| 质粒名称: | p201N Cas9 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 59175 |
| 存储实验室: | Wayne Parrott |
|
载体骨架基本信息 |
|
| 载体名称: | pPZP |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 6200 bp |
| 完整质粒大小: | 14349 bp |
| 载体修饰: | 利用aph基因对卡那霉素进行细菌筛选。 |
| 载体类型: | CRISPR |
| 筛选标记: | G418,卡那霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9,Cas9 human-codon optimized |
| 物种来源: | Synthetic |
| 插入大小: | 5265 bp |
| 启动子 | 2x35S |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | I-SceI(未销毁) |
| 3' 克隆位点: | I-SceI(未销毁) |
| 5' 测序引物 | ACATGCACCTAATTTCACTAGATGT |
| 3' 测序引物 | TGTAAAACGACGGCCAGT |
|
文献引用方法 |
|
| 材料和方法部分: | p201N Cas9 was a gift from Wayne Parrott (Addgene plasmid # 59175 ; http://n2t.net/addgene:59175 ; RRID:Addgene_59175) |
| 参考文献部分: | Targeted genome modifications in soybean with CRISPR/Cas9. Jacobs TB, LaFayette PR, Schmitz RJ, Parrott WA. . BMC Biotechnology. 2015;15:16 10.1186/s12896-015-0131-2 |
p201N Cas9
价格:元
规格: 2ug质粒(仅用于转化提取) 转染级去内毒素液体即用质粒100μg 转染级去内毒素液体即用质粒500μg 转染级去内毒素液体即用质粒1mg 更大规格-质粒构建-质粒定制-请联系我们
联系方式:I47-825O-882O
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p201N Cas9载体 p201NCas9质粒基本信息
p201N Cas9质粒图谱 p201NCas9载体图谱和p201N Cas9载体序列 p201NCas9质粒序列多克隆位点信息

p201N Cas9质粒 p201NCas9载体简介
p201N Cas9质粒序列 p201NCas9载体序列
LOCUS p201N Cas9 14414 bp ds-DNA circular SYN 18-DEC-2019
DEFINITION Cas9 driven by double 35S, nptII for plant selection, I-PpoI site to
accept gRNA from pUC gRNA Shuttle.
ACCESSION .
VERSION .
KEYWORDS p201N Cas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14414)
AUTHORS Jacobs TB, LaFayette PR, Schmitz RJ, Parrott WA.
TITLE Targeted genome modifications in soybean with CRISPR/Cas9.
JOURNAL BMC Biotechnology. 2015;15:16
REFERENCE 2 (bases 1 to 14414)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Dec 18, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14414
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 1217..1846
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 2275..3348
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 3414..3608
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(3844..3866)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 3952..4092
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 4007..4026
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(4107..4124)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(4278..4866)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(4358..4377)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(4961..5776)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 5684..5703
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 5768..5787
/label=pENTR-R
/note="pENTR vectors, reverse primer"
misc_feature 6069..6093
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
protein_bind 6270..6291
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 6306..6336
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6344..6360
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
primer_bind 6349..6371
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 6368..6384
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 6368..6384
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS complement(6906..7700)
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7017..7036)
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
primer_bind 7627..7646
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
promoter 8846..9522
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 9459..9479
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 9568..13671
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 13684..13704
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
terminator 13771..14023
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(14064..14081)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(14064..14080)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(14073..14095)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 14276..14300
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
