| 质粒名称: | pWS158 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 90517 |
| 存储实验室: | Tom Ellis |
|
载体骨架基本信息 |
|
| 载体名称: | 2 micron |
| 载体出品公司: | Clontech |
| 空载体大小: | 3831bp |
| 完整质粒大小: | 11081bp |
| 载体类型: | Yeast Expression, CRISPR |
| 载体抗性: | Kanamycin, 50 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | S. cerevisiae (budding yeast) |
|
文献引用方法 |
|
| 材料和方法部分: | pWS158 was a gift from Tom Ellis (Addgene plasmid # 90517 ; http://n2t.net/addgene:90517 ; RRID:Addgene_90517) |
买家导航
pWS158载体质粒基本信息
pWS158质粒图谱载体图谱和pWS158载体序列质粒序列多克隆位点信息
pWS158质粒载体简介
pWS158质粒序列载体序列
LOCUS pWS158 11081 bp DNA circular SYN 14-APR-2025
DEFINITION Cas9 gap repair vector - URA3.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11081)
TITLE Ellis Lab Yeast CRISPR Plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 11081)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT Sequence Label: pWS158
FEATURES Location/Qualifiers
source 1..11081
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 1..589
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 490..509
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 1451..5554
/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 1451..5554
/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 5567..5587
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5591..5611
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
RBS 6521..6532
/note="strong bacterial ribosome binding site (Elowitz and
Leibler, 2000)"
CDS 6539..7252
/codon_start=1
/product="GFP variant that folds robustly even when fused
to poorly folded proteins (Pedelacq et al., 2006)"
/label=superfolder GFP
/translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
FVTAAGITHGMDELYK"
terminator 7264..7335
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 7351..7378
/label=T7Te terminator
/note="phage T7 early transcription terminator"
misc_RNA 7392..7467
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
CDS 8116..8919
/codon_start=1
/gene="S. cerevisiae URA3"
/product="orotidine-5'-phosphate decarboxylase, required
for uracil biosynthesis"
/label=URA3
/note="yeast auxotrophic marker, counterselectable with
5-fluoroorotic acid (5-FOA)"
/translation="MSKATYKERAATHPSPVAAKLFNIMHEKQTNLCASLDVRTTKELL
ELVEALGPKICLLKTHVDILTDFSMEGTVKPLKALSAKYNFLLFEDRKFADIGNTVKLQ
YSAGVYRIAEWADITNAHGVVGPGIVSGLKQAAEEVTKEPRGLLMLAELSCKGSLSTGE
YTKGTVDIAKSDKDFVIGFIAQRDMGGRDEGYDWLIMTPGVGLDDKGDALGQQYRTVDD
VVSTGSDIIIVGRGLFAKGRDAKVEGERYRKAGWEAYLRRCGQQN"
rep_origin 9002..9984
/label=2u ori
/note="yeast 2u plasmid origin of replication"
CDS complement(10042..10857)
/codon_start=1
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
CDS complement(10042..10857)
/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 10765..10784
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 10849..10868
/label=pENTR-R
/note="pENTR vectors, reverse primer"
ORIGIN
1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
661 gttatcccct gattctgtgg ataaccgtgc ggccgcccct gaattcgcat ctagactgaa
721 ctggccgata attgcagacg aacggtgagt aaggaaagag tgaggaacta tcgcatacct
781 gcatttaaag atgccgattt gggcgcgaat cctttatttt ggcttcaccc tcatactatt
841 atcagggcca gaaaaaggaa gtgtttccct ccttcttgaa ttgatgttac cctcataaag
901 cacgtggcct cttatcgaga aagaaattac cgtcgctcgt gatttgtttg caaaaagaac
961 aaaactgaaa aaacccagac acgctcgact tcctgtcatc ctattgattg cagcttccaa
1021 tttcgtcaca caacaaggtc ctagcgacgg ctcacaggtt ttgtaacaag caatcgaagg
1081 ttctggaatg gcgggaaagg gtttagtacc acatgctatg atgcccactg tgatctccag
1141 agcaaagttc gttcgatcgt actgttactc tctctctttc aaacagaatt gtccgaatcg
1201 tgtgacaaca acagcctgtt ctcacacact cttttcttct aaccaagggg gtggtttagt
1261 ttagtagaac ctcgtgaaac ttacatttac atatatataa acttgcataa attggtcaat
1321 gcaagaaata catatttggt cttttctaat tcgtagtttt tcaagttctt agatgctttc
1381 tttttctctt ttttacagat catcaaggaa gtaattatct actttttaca acaaatataa
1441 aacaagatct atggacaaga agtattctat cggactggat atcgggacta atagcgtcgg
1501 gtgggccgtg atcactgacg agtacaaggt gccctctaag aagttcaagg tgctcgggaa
1561 caccgaccgg cattccatca agaaaaatct gatcggagct ctcctctttg attcagggga
1621 aaccgctgaa gcaacccgcc tcaagcggac tgctagacgg cggtacacca ggaggaagaa
1681 ccggatttgt taccttcaag agatattctc caacgaaatg gcaaaggtcg acgacagctt
1741 cttccatagg ctggaagaat cattcctcgt ggaagaggat aagaagcatg aacggcatcc
1801 catcttcggt aatatcgtcg acgaggtggc ctatcacgag aaatacccaa ccatctacca
1861 tcttcgcaaa aagctggtgg actcaaccga caaggcagac ctccggctta tctacctggc
1921 cctggcccac atgatcaagt tcagaggcca cttcctgatc gagggcgacc tcaatcctga
1981 caatagcgat gtggataaac tgttcatcca gctggtgcag acttacaacc agctctttga
2041 agagaacccc atcaatgcaa gcggagtcga tgccaaggcc attctgtcag cccggctgtc
2101 aaagagccgc agacttgaga atcttatcgc tcagctgccg ggtgaaaaga aaaatggact
2161 gttcgggaac ctgattgctc tttcacttgg gctgactccc aatttcaagt ctaatttcga
2221 cctggcagag gatgccaagc tgcaactgtc caaggacacc tatgatgacg atctcgacaa
2281 cctcctggcc cagatcggtg accaatacgc cgaccttttc cttgctgcta agaatctttc
2341 tgacgccatc ctgctgtctg acattctccg cgtgaacact gaaatcacca aggcccctct
2401 ttcagcttca atgattaagc ggtatgatga gcaccaccag gacctgaccc tgcttaaggc
2461 actcgtccgg cagcagcttc cggagaagta caaggaaatc ttctttgacc agtcaaagaa
2521 tggatacgcc ggctacatcg acggaggtgc ctcccaagag gaattttata agtttatcaa
2581 acctatcctt gagaagatgg acggcaccga agagctcctc gtgaaactga atcgggagga
2641 tctgctgcgg aagcagcgca ctttcgacaa tgggagcatt ccccaccaga tccatcttgg
2701 ggagcttcac gccatccttc ggcgccaaga ggacttctac ccctttctta aggacaacag
2761 ggagaagatt gagaaaattc tcactttccg catcccctac tacgtgggac ccctcgccag
2821 aggaaatagc cggtttgctt ggatgaccag aaagtcagaa gaaactatca ctccctggaa
2881 cttcgaagag gtggtggaca agggagccag cgctcagtca ttcatcgaac ggatgactaa
2941 cttcgataag aacctcccca atgagaaggt cctgccgaaa cattccctgc tctacgagta
3001 ctttaccgtg tacaacgagc tgaccaaggt gaaatatgtc accgaaggga tgaggaagcc
3061 cgcattcctg tcaggcgaac aaaagaaggc aattgtggac cttctgttca agaccaatag
3121 aaaggtgacc gtgaagcagc tgaaggagga ctatttcaag aaaattgaat gcttcgactc
3181 tgtggagatt agcggggtcg aagatcggtt caacgcaagc ctgggtactt accatgatct
3241 gcttaagatc atcaaggaca aggattttct ggacaatgag gagaacgagg acatccttga
3301 ggacattgtc ctgactctca ctctgttcga ggaccgggaa atgatcgagg agaggcttaa
3361 gacctacgcc catctgttcg acgataaagt gatgaagcaa cttaaacgga gaagatatac
3421 cggatgggga cgccttagcc gcaaactcat caacggaatc cgggacaaac agagcggaaa
3481 gaccattctt gatttcctta agagcgacgg attcgctaat cgcaacttca tgcaacttat
3541 ccatgatgat tccctgacct ttaaggagga catccagaag gcccaagtgt ctggacaagg
3601 tgactcactg cacgagcata tcgcaaatct ggctggttca cccgctatta agaagggtat
3661 tctccagacc gtgaaagtcg tggacgagct ggtcaaggtg atgggtcgcc ataaaccaga
3721 gaacattgtc atcgagatgg ccagggaaaa ccagactacc cagaagggac agaagaacag
3781 cagggagcgg atgaaaagaa ttgaggaagg gattaaggag ctcgggtcac agatccttaa
3841 agagcacccg gtggaaaaca cccagcttca gaatgagaag ctctatctgt actaccttca
3901 aaatggacgc gatatgtatg tggaccaaga gcttgatatc aacaggctct cagactacga
3961 cgtggaccat atcgtccctc agagcttcct caaagacgac tcaattgaca ataaggtgct
4021 gactcgctca gacaagaacc ggggaaagtc agataacgtg ccctcagagg aagtcgtgaa
4081 aaagatgaag aactattggc gccagcttct gaacgcaaag ctgatcactc agcggaagtt
4141 cgacaatctc actaaggctg agaggggcgg actgagcgaa ctggacaaag caggattcat
4201 taaacggcaa cttgtggaga ctcggcagat tactaaacat gtcgcccaaa tccttgactc
4261 acgcatgaat accaagtacg acgaaaacga caaacttatc cgcgaggtga aggtgattac
4321 cctgaagtcc aagctggtca gcgatttcag aaaggacttt caattctaca aagtgcggga
4381 gatcaataac tatcatcatg ctcatgacgc atatctgaat gccgtggtgg gaaccgccct
4441 gatcaagaag tacccaaagc tggaaagcga gttcgtgtac ggagactaca aggtctacga
4501 cgtgcgcaag atgattgcca aatctgagca ggagatcgga aaggccaccg caaagtactt
4561 cttctacagc aacatcatga atttcttcaa gaccgaaatc acccttgcaa acggtgagat
4621 ccggaagagg ccgctcatcg agactaatgg ggagactggc gaaatcgtgt gggacaaggg
4681 cagagatttc gctaccgtgc gcaaagtgct ttctatgcct caagtgaaca tcgtgaagaa
4741 aaccgaggtg caaaccggag gcttttctaa ggaatcaatc ctccccaagc gcaactccga
4801 caagctcatt gcaaggaaga aggattggga ccctaagaag tacggcggat tcgattcacc
4861 aactgtggct tattctgtcc tggtcgtggc taaggtggaa aaaggaaagt ctaagaagct
4921 caagagcgtg aaggaactgc tgggtatcac cattatggag cgcagctcct tcgagaagaa
4981 cccaattgac tttctcgaag ccaaaggtta caaggaagtc aagaaggacc ttatcatcaa
5041 gctcccaaag tatagcctgt tcgaactgga gaatgggcgg aagcggatgc tcgcctccgc
5101 tggcgaactt cagaagggta atgagctggc tctcccctcc aagtacgtga atttcctcta
5161 ccttgcaagc cattacgaga agctgaaggg gagccccgag gacaacgagc aaaagcaact
5221 gtttgtggag cagcataagc attatctgga cgagatcatt gagcagattt ccgagttttc
5281 taaacgcgtc attctcgctg atgccaacct cgataaagtc cttagcgcat acaataagca
5341 cagagacaaa ccaattcggg agcaggctga gaatatcatc cacctgttca ccctcaccaa
5401 tcttggtgcc cctgccgcat tcaagtactt cgacaccacc atcgaccgga aacgctatac
5461 ctccaccaaa gaagtgctgg acgccaccct catccaccag agcatcaccg gactttacga
5521 aactcggatt gacctctcac agctcggagg ggatgaggga gctgatccaa aaaagaagag
5581 aaaggtagat ccaaaaaaga agagaaaggt ataaggatcc taactcgaga ttgaattgaa
5641 ttgaaatcga tagatcaatt tttttctttt ctctttcccc atcctttacg ctaaaataat
5701 agtttatttt attttttgaa tattttttat ttatatacgt atatatagac tattatttat
5761 cttttaatga ttattaagat ttttattaaa aaaaaattcg ctcctctttt aatgccttta
5821 tgcagttttt ttttcccatt cgatatttct atgtgctgcc aagatatcaa caacagagga
5881 catatgccct acctccatga tctactgacg tccctgaggc tgcaattcat gtaatggggc
5941 agtatccgcg gcaagtccta gtgcaatggc ggttttttac cctcgttctg aagaagaggc
6001 gacgcgggtg cggtcatcac taatgtggaa attggtaaga ctctcgggcc tccgccttta
6061 ggcggtgctt actctttcat aaaggggctg ttagttatgc aacgatgtgc ttcagtatta
6121 cattttttgc cttcaacgcc ttgattgttc tatttttgct aataataaat ctatttcatc
6181 ggactaaaag tccattagtt gtaagcggat ttagctcagt tgggagagcg ccagactgaa
6241 gaaaaacttc ggtcaagtca tctggaggtc ctgtgttcga tccacagaat tcgcagatgg
6301 ccggcatggt cccagcctcc tcgctggcgc cggctgggca acaccttcgg gtggcgaatg
6361 ggacttgaga cggaaagtga aacgtgattt catgcgtcat tttgaacatt ttgtaaatct
6421 tatttaataa tgtgtgcggc aattcacatt taatttatga atgttttctt aacatcgcgg
6481 caactcaaga aacggcaggt tcggatctta gctactagag aaagaggaga aatactagat
6541 gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac tggatggtga
6601 tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa ctaatggtaa
6661 actgacgctg aagttcatct gtactactgg taaactgccg gttccttggc cgactctggt
6721 aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata tgaagcagca
6781 tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga tttcctttaa
6841 ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata ccctggtaaa
6901 ccgcattgag ctgaaaggca ttgactttaa agaggacggc aatatcctgg gccataagct
6961 ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa aaaatggcat
7021 taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc tggctgatca
7081 ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca atcactatct
7141 gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata tggttctgct
7201 ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca aatgaccagg
7261 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
7321 tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc tttctgcgtt
7381 tatacgtctc agttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa
7441 cttgaaaaag tggcaccgag tcggtgcttt ttttattttt tgtcactatt gttatgtaaa
7501 atgccacctc tgacagtatg gaacgcaaac ttctgtctag tggatagctg gacattccag
7561 gcagtgcctc tgccgccgga cccctctcgt gattgggtag ctggacatgc ccttgtaaga
7621 tataacaaga gcctgcctgt ctaatgatct cacggcgaaa gtcggggaga cagcagcggc
7681 tgcagacatt ataccgcaac aacactaagg tgagataact ccgtacattg actacgcgtt
7741 cctctagacc ttacttgacc ggatacagtg tctttgacac gtttatgggt tacagcaatc
7801 acatccaaga ctggctatgc acgaagcaac tcttgagtgt taaaatgttg acccctgtat
7861 ttgggatgcg ggtagtagat gagtgcagga tatcagcagt tacagagatg ttacgaacca
7921 ctagtgcact gcagtacacg gtttccttga aatttttttg attcggtaat ctccgaacag
7981 aaggaagaac gaaggaagga gcacagactt agattggtat atatacgcat atgtagtgtt
8041 gaagaaacat gaaattgccc agtattctta acccaactgc acagaacaaa aacgtgcagg
8101 aaacgaagat aaatcatgtc gaaagctaca tataaggaac gtgctgctac tcatcctagt
8161 cctgttgctg ccaagctatt taatatcatg cacgaaaagc aaacaaactt gtgtgcttca
8221 ttggatgttc gtaccaccaa ggaattactg gagttagttg aagcattagg tcccaaaatt
8281 tgtttactaa aaacacatgt ggatatcttg actgattttt ccatggaggg cacagttaag
8341 ccgctaaagg cattatccgc caagtacaat tttttactct tcgaggacag aaaatttgct
8401 gacattggta atacagtcaa attgcagtac tctgcgggtg tatacagaat agcagaatgg
8461 gcagacatta cgaatgcaca cggtgtggtg ggcccaggta ttgttagcgg tttgaagcag
8521 gcggcagaag aagtaacaaa ggaacctaga ggccttttga tgttagcaga attgtcatgc
8581 aagggctccc tatctactgg agaatatact aagggtactg ttgacattgc gaagagcgac
8641 aaagattttg ttatcggctt tattgctcaa agagacatgg gtggaagaga tgaaggttac
8701 gattggttga ttatgacacc cggtgtgggt ttagatgaca agggagatgc attgggtcaa
8761 cagtatagaa ccgtggatga tgtggtttct acaggatctg acattattat tgttggaaga
8821 ggactatttg caaagggaag ggatgctaag gtagagggtg aacgttacag aaaagcaggc
8881 tgggaagcat atttgagaag atgcggccag caaaactaaa aaactgtatt ataagtaaat
8941 gcatgtatac taaactcaca aattagagct tcaatttaat tatatcagtt attacccgag
9001 taacgaagca tctgtgcttc attttgtaga acaaaaatgc aacgcgagag cgctaatttt
9061 tcaaacaaag aatctgagct gcatttttac agaacagaaa tgcaacgcga aagcgctatt
9121 ttaccaacga agaatctgtg cttcattttt gtaaaacaaa aatgcaacgc gagagcgcta
9181 atttttcaaa caaagaatct gagctgcatt tttacagaac agaaatgcaa cgcgagagcg
9241 ctattttacc aacaaagaat ctatacttct tttttgttct acaaaaatgc atcccgagag
9301 cgctattttt ctaacaaagc atcttagatt actttttttc tcctttgtgc gctctataat
9361 gcagtctctt gataactttt tgcactgtag gtccgttaag gttagaagaa ggctactttg
9421 gtgtctattt tctcttccat aaaaaaagcc tgactccact tcccgcgttt actgattact
9481 agcgaagctg cgggtgcatt ttttcaagat aaaggcatcc ccgattatat tctataccga
9541 tgtggattgc gcatactttg tgaacagaaa gtgatagcgt tgatgattct tcattggtca
9601 gaaaattatg aacggtttct tctattttgt ctctatatac tacgtatagg aaatgtttac
9661 attttcgtat tgttttcgat tcactctatg aatagttctt actacaattt ttttgtctaa
9721 agagtaatac tagagataaa cataaaaaat gtagaggtcg agtttagatg caagttcaag
9781 gagcgaaagg tggatgggta ggttatatag ggatatagca cagagatata tagcaaagag
9841 atacttttga gcaatgtttg tggaagcggt attcgcaata ttttagtagc tcgttacagt
9901 ccggtgcgtt tttggttttt tgaaagtgcg tcatcagagc gcttttggtt ttcaaaagcg
9961 ctctgaagtt cctatacttt ctagctagag aataggaact tcccgagcgg ccgcgtgtta
10021 caaccaatta accaattctg attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
10081 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
10141 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
10201 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
10261 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagcttat gcatttcttt
10321 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
10381 accgttattc attcgtgatt gcgcctgagc gaggcgaaat acgcgatcgc tgttaaaagg
10441 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
10501 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
10561 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
10621 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
10681 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
10741 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
10801 catgttggaa tttaatcgcg gcctggagca agacgtttcc cgttgaatat ggctcataac
10861 accccttgta ttactgttta tgtaagcaga cagttttatt gttcatgatg atatattttt
10921 atcttgtgca atgtaacatc agagattttg agacacaacg tggctttgtt gaataaatcg
10981 aacttttgct gagttgaagg atcagtcatg accaaaatcc cttaacgtga gttttcgttc
11041 cactgagcgt cagaccccgt agaaaagatc aaaggatctt c
//
