| 质粒名称: | pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101207 |
| 存储实验室: | Vladislav Verkhusha |
|
载体骨架基本信息 |
|
| 载体名称: | pCT10 |
| 载体出品公司: | Doudna lab |
| 空载体大小: | -- |
| 完整质粒大小: | 10073 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 筛选标记: | -- |
| 载体抗性: | 氨苄青霉素，100μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | SpCas9 variant N497A/R661A/Q695A |
| 物种来源: | 化脓性链球菌 |
| 插入大小: | -- |
| 基因突变: | N497A, R661A and Q695A |
| GenBank编号 | -- |
| 启动子: | T7 |
| 标签/融合蛋白: | 10xHis（主干上的N端子）MBP（主干上的N终端）TEV站点（主干上的N终端） |
| 克隆方法: | Gibson Cloning |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | T7 |
| 3’测序引物 | T7-term |
|
文献引用方法 |
|
| 材料和方法部分: | pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant was a gift from Jennifer Doudna & Keith Joung (Addgene plasmid # 101207 ; http://n2t.net/addgene:101207 ; RRID:Addgene_101207) |
| 参考文献部分: | Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington LB, Sternberg SH, Joung JK, Yildiz A, Doudna JA. Nature. 2017 Sep 20. doi: 10.1038/nature24268. 10.1038/nature24268 PubMed 28931002 |
pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R6
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pJSC119 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant 10073 bp ds-DNA circular SYN 13-MAY-2021
DEFINITION Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A variant.
ACCESSION .
VERSION .
KEYWORDS pJSC119 - Bacterial expression plasmid for SpCas9 +
N497A/R661...
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10073)
AUTHORS Chen JS, Dagdas YS, Kleinstiver BP, Welch MM, Sousa AA, Harrington
LB, Sternberg SH, Joung JK, Yildiz A, Doudna JA
TITLE Enhanced proofreading governs CRISPR-Cas9 targeting accuracy.
JOURNAL Nature. 2017 Sep 20. doi: 10.1038/nature24268.
PUBMED 28931002
REFERENCE 2 (bases 1 to 10073)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10073
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 86..105
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 189..208
/label=T7
/note="T7 promoter, forward primer"
promoter 189..207
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 296..318
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 338..367
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 377..1477
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 1451..1474
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 1532..1552
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 1559..5665
/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
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
primer_bind complement(5791..5809)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5805..5852
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6218..6246)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(6273..6291)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6359..6463
/gene="bla"
/label=AmpR promoter
CDS 6464..7324
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(6682..6701)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7495..8083
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7984..8003
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8237..8254
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 8269..8408
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(8334..8353)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8494..8516
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(8510..8701)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 gtcccccggc cacggggcct gccaccatac ccacgccgaa acaagcgctc atgagcccga
61 agtggcgagc ccgatcttcc ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac
121 ctgtggcgcc ggtgatgccg gccacgatgc gtccggcgta gaggatcgag atctcgatcc
181 cgcgaaatta atacgactca ctatagggag accacaacgg tttccctcta gtgccggctc
