATTTGACATGAGTAAATTTCCCCTTAAATTAAGGGGTACTGCTGTTATGTCTTTAAAAGAAGGTCAAATCAATGATATGATTTTATCTCTTCTTAGTAAAGGTAGACTTATAATTAGAGAAAACAACAGAGTTGTTATTTCTAGTGATGTTCTTGTTAACAACTAAACGAACAATGTTTGTTTTTCTTGTTTTATTGCCACTAGTCTCTAGTCAGTGTGTTAATCTTACAACCAGAACTCAATTACCCCCTGCATACACTAATTCTTTCACACGTGGTGTTTATTACCCTGACAAAGTTTTCAGATCCTCAGTTTTACATTCAACTCAGGACTTGTTCTTACCTTTCTTTTCCAATGTTACTTGGTTCCATGCTATACATGTCTCTGGGACCAATGGTACTAAGAGGTTTGATAACCCTGTCCTACCATTTAATGATGGTGTTTATTTTGCTTCCACTGAGAAGTCTAACATAATAAGAGGCTGGATTTTTGGTACTACTTTAGATTCGAAGACCCAGTCCCTACTTATTGTTAATAACGCTACTAATGTTGTTATTAAAGTCTGTGAATTTCAATTTTGTAATGATCCATTTTTGGGTGTTTATTACCACAAAAACAACAAAAGTTGGATGGAAAGTGAGTTCAGAGTTTATTCTAGTGCGAATAATTGCACTTTTGAATATGTCTCTCAGCCTTTTCTTATGGACCTTGAAGGAAAACAGGGTAATTTCAAAAATCTTAGGGAATTTGTGTTTAAGAATATTGATGGTTATTTTAAAATATATTCTAAGCACACGCCTATTAATTTAGTGCGTGATCTCCCTCAGGGTTTTTCGGCTTTAGAACCATTGGTAGATTTGCCAATAGGTATTAACATCACTAGGTTTCAAACTTTACTTGCTTTACATAGAAGTTATTTGACTCCTGGTGATTCTTCTTCAGGTTGGACAGCTGGTGCTGCAGCTTATTATGTGGGTTATCTTCAACCTAGGACTTTTCTATTAAAATATAATGAAAATGGAACCATTACAGATGCTGTAGACTGTGCACTTGACCCTCTCTCAGAAACAAAGTGTACGTTGAAATCCTTCACTGTAGAAAAAGGAATCTATCAAACTTCTAACTTTAGAGTCCAACCAACAGAATCTATTGTTAGATTTCCTAATATTACAAACTTGTGCCCTTTTGGTGAAGTTTTTAACGCCACCAGATTTGCATCTGTTTATGCTTGGAACAGGAAGAGAATCAGCAACTGTGTTGCTGATTATTCTGTCCTATATAATTCCGCATCATTTTCCACTTTTAAGTGTTATGGAGTGTCTCCTACTAAATTAAATGATCTCTGCTTTACTAATGTCTATGCAGATTCATTTGTAATTAGAGGTGATGAAGTCAGACAAATCGCTCCAGGGCAAACTGGAAAGATTGCTGATTATAATTATAAATTACCAGATGATTTTACAGGCTGCGTTATAGCTTGGAATTCTAACAATCTTGATTCTAAGGTTGGTGGTAATTATAATTACCTGTATAGATTGTTTAGGAAGTCTAATCTCAAACCTTTTGAGAGAGATATTTCAACTGAAATCTATCAGGCCGGTAGCACACCTTGTAATGGTGTTGAAGGTTTTAATTGTTACTTTCCTTTACAATCATATGGTTTCCAACCCACTAATGGTGTTGGTTACCAACCATACAGAGTAGTAGTACTTTCTTTTGAACTTCTACATGCACCAGCAACTGTTTGTGGACCTAAAAAGTCTACTAATTTGGTTAAAAACAAATGTGTCAATTTCAACTTCAATGGTTTAACAGGCACAGGTGTTCTTACTGAGTCTAACAAAAAGTTTCTGCCTTTCCAACAATTTGGCAGAGACATTGCTGACACTACTGATGCTGTCCGTGATCCACAGACACTTGAGATTCTTGACATTACACCATGTTCTTTTGGTGGTGTCAGTGTTATAACACCAGGAACAAATACTTCTAACCAGGTTGCTGTTCTTTATCAGGATGTTAACTGCACAGAAGTCCCTGTTGCTATTCATGCAGATCAACTTACTCCTACTTGGCGTGTTTATTCTACAGGTTCTAATGTTTTTCAAACACGTGCAGGCTGTTTAATAGGGGCTGAACATGTCAACAACTCATATGAGTGTGACATACCCATTGGTGCAGGTATATGCGCTAGTTATCAGACTCAGACTAATTCTCCTCGGCGGGCACGTAGTGTAGCTAGTCAATCCATCATTGCCTACACTATGTCACTTGGTGCAGAAAATTCAGTTGCTTACTCTAATAACTCTATTGCCATACCCACAAATTTTACTATTAGTGTTACCACAGAAATTCTACCAGTGTCTATGACCAAGACATCAGTAGATTGTACAATGTACATTTGTGGTGATTCAACTGAATGCAGCAATCTTTTGTTGCAATATGGCAGTTTTTGTACACAATTAAACCGTGCTTTAACTGGAATAGCTGTTGAACAAGACAAAAACACCCAAGAAGTTTTTGCACAAGTCAAACAAATTTACAAAACACCACCAATTAAAGATTTTGGTGGTTTTAATTTTTCACAAATATTACCAGATCCATCAAAACCAAGCAAGAGGTCATTTATTGAAGATCTACTTTTCAACAAAGTGACACTTGCAGATGCTGGCTTCATCAAACAATATGGTGATTGCCTTGGTGATATTGCTGCTAGAGACCTCATTTGTGCACAAAAGTTTAACGGCCTTACTGTTTTGCCACCTTTGCTCACAGATGAAATGATTGCTCAATACACTTCTGCACTGTTAGCGGGTACAATCACTTCTGGTTGGACCTTTGGTGCAGGTGCTGCATTACAAATACCATTTGCTATGCAAATGGCTTATAGGTTTAATGGTATTGGAGTTACACAGAATGTTCTCTATGAGAACCAAAAATTGATTGCCAACCAATTTAATAGTGCTATTGGCAAAATTCAAGACTCACTTTCTTCCACAGCAAGTGCACTTGGAAAACTTCAAGATGTGGTCAACCAAAATGCACAAGCTTTAAACACGCTTGTTAAACAACTTAGCTCCAATTTTGGTGCAATTTCAAGTGTTTTAAATGATATCCTTTCACGTCTTGACAAAGTTGAGGCTGAAGTGCAAATTGATAGGTTGATCACAGGCAGACTTCAAAGTTTGCAGACATATGTGACTCAACAATTAATTAGAGCTGCAGAAATCAGAGCTTCTGCTAATCTTGCTGCTACTAAAATGTCAGAGTGTGTACTTGGACAATCAAAAAGAGTTGATTTTTGTGGAAAGGGCTATCATCTTATGTCCTTCCCTCAGTCAGCACCTCATGGTGTAGTCTTCTTGCATGTGACTTATGTCCCTGCACAAGAAAAGAACTTCACAACTGCTCCTGCCATTTGTCATGATGGAAAAGCACACTTTCCTCGTGAAGGTGTCTTTGTTTCAAATGGCACACACTGGTTTGTAACACAAAGGAATTTTTATGAACCACAAATCATTACTACAGACAACACATTTGTGTCTGGTAACTGTGATGTTGTAATAGGAATTGTCAACAACACAGTTTATGATCCTTTGCAACCTGAATTAGACTCATTCAAGGAGGAGTTAGATAAATATTTTAAGAATCATACATCACCAGATGTTGATTTAGGTGACATCTCTGGCATTAATGCTTCAGTTGTAAACATTCAAAAAGAAATTGACCGCCTCAATGAGGTTGCCAAGAATTTAAATGAATCTCTCATCGATCTCCAAGAACTTGGAAAGTATGAGCAGTATATAAAATGGCCATGGTACATTTGGCTAGGTTTTATAGCTGGCTTGATTGCCATAGTAATGGTGACAATTATGCTTTGCTGTATGACCAGTTGCTGTAGTTGTCTCAAGGGCTGTTGTTCTTGTGGATCCTGCTGCAAATTTGATGAAGACGACTCTGAGCCAGTGCTCAAAGGAGTCAAATTACATTACACATAAACGAACTTATGGATTTGTTTATGAGAATCTTCACAATTGGAACTGTAACTTTGAAGCAAGGTGAAATCAAGGATGCTACTCCTTCAGATTTTGTTCGCGCTACTGCAACGATACCGATACAAGCCTCACTCCCTTTCGGATGGCTTATTGTTGGCGTTGCACTTCTTGCTGTTTTTCAGAGCGCTTCCAAAATCATAACCCTCAAAAAGAGATGGCAACTAGCACTCTCCAAGGGTGTTCACTTTGTTTGCAACTTGCTGTTGTTGTTTGTAACAGTTTACTCACACCTTTTGCTCGTTGCTGCTGGCCTTGAAGCCCCTTTTCTCTATCTTTATGCTTTAGTCTACTTCTTGCAGAGTATAAACTTTGTAAGAATAATAATGAGGCTTTGGCTTTGCTGGAAATGCCGTTCCAAAAACCCATTACTTTATGATGCCAACTATTTTCTTTGCTGGCATACTAATTGTTACGACTATTGTATACCTTACAATAGTGTAACTTCTTCAATTGTCATTACTTCAGGTGATGGCACAACAAGTCCTATTTCTGAACATGACTACCAGATTGGTGGTTATACTGAAAAATGGGAATCTGGAGTAAAAGACTGTGTTGTATTACACAGTTACTTCACTTCAGACTATTACCAGCTGTACTCAACTCAATTGAGTACAGACACTGGTGTTGAACATGTTACCTTCTTCATCTACAATAAAATTGTTGATGAGCCTGAAGAACATGTCCAAATTCACACAATCGACGGTTCATCCGGAGTTGTTAATCCAGTAATGGAACCAATTTATGATGAACCGACGACGACTACTAGCGTGCCTTTGTAAGCACAAGCTGATGAGTACGAACTTATGTACTCATTCGTTTCGGAAGAGACAGGTACGTTAATAGTTAATAGCGTACTTCTTTTTCTTGCTTTCGTGGTATTCTTGCTAGTTACACTAGCCATCCTTACTGCGCTTCGATTGTGTGCGTACTGCTGCAATATTGTTAACGTGAGTCTTGTAAAACCTTCTTTTTACGTTTACTCTCGTGTTAAAAATCTGAATTCTTCTAGAGTTCCTGATCTTCTGGTCTAAACGAACTAAATATTATATTAGTTTTTCTGTTTGGAACTTTAATTTTAGCCATGGCAGATTCCAACGGTACTATTACCGTTGAAGAGCTTAAAAAGCTCCTTGAACAATGGAACCTAGTAATAGGTTTCCTATTCCTTACATGGATTTGTCTTCTACAATTTGCCTATGCCAACAGGAATAGGTTTTTGTATATAATTAAGTTAATTTTCCTCTGGCTGTTATGGCCAGTAACTTTAGCTTGTTTTGTGCTTGCTGCTGTTTACAGAATAAATTGGATCACCGGTGGAATTGCTATCGCAATGGCTTGTCTTGTAGGCTTGATGTGGCTCAGCTACTTCATTGCTTCTTTCAGACTGTTTGCGCGTACGCGTTCCATGTGGTCATTCAATCCAGAAACTAACATTCTTCTCAACGTGCCACTCCATGGCACTATTCTGACCAGACCGCTTCTAGAAAGTGAACTCGTAATCGGAGCTGTGATCCTTCGTGGACATCTTCGTATTGCTGGACACCATCTAGGACGCTGTGACATCAAGGACCTGCCTAAAGAAATCACTGTTGCTACATCACGAACGCTTTCTTATTACAAATTGGGAGCTTCGCAGCGTGTAGCAGGTGACTCAGGTTTTGCTGCATACAGTCGCTACAGGATTGGCAACTATAAATTAAACACAGACCATTCCAGTAGCAGTGACAATATTGCTTTGCTTGTACAGTAAGTGACAACAGATGTTTCATCTCGTTGACTTTCAGGTTACTATAGCAGAGATATTACTAATTATTATGAGGACTTTTAAAGTTTCCATTTGGAATCTTGATTACATCATAAACCTCATAATTAAAAATTTATCTAAGTCACTAACTGAGAATAAATATTCTCAATTAGATGAAGAGCAACCAATGGAGATTGATTAAACGAACATGAAAATTATTCTTTTCTTGGCACTGATAACACTCGCTACTTGTGAGCTTTATCACTACCAAGAGTGTGTTAGAGGTACAACAGTACTTTTAAAAGAACCTTGCTCTTCTGGAACATACGAGGGCAATTCACCATTTCATCCTCTAGCTGATAACAAATTTGCACTGACTTGCTTTAGCACTCAATTTGCTTTTGCTTGTCCTGACGGCGTAAAACACGTCTATCAGTTACGTGCCAGATCAGTTTCACCTAAACTGTTCATCAGACAAGAGGAAGTTCAAGAACTTTACTCTCCAATTTTTCTTATTGTTGCGGCAATAGTGTTTATAACACTTTGCTTCACACTCAAAAGAAAGACAGAATGATTGAACTTTCATTAATTGACTTCTATTTGTGCTTTTTAGCCTTTCTGCTATTCCTTGTTTTAATTATGCTTATTATCTTTTGGTTCTCACTTGAACTGCAAGATCATAATGAAACTTGTCACGCCTAAACGAACATGAAATTTCTTGTTTTCTTAGGAATCATCACAACTGTAGCTGCATTTCACCAAGAATGTAGTTTACAGTCATGTACTCAACATCAACCATATGTAGTTGATGACCCGTGTCCTATTCACTTCTATTCTAAATGGTATATTAGAGTAGGAGCTAGAAAATCAGCACCTTTAATTGAATTGTGCGTGGATGAGGCTGGTTCTAAATCACCCATTCAGTACATCGATATCGGTAATTATACAGTTTCCTGTTTACCTTTTACAATTAATTGCCAGGAACCTAAATTGGGTAGTCTTGTAGTGCGTTGTTCGTTCTATGAAGACTTTTTAGAGTATCATGACGTTCGTGTTGTTTTAGATTTCATCTAAACGAACAAACTAAAATGTCTGATAATGGACCCCAAAATCAGCGAAATGCACCCCGCATTACGTTTGGTGGACCCTCAGATTCAACTGGCAGTAACCAGAATGGAGAACGCAGTGGGGCGCGATCAAAACAACGTCGGCCCCAAGGTTTACCCAATAATACTGCGTCTTGGTTCACCGCTCTCACTCAACATGGCAAGGAAGACCTTAAATTCCCTCGAGGACAAGGCGTTCCAATTAACACCAATAGCAGTCCAGATGACCAAATTGGCTACTACCGAAGAGCTACCAGACGAATTCGTGGTGGTGACGGTAAAATGAAAGATCTCAGTCCAAGATGGTATTTCTACTACCTAGGAACTGGGCCAGAAGCTGGACTTCCCTATGGTGCTAACAAAGACGGCATCATATGGGTTGCAACTGAGGGAGCCTTGAATACACCAAAAGATCACATTGGCACCCGCAATCCTGCTAACAATGCTGCAATCGTGCTACAACTTCCTCAAGGAACAACATTGCCAAAAGGCTTCTACGCAGAAGGGAGCAGAGGCGGCAGTCAAGCCTCTTCTCGTTCCTCATCACGTAGTCGCAACAGTTCAAGAAATTCAACTCCAGGCAGCAGTAGGGGAACTTCTCCTGCTAGAATGGCTGGCAATGGCGGTGATGCTGCTCTTGCTTTGCTGCTGCTTGACAGATTGAACCAGCTTGAGAGCAAAATGTCTGGTAAAGGCCAACAACAACAAGGCCAAACTGTCACTAAGAAATCTGCTGCTGAGGCTTCTAAGAAGCCTCGGCAAAAACGTACTGCCACTAAAGCATACAATGTAACACAAGCTTTCGGCAGACGTGGTCCAGAACAAACCCAAGGAAATTTTGGGGACCAGGAACTAATCAGACAAGGAACTGATTACAAACATTGGCCGCAAATTGCACAATTTGCCCCCAGCGCTTCAGCGTTCTTCGGAATGTCGCGCATTGGCATGGAAGTCACACCTTCGGGAACGTGGTTGACCTACACAGGTGCCATCAAATTGGATGACAAAGATCCAAATTTCAAAGATCAAGTCATTTTGCTGAATAAGCATATTGACGCATACAAAACATTCCCACCAACAGAGCCTAAAAAGGACAAAAAGAAGAAGGCTGATGAAACTCAAGCCTTACCGCAGAGACAGAAGAAACAGCAAACTGTGACTCTTCTTCCTGCTGCAGATTTGGATGATTTCTCCAAACAATTGCAACAATCCATGAGCAGTGCTGACTCAACTCAGGCCTAAACTCATGCAGACCACACAAGGCAGATGGGCTATATAAACGTTTTCGCTTTTCCGTTTACGATATATAGTCTACTCTTGTGCAGAATGAATTCTCGTAACTACATAGCACAAGTAGATGTAGTTAACTTTAATCTCACATAGCAATCTTTAATCAGTGTGTAACATTAGGGAGGACTTGAAAGAGCCACCACATTTTCACCGAGGCCACGCGGAGTACGATCGAGTGTACAGTGAACAATGCTAGGGAGAGCTGCCTATATGGAAGAGCCCTAATGTGTAAAATTAATTTTAGTAGTGCTATCCCCATGTGATTTTAATAGCTTCTTAGGAGAATGACAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAAA
Now we have reached the end of the example, here are some questions for you to try. You might need to add a few more cells below so that you can use the files and Python that we used earlier to help answer them: