Purpose. Barcode amplification for Ion Torrent. Barcode amplification for lon Torrent. Barcode amplification for lon Torrent. Barcode amplification for lon Torrent.
Supplementary Data for Pettitt S.J. et al “Genome-wide barcoded transposon screen for cancer drug sensitivity in haploid mouse embryonic stem cells” ! ! Contents:! ! 1! Supplementary!Table!1!–!62TG!resistant!mutants! 2! PCR!conditions! 3–4! Oligonucleotide!sequences! 5–10! Barcoded!transposon!plasmid!sequence!in!GenBank!format! !
Supplementary9Table91:+Full+Sanger+sequencing+mapping+data+for+6DTG+resistant+colonies Note+that+transfection+conditions+were+not+optimised+for+singleDcopy+insertions+in+this+ screen,+therefore+there+are+multiple+insertions+for+many+clones. Colonies 13 5 4 4 2 2 2 2 2 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1
PB5+chr 17 17 17 NA 8 18 17 2 13 17 17 17 17 17 17 17 17 5 17 NA NA NA NA NA 12 2 8 2 4 2 10 5 NA NA NA NA NA NA NA NA NA NA NA 15 5 19 15 3 10 2 15 6 13
PB5+pos 87,980,842 14,075,902 87,981,979 NA 15,983,713 46,607,977 87,695,676 66,181,060 80,932,417 87,696,147 87,686,487 87,977,132 87,976,095 87,986,894 87,985,785 87,986,027 87,977,320 143,913,337 87,975,970 NA NA NA NA NA 105,216,775 4,575,487 24,626,040 91,018,524 125,610,100 155,202,987 42,394,530 45,495,981 NA NA NA NA NA NA NA NA NA NA NA 81,397,057 116,076,073 26,828,382 84,191,907 38,420,211 30,193,691 118,176,835 93,632,521 49,236,495 94,902,797
PB3+chr 17 17 17 17 2 7 NA 2 3 17 NA 11 17 17 17 19 2 NA 12 9 17 17 17 17 17 17 11 15 3 NA 9 8 X 17 4 1 14 5 X 14 6 17 10 NA 16 8 NA NA NA 4 2 19 13
PB3+pos PB5+gene 87,980,841 Msh6% 87,976,095 87,981,978 Msh6% 87,673,130 NA 30,885,944 Csmd1& 72,326,037 Eif1a& NA Msh2% 66,181,064 7,641,626 87,696,154 Msh2% NA Msh2% 11,871,662 Msh6% 30,602,248 Msh6% 87,986,950 Msh6% 87,985,784 Msh6% 47,789,915 Msh6% 166,481,574 Msh6% NA Pms2% 27,420,386 Msh6% 111,270,757 NA 87,976,106 NA 87,982,198 NA 87,982,154 NA 87,986,026 NA 87,708,692 Tcl1& 87,976,094 Frmd4a& 107,448,585 Adam18& 53,929,745 Celf1& 41,607,029 Grik3& NA Itch& 57,048,835 Lace1& 71,239,563 Lap3& 144,362,807 NA 16,702,867 NA 4,133,834 NA 189,730,157 NA 63,494,809 NA 116,076,026 NA 8,433,292 NA 14,314,940 NA 112,537,455 NA 78,948,231 NA 82,175,718 NA NA St13& 33,405,964 Tmem233& 94,081,687 NA NA NA 121,199,306 85,902,464 43,920,226 94,903,088
PB3+gene Msh6% Msh2%
NA
NA
NA Mlh1% Msh6% Msh6% Msh6% Msh6%
NA
Alg13& NA Penk& Ptpn14& Tdh& Tmem233&
NA Bbs2& NA NA NA Rlf&
Supplementary,Data,.,PCR,conditions Barcode,PCR Temperature)(°C) Time)(s) Cycles 98 30 98 60 72
10 10 25)cycles 10
72
60
Inverse,PCR,(Primary,and,secondary) Temperature)(°C) Time)(s) Cycles 98 30 98 65 72
10 10 18)cycles 30
72
60
Indexing,PCR Temperature)(°C) Time)(s) Cycles 98 30 51.7 72 72
30 30 6)cycles 5
Oligonucleotide sequences Name Sims_ion_1 Sims_ion_2 Sims_ion_3 Sims_ion_4 Sims_ion_5 Sims_ion_6 Sims_ion_7 Sims_ion_8 Sims_ion_9 Sims_ion_10 Sims_ion_11 Sims_ion_12 Sims_ion_13 Sims_ion_14 Sims_ion_15 Sims_ion_16 West_ion_1 West_ion_2 West_ion_3 West_ion_4 West_ion_5 West_ion_6 West_ion_7 West_ion_8 West_ion_9 West_ion_10 West_ion_11 West_ion_12
Sequence CCATCTCATCCCTGCGTGTCTCCGACTCAGCTAAGGTAACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTAAGGAGAACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGAAGAGGATTCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTACCAAGATCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGCAGAAGGAACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGCTGCAAGTTCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTTCGTGATTCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTTCCGATAACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTGAGCGGAACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGCTGACCGAACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTCCTCGAATCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTAGGTGGTTCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTAACGGACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTTGGAGTGTCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTAGAGGTCgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTGGATGACgatCTAAAGTAGCCCCTTGAATTC CCATCTCATCCCTGCGTGTCTCCGACTCAGCTAAGGTAACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTAAGGAGAACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGAAGAGGATTCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTACCAAGATCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGCAGAAGGAACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGCTGCAAGTTCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTTCGTGATTCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTTCCGATAACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTGAGCGGAACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGCTGACCGAACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTCCTCGAATCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTAGGTGGTTCgatCTAAAGCGCATGCTCCAGACTG
Purpose Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent
West_ion_13 West_ion_14 West_ion_15 West_ion_16 Sims_ion_R West_ion_R PB3_inv_R1 PB3_inv_F1 PB5_inv_F1 PB5_inv_R1 PB5_illu_F PB3_illu_F PB5_illu_R_full PB3_illu_R_full Ewsr1_gRNA_F Ewsr1_gRNA_R Ewsr1-F Ewsr1-R
CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTAACGGACgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTTGGAGTGTCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTAGAGGTCgatCTAAAGCGCATGCTCCAGACTG CCATCTCATCCCTGCGTGTCTCCGACTCAGTCTGGATGACgatCTAAAGCGCATGCTCCAGACTG CCTCTCTATGGGCAGTCGGTGATCCTCAGCTAGTGAAGCCACAGATGTA CCTCTCTATGGGCAGTCGGTGATCCTCAGCCTTGAACCTCCTCGTTCGACC CTCTTCTGCACTTGAACCTCCTC TTCAAGAATGCATGCGTCAATTTT GATATACAGACCGATAAAACACATGCGTCA AGTCAGAAACAACTTTGGCCTAAA ACACTCTTTCCCTACACGACGCTCTTCCGATCTGTACGTCACAATATGATTATCTTTCTAG ACACTCTTTCCCTACACGACGCTCTTCCGATCTGTCAATTTTACGCAGACTATCTTTCTAG gtgactggagttcAGACGTGTGCTCTTCCGATCtGCCTAAAGTAGCCCCTTGAATTC gtgactggagttcAGACGTGTGCTCTTCCGATCtCTTCTGCACTTGAACCTCCTCGTT CACCgTATGCAGTCTGCCCGTAGG AAACCCTACGGGCAGACTGCATAc AAGCAGCTCAGTTTAAGAAGGC CAAGGTGCTGGCTGAATCTTG
Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Barcode amplification for Ion Torrent Inverse PCR (first round) Inverse PCR (first round) Inverse PCR (first round) Inverse PCR (first round) Inverse PCR for HiSeq (second round) Inverse PCR for HiSeq (second round) Inverse PCR for HiSeq (second round) Inverse PCR for HiSeq (second round) Ewsr1 CRISPR vector cloning Ewsr1 CRISPR vector cloning Ewsr1 CRISPR genotyping Ewsr1 CRISPR genotyping
LOCUS pPB_SA_IRES_Neo_ 6152 bp ds-DNA circular 06-AUG-2013 DEFINITION . ACCESSION VERSION SOURCE . ORGANISM . COMMENT COMMENT ApEinfo:methylated:0 FEATURES Location/Qualifiers primer_bind complement(3317..3336) /label=T3 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_feature 2868..3177 /label=PB3 /ApEinfo_fwdcolor=#0cfeff /ApEinfo_revcolor=#0cfeff /ApEinfo_graphicformat=arrow_data {{} {0 1 2 0 0 -1} 0} width 5 offset 0 primer_bind 6106..6123 /label=M13-fwd /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_feature 530..966 /note="the human bcl-2 gene splice acceptor (intron 2 modified exon 3)" /label=SA /ApEinfo_fwdcolor=#ff80ed /ApEinfo_revcolor=#ffcc66 /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer_bind complement(3354..3374) /label=M13-rev /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 misc_feature 58..320 /label=PB5 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 primer_bind 6132..6152 /label=T7 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 promoter 1573..1610 /label=gb3 /ApEinfo_fwdcolor=#1014ff /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data {{0 1 2 0 0 -1} {} 0} width 5 offset 0 rep_origin complement(3742..4424)
misc_feature
rep_origin
misc_feature
CDS
misc_feature
misc_binding
promoter
CDS
misc_feature
/label=ColE1 origin /ApEinfo_fwdcolor=gray50 /ApEinfo_revcolor=gray50 /ApEinfo_graphicformat=arrow_data width 5 offset 0 2799..2821 /label=primer-right /ApEinfo_fwdcolor=#ff0006 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 complement(5642..5948) /label=F1 ori /ApEinfo_fwdcolor=gray50 /ApEinfo_revcolor=gray50 /ApEinfo_graphicformat=arrow_data width 5 offset 0 785..787 /note="an artificial stop codon" /label=an artificial stop codon /ApEinfo_fwdcolor=#ff8000 /ApEinfo_revcolor=#ffcc66 /ApEinfo_graphicformat=arrow_data width 5 offset 0 complement(5967..6035) /label=LacZ alpha /ApEinfo_fwdcolor=#6495ed /ApEinfo_revcolor=#6495ed /ApEinfo_graphicformat=arrow_data width 5 offset 0 416..437 /label=primer-Left /ApEinfo_fwdcolor=#ff0612 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 complement(3380..3402) /label=LacO /ApEinfo_fwdcolor=#6495ed /ApEinfo_revcolor=#6495ed /ApEinfo_graphicformat=arrow_data width 5 offset 0 980..1559 /label=ires /ApEinfo_fwdcolor=#ff970a /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 complement(4522..5181) /label=AmpR /ApEinfo_fwdcolor=yellow /ApEinfo_revcolor=yellow /ApEinfo_graphicformat=arrow_data width 5 offset 0 438..458 /label=p5+mir3 /ApEinfo_fwdcolor=#32ff82 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{} {0 1 2 0 0 -1} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
CDS
misc_feature
polyA_site
misc_feature
misc_feature
primer_bind
misc_feature
misc_feature
CDS
misc_feature
1611..2457 /label=neo /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 484..502 /label=p7+Loop /ApEinfo_fwdcolor=#07ffa6 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 2580..2701 /label=SV40 polyA /ApEinfo_fwdcolor=#0fff34 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 2731..2752 /label=PB3seqF /ApEinfo_fwdcolor=#00ff4d /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 2778..2798 /label=PB3seq_R /ApEinfo_fwdcolor=#11ff66 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 2898..2925 /label=PB3_seq_R /ApEinfo_fwdcolor=#ffe609 /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 459..483 /label=PB5 barcode /ApEinfo_fwdcolor=#f111ff /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data 0 2753..2777 /label=PB3 barcode /ApEinfo_fwdcolor=#dc1dff /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data 0 1666..2457 /label=Kan/neoR /ApEinfo_fwdcolor=yellow /ApEinfo_revcolor=yellow /ApEinfo_graphicformat=arrow_data width 5 offset 0 116..145 /label=PB5_inv_F1 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0
{{0 1 2 0 0 -1} {} 0}
{{} {0 1 2 0 0 -1} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{} {0 1 2 0 0 -1} 0}
{{} {0 1 2 0 0 -1} 0}
{{} {} 0} width 5 offset
{{} {} 0} width 5 offset
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
misc_feature
misc_feature
misc_feature
misc_feature
misc_feature
misc_feature
misc_feature
83..110 /label=PB5_inv_F2 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 419..442 /label=PB5_inv_R1 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 436..458 /label=PB5_inv_R2 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 2786..2808 /label=PB3_inv_F1 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 3129..3152 /label=PB3_inv_F2 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 2783..2806 /label=PB3_inv_R1 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0 3153..3176 /label=PB3_inv_R2 /ApEinfo_fwdcolor=cyan /ApEinfo_revcolor=green /ApEinfo_graphicformat=arrow_data width 5 offset 0
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
{{0 1 2 0 0 -1} {} 0}
ORIGIN 1 61 121 181 241 301 361 421 481 541 601 661 721 781 841 901 961
CGAATTGGCG ACCCTAGAAA CATGTGTTTT TTATTATATT TTATTTATTA ACATTCTCTC ataaaataag TCAGAAACAA NNNTACATCT TCTAAGatgg gtggtttctg ggtgcccagg tctctaatac actgtaaggc gctcagtttg aagtcaacat ATGCATCCGC
GTATTCACGA GATAATCATA ATCGGTCTGT TACACTTACA AAAAAAAACA TTTTACAAAA taattagctt CTTTGGCCTA GTggcttcac ctggcacgta aatcatggcc ctgccctggc tctttctttt cccagcatgc gccctggtgg gcctgcccca GGTGGTAAGA
CAGCAGGCTG TTGTGACGTA ATATCGAGGT TACTAATAAT AAAACTCAAA ataaacttat aacctataca AAGTAGCCCC tagctagcgc aagagggtct tccaaagagc aggatgcctc tcttccctcc ggcctctgtt gagcttgcat aacaaatatg ATTCCGCCCC
AATAATAAAA CGTTAAAGAT TTATTTATTA AAATTCAACA ATTTCTTCTA tttgtacttt taatagaaac TTGAATTCNN attagttatt caataaatat atttgccttt tttctctgtg tctcccccaa tgatttctcc caccctgggt caaaaggttc TCTCCCTCCC
AAATTAGAAA AATCATGCGT ATTTGAATAG AACAATTTAT TAAAGTAACA aaaaacagtc aaattatact NNNNNNNNNN aatccgggat gccttgaatt tcccctctca gggtggcatt ctgcaggatg tggctgtctc gcctatctgg aCTAAAGCAG CCCCCCCTAA
CTATTATTTA AAAATTGACG ATATTAAGTT TTATGTTTAT AAACTTTTAA atgttgtatt tattaGTCAG NNNNNNNNNN cTGCTGTATC aatgaaaaca cccttctgaa ctctgctctc cctttgtgga tgaagactct gccacaagtg TAGAAATAAT CGTTACTGGC
1021 1081 1141 1201 1261 1321 1381 1441 1501 1561 1621 1681 1741 1801 1861 1921 1981 2041 2101 2161 2221 2281 2341 2401 2461 2521 2581 2641 2701 2761 2821 2881 2941 3001 3061 3121 3181 3241 3301 3361 3421 3481 3541 3601 3661 3721 3781 3841 3901 3961 4021 4081 4141 4201 4261 4321 4381 4441 4501 4561
CGAAGCCGCT CCGTCTTTTG AGGGGTCTTT GTTCCTCTGG AACCCCCCAC GCAAAGGCGG TGGCTCTCCT ATGGGATCTG AACGTCTAGG ATGGCCACAA TCTGCTTACA TTGCACGCAG CAGACAATCG CTTTTTGTCA CTATCGTGGC GCGGGAAGGG CTTGCTCCTG GATCCGGCTA CGGATGGAAG CCAGCCGAAC ACCCATGGCG ATCGACTGTG GATATTGCTG GCCGCTCCCG GGCCGCGACT AAAACCTCCC ACTTGTTTAT ATAAAGCATT AAGGCGTAAA NNNNNNNNNN Gctcatcgtc AAATATATAT ATGAGTTAAA CGTTATAGTT AAGCGGCGAC AGCAATATTT AGATTTGCGC ACCACATTGT GCGAGCCAGC ATAGCTGTTT AAGCATAAAG GCGCTCACTG CCAACGCGCG CTCGCTGCGC ACGGTTATCC AAAGGCCAGG TGACGAGCAT AAGATACCAG GCTTACCGGA ACGCTGTAGG ACCCCCCGTT GGTAAGACAC GTATGTAGGC GACAGTATTT CTCTTGATCC GATTACGCGC CGCTCAGTGG CTTCACCTAG GTAAACTTGG TCTATTTCGT
TGGAATAAGG GCAATGTGAG CCCCTCTCGC AAGCTTCTTG CTGGCGACAG CACAACCCCA CAAGCGTATT ATCTGGGGCC CCCCCCGAAC CCTGTTACAT TAAACAGTAA GTTCTCCGGC GCTGCTCTGA AGACCGACCT TGGCCACGAC ACTGGCTGCT CCGAGAAAGT CCTGCCCATT CCGGTCTTGT TGTTCGCCAG ATGCCTGCTT GCCGGCTGGG AAGAGCTTGG ATTCGCAGCG CTAGATCATA ACACCTCCCC TGCAGCTTAT TTTTTCACTG TTGTAAGGGT NNNNNNNGGT taaagaacta ATAATAAgtt TCTTAAAAGT CAAAATCAGT TGAGATGTCC CAAGAATGCA TTTACTCGAC GGCCAAGCTG TTTTGTTCCC CCTGTGTGAA TGTAAAGCCT CCCGCTTTCC GGGAGAGGCG TCGGTCGTTC ACAGAATCAG AACCGTAAAA CACAAAAATC GCGTTTCCCC TACCTGTCCG TATCTCAGTT CAGCCCGACC GACTTATCGC GGTGCTACAG GGTATCTGCG GGCAAACAAA AGAAAAAAAG AACGAAAACT ATCCTTTTAA TCTGACAGTT TCATCCATAG
CCGGTGTGCG GGCCCGGAAA CAAAGGAATG AAGACAAACA GTGCCTCTGC GTGCCACGTT CAACAAGGGG TCGGTGCACA CACGGGGACG TGCACAAGAT TACAAGGGGT CGCTTGGGTG TGCCGCCGTG GTCCGGTGCC GGGCGTTCCT ATTGGGCGAA ATCCATCATG CGACCACCAA CGATCAGGAT GCTCAAGGCG GCCGAATATC TGTGGCGGAC CGGCGAATGG CATCGCCTTC ATCAGCCATA CTGAACCTGA AATGGTTACA CATTCTAGTT ACCAGGATCC CGAACGAGGA cccattttat atcacgtaag CACGTAAAAG GACACTTACC TAAATGCACA TGCGTCAATT CTAAACTTTA TGTGACGCGA TTTAGTGAGG ATTGTTATCC GGGGTGCCTA AGTCGGGAAA GTTTGCGTAT GGCTGCGGCG GGGATAACGC AGGCCGCGTT GACGCTCAAG CTGGAAGCTC CCTTTCTCCC CGGTGTAGGT GCTGCGCCTT CACTGGCAGC AGTTCTTGAA CTCTGCTGAA CCACCGCTGG GATCTCAAGA CACGTTAAGG ATTAAAAATG ACCAATGCTT TTGCCTGACT
TTTGTCTATA CCTGGCCCTG CAAGGTCTGT ACGTCTGTAG GGCCAAAAGC GTGAGTTGGA CTGAAGGATG TGCTTTACAT TGGTTTTCCT AAAAATATAT GTTATgggat GAGAGGCTAT TTCCGGCTGT CTGAATGAAC TGCGCAGCTG GTGCCGGGGC GCTGATGCAA GCGAAACATC GATCTGGACG CGCATGCCCG ATGGTGGAAA CGCTATCAGG GCTGACCGCT TATCGCCTTC CCACATTTGT AACATAAAAT AATAAAGCAA GTGGTTtgtc CTAAAGCGCA GGTTCAAGTG tatatattag tagaacatga ATAATCATGC GCATTGACAA GCGACGGATT TTACGCAGAC AACACGTCAT CGCGCGCTAA GTTAATTGCG GCTCACAATT ATGAGTGAGC CCTGTCGTGC TGGGCGCTCT AGCGGTATCA AGGAAAGAAC GCTGGCGTTT TCAGAGGTGG CCTCGTGCGC TTCGGGAAGC CGTTCGCTCC ATCCGGTAAC AGCCACTGGT GTGGTGGCCT GCCAGTTACC TAGCGGTGGT AGATCCTTTG GATTTTGGTC AAGTTTTAAA AATCAGTGAG CCCCGTCGTG
TGTTATTTTC TCTTCTTGAC TGAATGTCGT CGACCCTTTG CACGTGTATA TAGTTGTGGA CCCAGAAGGT GTGTTTAGTC TTGAAAAACA CATCACGAAC cggccattga TCGGCTATGA CAGCGCAGGG TGCAGGACGA TGCTCGACGT AGGATCTCCT TGCGGCGGCT GCATCGAGCG AAGAGCATCA ACGGCGAGGA ATGGCCGCTT ACATAGCGTT TCCTCGTGCT TTGACGAGTT AGAGGTTTTA GAATGCAATT TAGCATCACA caaactcatc TGCTCCAGAC CAGAAGAGCA tcacgatATC aataaCAATA GTCATTTTGA GCACGCCTCA CGCGCTATTT TATCTTTCTA AGAATCTTCG AGAATGGCAA CGCTTGGCGT CCACACAACA TAACTCACAT CAGCTGCATT TCCGCTTCCT GCTCACTCAA ATGTGAGCAA TTCCATAGGC CGAAACCCGA TCTCCTGTTC GTGGCGCTTT AAGCTGGGCT TATCGTCTTG AACAGGATTA AACTACGGCT TTCGGAAAAA TTTTTTGTTT ATCTTTTCTA ATGAGATTAT TCAATCTAAA GCACCTATCT TAGATAACTA
CACCATATTG GAGCATTCCT GAAGGAAGCA CAGGCAGCGG AGATACACCT AAGAGTCAAA ACCCCATTGT GAGGTTAAAA CGATGATAAT AGTAAAACTG acaagATGGA CTGGGCACAA GCGCCCGGTT GGCAGCGCGG TGTCACTGAA GTCATCTCAC GCATACGCTT AGCACGTACT GGGGCTCGCG TCTCGTCGTG TTCTGGATTC GGCTACCCGT TTACGGTATC CTTCTGAAGC CTTGCTTTAA GTTGTTGTTA AATTTCACAA aatgtatcTT TGNNNNNNNN GAGAGGATAT TATAACAAGA TAATTATCGT CTCACGCGGT CGGGAGCTCC AGAAAGAGAG GGGTTAAAAA TTTGACAAAA ACCAAGTCGC AATCATGGTC TACGAGCCGG TAATTGCGTT AATGAATCGG CGCTCACTGA AGGCGGTAAT AAGGCCAGCA TCCGCCCCCC CAGGACTATA CGACCCTGCC CTCATAGCTC GTGTGCACGA AGTCCAACCC GCAGAGCGAG ACACTAGAAG GAGTTGGTAG GCAAGCAGCA CGGGGTCTGA CAAAAAGGAT GTATATATGA CAGCGATCTG CGATACGGGA
4621 4681 4741 4801 4861 4921 4981 5041 5101 5161 5221 5281 5341 5401 5461 5521 5581 5641 5701 5761 5821 5881 5941 6001 6061 6121 //
GGGCTTACCA AGATTTATCA TTTATCCGCC AGTTAATAGT GTTTGGTATG CATGTTGTGC GGCCGCAGTG ATCCGTAAGA TATGCGGCGA CAGAACTTTA CTTACCGCTG ATCTTTTACT AAAGGGAATA TTGAAGCATT AAATAAACAA GTTAATATTT TAGGCCGAAA GTTGTTCCAG CGAAAAACCG TTGGGGTCGA GCTTGACGGG GGCGCTAGGG CTTAATGCGC GGGCGATCGG AGGCGATTAA AGTGAGCGCG
TCTGGCCCCA GCAATAAACC TCCATCCAGT TTGCGCAACG GCTTCATTCA AAAAAAGCGG TTATCACTCA TGCTTTTCTG CCGAGTTGCT AAAGTGCTCA TTGAGATCCA TTCACCAGCG AGGGCGACAC TATCAGGGTT ATAGGGGTTC TGTTAAAATT TCGGCAAAAT TTTGGAACAA TCTATCAGGG GGTGCCGTAA GAAAGCCGGC CGCTGGCAAG CGCTACAGGG TGCGGGCCTC GTTGGGTAAC CGTAATACGA
GTGCTGCAAT AGCCAGCCGG CTATTAATTG TTGTTGCCAT GCTCCGGTTC TTAGCTCCTT TGGTTATGGC TGACTGGTGA CTTGCCCGGC TCATTGGAAA GTTCGATGTA TTTCTGGGTG GGAAATGTTG ATTGTCTCAT CGCGCACATT CGCGTTAAAT CCCTTATAAA GAGTCCACTA CGATGGCCCA AGCACTAAAT GAACGTGGCG TGTAGCGGTC CGCGTCCCAT TTCGCTATTA GCCAGGGTTT CTCACTATAG
GATACCGCGA AAGGGCCGAG TTGCCGGGAA TGCTACAGGC CCAACGATCA CGGTCCTCCG AGCACTGCAT GTACTCAACC GTCAATACGG ACGTTCTTCG ACCCACTCGT AGCAAAAACA AATACTCATA GAGCGGATAC TCCCCGAAAA TTTTGTTAAA TCAAAAGAAT TTAAAGAACG CTACGTGAAC CGGAACCCTA AGAAAGGAAG ACGCTGCGCG TCGCCATTCA CGCCAGCTGG TCCCAGTCAC GG
GACCCACGCT CGCAGAAGTG GCTAGAGTAA ATCGTGGTGT AGGCGAGTTA ATCGTTGTCA AATTCTCTTA AAGTCATTCT GATAATACCG GGGCGAAAAC GCACCCAACT GGAAGGCAAA CTCTTCCTTT ATATTTGAAT GTGCCACCTA TCAGCTCATT AGACCGAGAT TGGACTCCAA CATCACCCTA AAGGGAGCCC GGAAGAAAGC TAACCACCAC GGCTGCGCAA CGAAAGGGGG GACGTTGTAA
CACCGGCTCC GTCCTGCAAC GTAGTTCGCC CACGCTCGTC CATGATCCCC GAAGTAAGTT CTGTCATGCC GAGAATAGTG CGCCACATAG TCTCAAGGAT GATCTTCAGC ATGCCGCAAA TTCAATATTA GTATTTAGAA AATTGTAAGC TTTTAACCAA AGGGTTGAGT CGTCAAAGGG ATCAAGTTTT CCGATTTAGA GAAAGGAGCG ACCCGCCGCG CTGTTGGGAA ATGTGCTGCA AACGACGGCC