ORIGIN
1 agtactttaa agtactttaa agtactttaa agtactttga tccaacccct ccgctgctat
61 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag
121 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt
181 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag
241 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc
301 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc
361 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac
421 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt
481 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac
541 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag
601 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg
661 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc
721 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc
781 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt
841 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc
901 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg
961 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct
1021 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc
1081 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg
1141 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa
1201 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa
1261 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt
1321 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc
1381 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg
1441 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat
1501 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac
1561 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc
1621 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc
1681 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc
1741 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg
1801 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa
1861 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc
1921 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag
1981 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt
2041 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat
2101 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac
2161 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg
2221 ggttgtctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc
2281 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag
2341 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt
2401 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc
2461 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg
2521 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt
2581 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac
2641 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg
2701 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag
2761 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat
2821 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt
2881 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa
2941 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag
3001 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg
3061 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg
3121 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc
3181 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca
3241 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc
3301 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag
3361 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat
3421 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac
3481 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa
3541 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc
3601 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg
3661 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca
3721 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca
3781 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga
3841 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
3901 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat
3961 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag
4021 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa
4081 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
4141 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
4201 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
4261 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
4321 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
4381 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
4441 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
4501 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
4561 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
4621 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
4681 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct
4741 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
4801 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
4861 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
4921 cgttaaggga ttttggtcat gcaggatcat gaattaattc ttagaaaaac tcatcgagca
4981 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
5041 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
5101 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
5161 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
5221 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
5281 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
5341 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
5401 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
5461 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
5521 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
5581 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
5641 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
5701 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
5761 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
5821 ttcatgatct ggatcacagg cagcaacgct ctgtcatcgt tacaatcaac atgctaccct
5881 ccgcgagatc atccgtgttt caaacccggc agcttagttg ccgttcttcc gaatagcatc
5941 ggtaacatga gcaaagtctg ccgccttaca acggctctcc cgctgacgcc gtcccggact
6001 gatgggctgc ctgtatcgag tggtgatttt gtgccgagct gccggtcggg gagctgttgg
6061 ctggctggtg gcaggatata ttgtggtgta aacaaattga cgcttagaca acttaataac
6121 acattgcgga cgtttttaat gtactgaatt aacgccgaat tgctctagcc aatacgcaaa
6181 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
6241 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
6301 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
6361 tttcacacag gaaacagcta tgacatgatt acgaatcctt gactagaggg taggcgcggc
6421 ttaattaagc gcttaattaa gcgcttaatt aagcgcttaa taagccgcgc cgtaccccaa
6481 tagtctcgac agacacatag cacctaacat tattgcttaa atttactatt tttttactat
6541 attatactca acccaatgag cataaagact gtaaaatctc aaattcctga gaagcatatt
6601 tatcgatccc acagacttga tagttccata atccatacgc tgcagccaaa ttgctagtgt
6661 gttgaacatt taacacgtag agaactagaa aagatataaa actaagattg atatccaaaa
6721 tagacgagaa caataagcaa aaactcttag ttttgaaata aatcaacaat cccgagggtt
6781 gtcacataca tcaaaaacga aaatccatat agcaaaaaaa actctaaatt accgttcgac
6841 aaaaagagaa aactgatagg acatttgcta aacattaaaa tcaaaatttg gatcaagccg
6901 aattctcaga agaactcgtc aagaaggcga tagaaggcga tgcgctgcga atcgggagcg
6961 gcgataccgt aaagcacgag gaagcggtca gcccattcgc cgccaagctc ttcagcaata
7021 tcacgggtag ccaacgctat gtcctgatag cggtccgcca cacccagccg gccacagtcg
7081 atgaatccag aaaagcggcc attttccacc atgatattcg gcaagcaggc atcgccatgg
7141 gtcacgacga gatcatcgcc gtcgggcatg cgcgccttga gcctggcgaa cagttcggct
7201 ggcgcgagcc cctgatgctc ttcgtccaga tcatcctgat cgacaagacc ggcttccatc
7261 cgagtacgtg ctcgctcgat gcgatgtttc gcttggtggt cgaatgggca ggtagccgga
7321 tcaagcgtat gcagccgccg cattgcatca gccatgatgg atactttctc ggcaggagca
7381 aggtgagatg acaggagatc ctgccccggc acttcgccca atagcagcca gtcccttccc
7441 gcttcagtga caacgtcgag cacagctgcg caaggaacgc ccgtcgtggc cagccacgat
7501 agccgcgctg cctcgtcctg cagttcattc agggcaccgg acaggtcggt cttgacaaaa
7561 agaaccgggc gcccctgcgc tgacagccgg aacacggcgg catcagagca gccgattgtc
7621 tgttgtgccc agtcatagcc gaatagcctc tccacccaag cggccggaga acctgcgtgc
7681 aatccatctt gttcaatcat ctcgagacgt accgcgcgat gcattcgaag atcgatcctt
7741 cgcctggagg agagaaatca gtggcgctgc ggcttttagg gtttctttgt tgatggaatg
7801 agagtgtaag ctctgccagt gccactttat tagggtttta caagcccttt tcttcgtaat
7861 tgggcctgac attttgtgcc acttgggcct ttagagatga aaatgtatat tgggcttaag
7921 ttgacttgaa ggataaatta gtttaggata ttacgttttt tatgagaatt ggtgtgtcgg
7981 atacatgtat atgatgcatt caaatatatg tattctagat acatttaagt ttagatacaa
8041 tctaaaatgt gtctttaatt acaggactgt aactaaaata cttaatgtaa gaagaatatt
8101 actcctttaa tagcttttga gtatatctag tctaacatct tttaaaaaag tctaatttct
8161 ttcatttatt tttcgagcaa tagcaaagtg cataattatt tttttcttct agaaattcag
8221 atttgtttct ctaaattttg agattctttt ctcaattttg tatgtctaga gaacaatgtg
8281 tatttttcac tctagttggt tgttactttg ttgaatgttc tgataaaagt atattgttat
8341 ttctgaagta gatataaacc ttcatttgga aattatacat aaatcaaaat cgttaattat
8401 ctagatcaag atatatgccc ttttcctaat gtatttgata catgcaccta atttcactag
8461 atgtatcttt tctatttttt aaattatgaa tagttaattt tttccatatg tgtatttgat
8521 acatacttca tgactttaaa aaattaatta tataccagat atatgtattt aaaatttgtt
8581 atgtatttaa agtatgtata tgattattcg atattaatct cttcgatgaa atttaaatcg
8641 ataactatgt gctttggatc gatctgccca ctagtgagtc gtattacaat tcatgactct
8701 cttaaggtag ccaaaatcga tatctaggga taacagggta atcgatcact agaattcggc
8761 gcgccgggcc caacatggtg gagcacgaca ctctcgtcta ctccaagaat atcaaagata
8821 cagtctcaga agaccaaagg gctattgaga cttttcaaca aagggtaata tcgggaaacc
8881 tcctcggatt ccattgccca gctatctgtc acttcatcaa aaggacagta gaaaaggaag
8941 gtggcaccta caaatgccat cattgcgata aaggaaaggc tatcgttcaa gatgcctctg
9001 ccgacagtgg tcccaaagat ggacccccac ccacgaggag catcgtggaa aaagaagacg
9061 ttccaaccac gtcttcaaag caagtggatt gatgtgaaca tggtggagca cgacactctc
9121 gtctactcca agaatatcaa agatacagtc tcagaagacc aaagggctat tgagactttt
9181 caacaaaggg taatatcggg aaacctcctc ggattccatt gcccagctat ctgtcacttc
9241 atcaaaagga cagtagaaaa ggaaggtggc acctacaaat gccatcattg cgataaagga
9301 aaggctatcg ttcaagatgc ctctgccgac agtggtccca aagatggacc cccacccacg
9361 aggagcatcg tggaaaaaga agacgttcca accacgtctt caaagcaagt ggattgatgt
9421 gatatctcca ctgacgtaag ggatgacgca caatcccact atccttcgca agacccttcc
9481 tctatataag gaagttcatt tcatttggag aggacacgct gaaatcacca gtctctctct
9541 acaaatctat ctctctcgac cgctagcatg gacaagaagt actccattgg gctcgatatc
9601 ggcacaaaca gcgtcggctg ggccgtcatt acggacgagt acaaggtgcc gagcaaaaaa
9661 ttcaaagttc tgggcaatac cgatcgccac agcataaaga agaacctcat tggcgccctc
9721 ctgttcgact ccggggagac ggccgaagcc acgcggctca aaagaacagc acggcgcaga
9781 tatacccgca gaaagaatcg gatctgctac ctgcaggaga tctttagtaa tgagatggct
9841 aaggtggatg actctttctt ccataggctg gaggagtcct ttttggtgga ggaggataaa
9901 aagcacgagc gccacccaat ctttggcaat atcgtggacg aggtggcgta ccatgaaaag
9961 tacccaacca tatatcatct gaggaagaag cttgtagaca gtactgataa ggctgacttg
10021 cggttgatct atctcgcgct ggcgcatatg atcaaatttc ggggacactt cctcatcgag
10081 ggggacctga acccagacaa cagcgatgtc gacaaactct ttatccaact ggttcagact
10141 tacaatcagc ttttcgaaga gaacccgatc aacgcatccg gagttgacgc caaagcaatc
10201 ctgagcgcta ggctgtccaa atcccggcgg ctcgaaaacc tcatcgcaca gctccctggg
10261 gagaagaaga acggcctgtt tggtaatctt atcgccctgt cactcgggct gacccccaac
10321 tttaaatcta acttcgacct ggccgaagat gccaagcttc aactgagcaa agacacctac
10381 gatgatgatc tcgacaatct gctggcccag atcggcgacc agtacgcaga cctttttttg
10441 gcggcaaaga acctgtcaga cgccattctg ctgagtgata ttctgcgagt gaacacggag
10501 atcaccaaag ctccgctgag cgctagtatg atcaagcgct atgatgagca ccaccaagac
10561 ttgactttgc tgaaggccct tgtcagacag caactgcctg agaagtacaa ggaaattttc
10621 ttcgatcagt ctaaaaatgg ctacgccgga tacattgacg gcggagcaag ccaggaggaa
10681 ttttacaaat ttattaagcc catcttggaa aaaatggacg gcaccgagga gctgctggta
10741 aagcttaaca gagaagatct gttgcgcaaa cagcgcactt tcgacaatgg aagcatcccc
10801 caccagattc acctgggcga actgcacgct atcctcaggc ggcaagagga tttctacccc
10861 tttttgaaag ataacaggga aaagattgag aaaatcctca catttcggat accctactat
10921 gtaggccccc tcgcccgggg aaattccaga ttcgcgtgga tgactcgcaa atcagaagag
10981 accatcactc cctggaactt cgaggaagtc gtggataagg gggcctctgc ccagtccttc
11041 atcgaaagga tgactaactt tgataaaaat ctgcctaacg aaaaggtgct tcctaaacac
11101 tctctgctgt acgagtactt cacagtttat aacgagctca ccaaggtcaa atacgtcaca
11161 gaagggatga gaaagccagc attcctgtct ggagagcaga agaaagctat cgtggacctc
11221 ctcttcaaga cgaaccggaa agttaccgtg aaacagctca aagaagacta tttcaaaaag
11281 attgaatgtt tcgactctgt tgaaatcagc ggagtggagg atcgcttcaa cgcatccctg
11341 ggaacgtatc acgatctcct gaaaatcatt aaagacaagg acttcctgga caatgaggag
11401 aacgaggaca ttcttgagga cattgtcctc acccttacgt tgtttgaaga tagggagatg
11461 attgaagaac gcttgaaaac ttacgctcat ctcttcgacg acaaagtcat gaaacagctc
11521 aagaggcgcc gatatacagg atgggggcgg ctgtcaagaa aactgatcaa tgggatccga
11581 gacaagcaga gtggaaagac aatcctggat tttcttaagt ccgatggatt tgccaaccgg
11641 aacttcatgc agttgatcca tgatgactct ctcaccttta aggaggacat ccagaaagca
11701 caagtttctg gccaggggga cagtcttcac gagcacatcg ctaatcttgc aggtagccca
11761 gctatcaaaa agggaatact gcagaccgtt aaggtcgtgg atgaactcgt caaagtaatg
11821 ggaaggcata agcccgagaa tatcgttatc gagatggccc gagagaacca aactacccag
11881 aagggacaga agaacagtag ggaaaggatg aagaggattg aagagggtat aaaagaactg
11941 gggtcccaaa tccttaagga acacccagtt gaaaacaccc agcttcagaa tgagaagctc
12001 tacctgtact acctgcagaa cggcagggac atgtacgtgg atcaggaact ggacatcaat
12061 cggctctccg actacgacgt ggatcatatc gtgccccagt cttttctcaa agatgattct
12121 attgataata aagtgttgac aagatccgat aaaaatagag ggaagagtga taacgtcccc
12181 tcagaagaag ttgtcaagaa aatgaaaaat tattggcggc agctgctgaa cgccaaactg
12241 atcacacaac ggaagttcga taatctgact aaggctgaac gaggtggcct gtctgagttg
12301 gataaagccg gcttcatcaa aaggcagctt gttgagacac gccagatcac caagcacgtg
12361 gcccaaattc tcgattcacg catgaacacc aagtacgatg aaaatgacaa actgattcga
12421 gaggtgaaag ttattactct gaagtctaag ctggtctcag atttcagaaa ggactttcag
12481 ttttataagg tgagagagat caacaattac caccatgcgc atgatgccta cctgaatgca
12541 gtggtaggca ctgcacttat caaaaaatat cccaagcttg aatctgaatt tgtttacgga
12601 gactataaag tgtacgatgt taggaaaatg atcgcaaagt ctgagcagga aataggcaag
12661 gccaccgcta agtacttctt ttacagcaat attatgaatt ttttcaagac cgagattaca
12721 ctggccaatg gagagattcg gaagcgacca cttatcgaaa caaacggaga aacaggagaa
12781 atcgtgtggg acaagggtag ggatttcgcg acagtccgga aggtcctgtc catgccgcag
12841 gtgaacatcg ttaaaaagac cgaagtacag accggaggct tctccaagga aagtatcctc
12901 ccgaaaagga acagcgacaa gctgatcgca cgcaaaaaag attgggaccc caagaaatac
12961 ggcggattcg attctcctac agtcgcttac agtgtactgg ttgtggccaa agtggagaaa
13021 gggaagtcta aaaaactcaa aagcgtcaag gaactgctgg gcatcacaat catggagcga
13081 tcaagcttcg aaaaaaaccc catcgacttt ctcgaggcga aaggatataa agaggtcaaa
13141 aaagacctca tcattaagct tcccaagtac tctctctttg agcttgaaaa cggccggaaa
13201 cgaatgctcg ctagtgcggg cgagctgcag aaaggtaacg agctggcact gccctctaaa
13261 tacgttaatt tcttgtatct ggccagccac tatgaaaagc tcaaagggtc tcccgaagat
13321 aatgagcaga agcagctgtt cgtggaacaa cacaaacact accttgatga gatcatcgag
13381 caaataagcg aattctccaa aagagtgatc ctcgccgacg ctaacctcga taaggtgctt
13441 tctgcttaca ataagcacag ggataagccc atcagggagc aggcagaaaa cattatccac
13501 ttgtttactc tgaccaactt gggcgcgcct gcagccttca agtacttcga caccaccata
13561 gacagaaagc ggtacacctc tacaaaggag gtcctggacg ccacactgat tcatcagtca
13621 attacggggc tctatgaaac aagaatcgac ctctctcagc tcggtggaga cagcagggct
13681 gaccccaaga agaagaggaa ggtgtgaccg cggatatctg cagaagctta cgcgtcgacg
13741 tctcgaggcc ggccgagctc gaatttcccc gatcgttcaa acatttggca ataaagtttc
13801 ttaagattga atcctgttgc cggtcttgcg atgattatca tataatttct gttgaattac
13861 gttaagcatg taataattaa catgtaatgc atgacgttat ttatgagatg ggtttttatg
13921 attagagtcc cgcaattata catttaatac gcgatagaaa acaaaatata gcgcgcaaac
13981 taggataaat tatcgcgcgc ggtgtcatct atgttactag atcgggaatt gatatctagg
14041 gataacaggg taataagctt ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
14101 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
14161 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatga
14221 gcttgagctt ggatcagatt gtcgtttccc gccttcagtt taaactatca gtgtttgaca
14281 ggatatattg gcgggtaaac ctaagagaaa agagcgttta ttagaataat cggatattta
14341 aaagggcgtg aaaaggttta tccgttcgtc catttgtatg tgcatgccaa ccacagggtt
14401 cccctcggga tcaa
//