241 cggagagctc tttaattaag cggccgccct gcaggactcg agttctagaa ataattttgt
301 ttaactttaa gaaggagata tacatatgaa atcttctcac catcaccatc accatcacca
361 tcaccatggt tcttctatga aaatcgaaga aggtaaactg gtaatctgga ttaacggcga
421 taaaggctat aacggtctcg ctgaagtcgg taagaaattc gagaaagata ccggaattaa
481 agtcaccgtt gagcatccgg ataaactgga agagaaattc ccacaggttg cggcaactgg
541 cgatggccct gacattatct tctgggcaca cgaccgcttt ggtggctacg ctcaatctgg
601 cctgttggct gaaatcaccc cggacaaagc gttccaggac aagctgtatc cgtttacctg
661 ggatgccgta cgttacaacg gcaagctgat tgcttacccg atcgctgttg aagcgttatc
721 gctgatttat aacaaagatc tgctgccgaa cccgccaaaa acctgggaag agatcccggc
781 gctggataaa gaactgaaag cgaaaggtaa gagcgcgctg atgttcaacc tgcaagaacc
841 gtacttcacc tggccgctga ttgctgctga cgggggttat gcgttcaagt atgaaaacgg
901 caagtacgac attaaagacg tgggcgtgga taacgctggc gcgaaagcgg gtctgacctt
961 cctggttgac ctgattaaaa acaaacacat gaatgcagac accgattact ccatcgcaga
1021 agctgccttt aataaaggcg aaacagcgat gaccatcaac ggcccgtggg catggtccaa
1081 catcgacacc agcaaagtga attatggtgt aacggtactg ccgaccttca agggtcaacc
1141 atccaaaccg ttcgttggcg tgctgagcgc aggtattaac gccgccagtc cgaacaaaga
1201 gctggcaaaa gagttcctcg aaaactatct gctgactgat gaaggtctgg aagcggttaa
1261 taaagacaaa ccgctgggtg ccgtagcgct gaagtcttac gaggaagagt tggcgaaaga
1321 tccacgtatt gccgccacta tggaaaacgc ccagaaaggt gaaatcatgc cgaacatccc
1381 gcagatgtcc gctttctggt atgccgtgcg tactgcggtg atcaacgccg ccagcggtcg
1441 tcagactgtc gatgaagccc tgaaagacgc gcagactaat tcgagctcga acaacaacaa
1501 caataacaat aacaacaacc tcgggatcga ggaaaacctg tacttccaat ccaatgcaat
1561 ggataagaaa tactcaatag gcttagatat cggcacaaat agcgtcggat gggcggtgat
1621 cactgatgaa tataaggttc cgtctaaaaa gttcaaggtt ctgggaaata cagaccgcca
1681 cagtatcaaa aaaaatctta taggggctct tttatttgac agtggagaga cagcggaagc
1741 gactcgtctc aaacggacag ctcgtagaag gtatacacgt cggaagaatc gtatttgtta
1801 tctacaggag attttttcaa atgagatggc gaaagtagat gatagtttct ttcatcgact
1861 tgaagagtct tttttggtgg aagaagacaa gaagcatgaa cgtcatccta tttttggaaa
1921 tatagtagat gaagttgctt atcatgagaa atatccaact atctatcatc tgcgaaaaaa
1981 attggtagat tctactgata aagcggattt gcgcttaatc tatttggcct tagcgcatat
2041 gattaagttt cgtggtcatt ttttgattga gggagattta aatcctgata atagtgatgt
2101 ggacaaacta tttatccagt tggtacaaac ctacaatcaa ttatttgaag aaaaccctat
2161 taacgcaagt ggagtagatg ctaaagcgat tctttctgca cgattgagta aatcaagacg
2221 attagaaaat ctcattgctc agctccccgg tgagaagaaa aatggcttat ttgggaatct
2281 cattgctttg tcattgggtt tgacccctaa ttttaaatca aattttgatt tggcagaaga
2341 tgctaaatta cagctttcaa aagatactta cgatgatgat ttagataatt tattggcgca
2401 aattggagat caatatgctg atttgttttt ggcagctaag aatttatcag atgctatttt
2461 actttcagat atcctaagag taaatactga aataactaag gctcccctat cagcttcaat
2521 gattaaacgc tacgatgaac atcatcaaga cttgactctt ttaaaagctt tagttcgaca
2581 acaacttcca gaaaagtata aagaaatctt ttttgatcaa tcaaaaaacg gatatgcagg
2641 ttatattgat gggggagcta gccaagaaga attttataaa tttatcaaac caattttaga
2701 aaaaatggat ggtactgagg aattattggt gaaactaaat cgtgaagatt tgctgcgcaa
2761 gcaacggacc tttgacaacg gctctattcc ccatcaaatt cacttgggtg agctgcatgc
2821 tattttgaga agacaagaag acttttatcc atttttaaaa gacaatcgtg agaagattga
2881 aaaaatcttg acttttcgaa ttccttatta tgttggtcca ttggcgcgtg gcaatagtcg
2941 ttttgcatgg atgactcgga agtctgaaga aacaattacc ccatggaatt ttgaagaagt
3001 tgtcgataaa ggtgcttcag ctcaatcatt tattgaacgc atgacagcgt ttgataaaaa
3061 tcttccaaat gaaaaagtac taccaaaaca tagtttgctt tatgagtatt ttacggttta
3121 taacgaattg acaaaggtca aatatgttac tgaaggaatg cgaaaaccag catttctttc
3181 aggtgaacag aagaaagcca ttgttgattt actcttcaaa acaaatcgaa aagtaaccgt
3241 taagcaatta aaagaagatt atttcaaaaa aatagaatgt tttgatagtg ttgaaatttc
3301 aggagttgaa gatagattta atgcttcatt aggtacctac catgatttgc taaaaattat
3361 taaagataaa gattttttgg ataatgaaga aaatgaagat atcttagagg atattgtttt
3421 aacattgacc ttatttgaag atagggagat gattgaggaa agacttaaaa catatgctca
3481 cctctttgat gataaggtga tgaaacagct taaacgtcgc cgttatactg gttggggagc
3541 gttgtctcga aaattgatta atggtattag ggataagcaa tctggcaaaa caatattaga
3601 ttttttgaaa tcagatggtt ttgccaatcg caattttatg gccctgatcc atgatgatag
3661 tttgacattt aaagaagaca ttcaaaaagc acaagtgtct ggacaaggcg atagtttaca
3721 tgaacatatt gcaaatttag ctggtagccc tgctattaaa aaaggtattt tacagactgt
3781 aaaagttgtt gatgaattgg tcaaagtaat ggggcggcat aagccagaaa atatcgttat
3841 tgaaatggca cgtgaaaatc agacaactca aaagggccag aaaaattcgc gagagcgtat
3901 gaaacgaatc gaagaaggta tcaaagaatt aggaagtcag attcttaaag agcatcctgt
3961 tgaaaatact caattgcaaa atgaaaagct ctatctctat tatctccaaa atggaagaga
4021 catgtatgtg gaccaagaat tagatattaa tcgtttaagt gattatgatg tcgatcacat
4081 tgttccacaa agtttcctta aagacgattc aatagacaat aaggtcttaa cgcgttctga
4141 taaaaatcgt ggtaaatcgg ataacgttcc aagtgaagaa gtagtcaaaa agatgaaaaa
4201 ctattggaga caacttctaa acgccaagtt aatcactcaa cgtaagtttg ataatttaac
4261 gaaagctgaa cgtggaggtt tgagtgaact tgataaagct ggttttatca aacgccaatt
4321 ggttgaaact cgccaaatca ctaagcatgt ggcacaaatt ttggatagtc gcatgaatac
4381 taaatacgat gaaaatgata aacttattcg agaggttaaa gtgattacct taaaatctaa
4441 attagtttct gacttccgaa aagatttcca attctataaa gtacgtgaga ttaacaatta
4501 ccatcatgcc catgatgcgt atctaaatgc cgtcgttgga actgctttga ttaagaaata
4561 tccaaaactt gaatcggagt ttgtctatgg tgattataaa gtttatgatg ttcgtaaaat
4621 gattgctaag tctgagcaag aaataggcaa agcaaccgca aaatatttct tttactctaa
4681 tatcatgaac ttcttcaaaa cagaaattac acttgcaaat ggagagattc gcaaacgccc
4741 tctaatcgaa actaatgggg aaactggaga aattgtctgg gataaagggc gagattttgc
4801 cacagtgcgc aaagtattgt ccatgcccca agtcaatatt gtcaagaaaa cagaagtaca
4861 gacaggcgga ttctccaagg agtcaatttt accaaaaaga aattcggaca agcttattgc
4921 tcgtaaaaaa gactgggatc caaaaaaata tggtggtttt gatagtccaa cggtagctta
4981 ttcagtccta gtggttgcta aggtggaaaa agggaaatcg aagaagttaa aatccgttaa
5041 agagttacta gggatcacaa ttatggaaag aagttccttt gaaaaaaatc cgattgactt
5101 tttagaagct aaaggatata aggaagttaa aaaagactta atcattaaac tacctaaata
5161 tagtcttttt gagttagaaa acggtcgtaa acggatgctg gctagtgccg gagaattaca
5221 aaaaggaaat gagctggctc tgccaagcaa atatgtgaat tttttatatt tagctagtca
5281 ttatgaaaag ttgaagggta gtccagaaga taacgaacaa aaacaattgt ttgtggagca
5341 gcataagcat tatttagatg agattattga gcaaatcagt gaattttcta agcgtgttat
5401 tttagcagat gccaatttag ataaagttct tagtgcatat aacaaacata gagacaaacc
5461 aatacgtgaa caagcagaaa atattattca tttatttacg ttgacgaatc ttggagctcc
5521 cgctgctttt aaatattttg atacaacaat tgatcgtaaa cgatatacgt ctacaaaaga
5581 agttttagat gccactctta tccatcaatc catcactggt ctttatgaaa cacgcattga
5641 tttgagtcag ctaggaggtg actaataaca ttggaagtgg ataacggatc cgcgatcgcg
5701 gcgcgccacc tggtggccgg ccggtaccac gcgtgcgcgc tgatccggct gctaacaaag
5761 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
5821 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatatc
5881 cacaggacgg gtgtggtcgc catgatcgcg tagtcgatag tggctccaag tagcgaagcg
5941 agcaggactg ggcggcggcc aaagcggtcg gacagtgctc cgagaacggg tgcgcataga
6001 aattgcatca acgcatatag cgctagcagc acgccatagt gactggcgat gctgtcggaa
6061 tggacgatat cccgcaagag gcccggcagt accggcataa ccaagcctat gcctacagca
6121 tccagggtga cggtgccgag gatgacgatg agcgcattgt tagatttcat acacggtgcc
6181 tgactgcgtt agcaatttaa ctgtgataaa ctaccgcatt aaagcttatc gatgataagc
6241 tgtcaaacat gagaattctt gaagacgaaa gggcctcgtg atacgcctat ttttataggt
6301 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
6361 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
6421 ataaccctga taaatgcttc aataacattg aaaaaggaag agtatgagta ttcaacattt
6481 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
6541 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
6601 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
6661 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtgttg acgccgggca
6721 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
6781 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
6841 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
6901 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
6961 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgcag caatggcaac
7021 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
7081 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
7141 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
7201 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
7261 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
7321 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
7381 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
7441 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
7501 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
7561 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
7621 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
7681 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
7741 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
7801 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
7861 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
7921 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
7981 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
8041 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
8101 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
8161 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
8221 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta
8281 ttttctcctt acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc
8341 tgctctgatg ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca
8401 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
8461 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
8521 caccgtcatc accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa
8581 gcgattcaca gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg
8641 ttaatgtctg gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca
8701 ctgatgcctc cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag
8761 agaggatgct cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg
8821 agggtaaaca actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat
8881 gccagcgctt cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga
8941 tgcagatccg gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa
9001 cacggaaacc gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt
9061 cgcttcacgt tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc
9121 agcctagccg ggtcctcaac gacaggagca cgatcatgcg cacccgtggc caggacccaa
9181 cgctgcccga gatgcgccgc gtgcggctgc tggagatggc ggacgcgatg gatatgttct
9241 gccaagggtt ggtttgcgca ttcacagttc tccgcaagaa ttgattggct ccaattcttg
9301 gagtggtgaa tccgttagcg aggtgccgcc ggcttccatt caggtcgagg tggcccggct
9361 ccatgcaccg cgacgcaacg cggggaggca gacaaggtat agggcggcgc ctacaatcca
9421 tgccaacccg ttccatgtgc tcgccgaggc ggcataaatc gccgtgacga tcagcggtcc
9481 aatgatcgaa gttaggctgg taagagccgc gagcgatcct tgaagctgtc cctgatggtc
9541 gtcatctacc tgcctggaca gcatggcctg caacgcgggc atcccgatgc cgccggaagc
9601 gagaagaatc ataatgggga aggccatcca gcctcgcgtc gcgaacgcca gcaagacgta
9661 gcccagcgcg tcggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
9721 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
9781 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
9841 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
9901 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
9961 tcgacgctct cccttatgcg actcctgcat taggaagcag cccagtagta ggttgaggcc
10021 gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag atggcgccca aca
//
