| 质粒名称: | pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101217 |
| 存储实验室: | Jennifer Doudna, Keith Joung |
|
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 K848A/K1003A/R1060A/N497A/R661A/Q695A/Q926A |
| 物种来源: | 化脓性链球菌 |
| 插入大小: | -- |
| 基因突变: | K848A, K1003A, R1060A, N497A, R661A, Q695A and Q926A |
| 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: | pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant was a gift from Jennifer Doudna & Keith Joung (Addgene plasmid # 101217 ; http://n2t.net/addgene:101217 ; RRID:Addgene_101217) |
| 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 |
pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.
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 pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant 10073 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant.
ACCESSION .
VERSION .
KEYWORDS pJSC270 - Bacterial expression plasmid for SpCas9 eSpCas9(1.1)-HF1 variant
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 12, 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 393..412
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 496..515
/label=T7
/note="T7 promoter, forward primer"
promoter 496..514
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 603..625
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 645..674
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 684..1784
/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 1758..1781
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 1839..1859
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 1866..5972
/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
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLADDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRAITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPALESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKAPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
primer_bind complement(6098..6116)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 6112..6159
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6525..6553)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(6580..6598)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6666..6770
/gene="bla"
/label=AmpR promoter
CDS 6771..7631
/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(6989..7008)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7802..8390
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8291..8310
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8544..8561
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 8576..8715
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(8641..8660)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8801..8823
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(8817..9008)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 aataccgcaa gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa
61 atgacccaga gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata
121 agtgcggcga cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct
181 ctcaagggca tcggtcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt
241 agtaggttga ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg
301 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
361 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
421 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc
481 tcgatcccgc gaaattaata cgactcacta tagggagacc acaacggttt ccctctagtg
541 ccggctccgg agagctcttt aattaagcgg ccgccctgca ggactcgagt tctagaaata
601 attttgttta actttaagaa ggagatatac atatgaaatc ttctcaccat caccatcacc
661 atcaccatca ccatggttct tctatgaaaa tcgaagaagg taaactggta atctggatta
721 acggcgataa aggctataac ggtctcgctg aagtcggtaa gaaattcgag aaagataccg
781 gaattaaagt caccgttgag catccggata aactggaaga gaaattccca caggttgcgg
841 caactggcga tggccctgac attatcttct gggcacacga ccgctttggt ggctacgctc
901 aatctggcct gttggctgaa atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt
961 ttacctggga tgccgtacgt tacaacggca agctgattgc ttacccgatc gctgttgaag
1021 cgttatcgct gatttataac aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga
1081 tcccggcgct ggataaagaa ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc
1141 aagaaccgta cttcacctgg ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg
1201 aaaacggcaa gtacgacatt aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc
1261 tgaccttcct ggttgacctg attaaaaaca aacacatgaa tgcagacacc gattactcca
1321 tcgcagaagc tgcctttaat aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat
1381 ggtccaacat cgacaccagc aaagtgaatt atggtgtaac ggtactgccg accttcaagg
1441 gtcaaccatc caaaccgttc gttggcgtgc tgagcgcagg tattaacgcc gccagtccga
1501 acaaagagct ggcaaaagag ttcctcgaaa actatctgct gactgatgaa ggtctggaag
1561 cggttaataa agacaaaccg ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg
1621 cgaaagatcc acgtattgcc gccactatgg aaaacgccca gaaaggtgaa atcatgccga
1681 acatcccgca gatgtccgct ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca
1741 gcggtcgtca gactgtcgat gaagccctga aagacgcgca gactaattcg agctcgaaca
1801 acaacaacaa taacaataac aacaacctcg ggatcgagga aaacctgtac ttccaatcca
1861 atgcaatgga taagaaatac tcaataggct tagatatcgg cacaaatagc gtcggatggg
1921 cggtgatcac tgatgaatat aaggttccgt ctaaaaagtt caaggttctg ggaaatacag
1981 accgccacag tatcaaaaaa aatcttatag gggctctttt atttgacagt ggagagacag
2041 cggaagcgac tcgtctcaaa cggacagctc gtagaaggta tacacgtcgg aagaatcgta
2101 tttgttatct acaggagatt ttttcaaatg agatggcgaa agtagatgat agtttctttc
2161 atcgacttga agagtctttt ttggtggaag aagacaagaa gcatgaacgt catcctattt
2221 ttggaaatat agtagatgaa gttgcttatc atgagaaata tccaactatc tatcatctgc
2281 gaaaaaaatt ggtagattct actgataaag cggatttgcg cttaatctat ttggccttag
2341 cgcatatgat taagtttcgt ggtcattttt tgattgaggg agatttaaat cctgataata
2401 gtgatgtgga caaactattt atccagttgg tacaaaccta caatcaatta tttgaagaaa
2461 accctattaa cgcaagtgga gtagatgcta aagcgattct ttctgcacga ttgagtaaat
2521 caagacgatt agaaaatctc attgctcagc tccccggtga gaagaaaaat ggcttatttg
2581 ggaatctcat tgctttgtca ttgggtttga cccctaattt taaatcaaat tttgatttgg
2641 cagaagatgc taaattacag ctttcaaaag atacttacga tgatgattta gataatttat
2701 tggcgcaaat tggagatcaa tatgctgatt tgtttttggc agctaagaat ttatcagatg
2761 ctattttact ttcagatatc ctaagagtaa atactgaaat aactaaggct cccctatcag
2821 cttcaatgat taaacgctac gatgaacatc atcaagactt gactctttta aaagctttag
2881 ttcgacaaca acttccagaa aagtataaag aaatcttttt tgatcaatca aaaaacggat
2941 atgcaggtta tattgatggg ggagctagcc aagaagaatt ttataaattt atcaaaccaa
3001 ttttagaaaa aatggatggt actgaggaat tattggtgaa actaaatcgt gaagatttgc
3061 tgcgcaagca acggaccttt gacaacggct ctattcccca tcaaattcac ttgggtgagc
3121 tgcatgctat tttgagaaga caagaagact tttatccatt tttaaaagac aatcgtgaga
3181 agattgaaaa aatcttgact tttcgaattc cttattatgt tggtccattg gcgcgtggca
3241 atagtcgttt tgcatggatg actcggaagt ctgaagaaac aattacccca tggaattttg
3301 aagaagttgt cgataaaggt gcttcagctc aatcatttat tgaacgcatg acagcgtttg
3361 ataaaaatct tccaaatgaa aaagtactac caaaacatag tttgctttat gagtatttta
3421 cggtttataa cgaattgaca aaggtcaaat atgttactga aggaatgcga aaaccagcat
3481 ttctttcagg tgaacagaag aaagccattg ttgatttact cttcaaaaca aatcgaaaag
3541 taaccgttaa gcaattaaaa gaagattatt tcaaaaaaat agaatgtttt gatagtgttg
3601 aaatttcagg agttgaagat agatttaatg cttcattagg tacctaccat gatttgctaa
3661 aaattattaa agataaagat tttttggata atgaagaaaa tgaagatatc ttagaggata
3721 ttgttttaac attgacctta tttgaagata gggagatgat tgaggaaaga cttaaaacat
3781 atgctcacct ctttgatgat aaggtgatga aacagcttaa acgtcgccgt tatactggtt
3841 ggggagcgtt gtctcgaaaa ttgattaatg gtattaggga taagcaatct ggcaaaacaa
3901 tattagattt tttgaaatca gatggttttg ccaatcgcaa ttttatggcc ctgatccatg
3961 atgatagttt gacatttaaa gaagacattc aaaaagcaca agtgtctgga caaggcgata
4021 gtttacatga acatattgca aatttagctg gtagccctgc tattaaaaaa ggtattttac
4081 agactgtaaa agttgttgat gaattggtca aagtaatggg gcggcataag ccagaaaata
4141 tcgttattga aatggcacgt gaaaatcaga caactcaaaa gggccagaaa aattcgcgag
4201 agcgtatgaa acgaatcgaa gaaggtatca aagaattagg aagtcagatt cttaaagagc
4261 atcctgttga aaatactcaa ttgcaaaatg aaaagctcta tctctattat ctccaaaatg
4321 gaagagacat gtatgtggac caagaattag atattaatcg tttaagtgat tatgatgtcg
4381 atcacattgt tccacaaagt ttccttgccg acgattcaat agacaataag gtcttaacgc
4441 gttctgataa aaatcgtggt aaatcggata acgttccaag tgaagaagta gtcaaaaaga
4501 tgaaaaacta ttggagacaa cttctaaacg ccaagttaat cactcaacgt aagtttgata
4561 atttaacgaa agctgaacgt ggaggtttga gtgaacttga taaagctggt tttatcaaac
4621 gccaattggt tgaaactcgc gccatcacta agcatgtggc acaaattttg gatagtcgca
4681 tgaatactaa atacgatgaa aatgataaac ttattcgaga ggttaaagtg attaccttaa
4741 aatctaaatt agtttctgac ttccgaaaag atttccaatt ctataaagta cgtgagatta
4801 acaattacca tcatgcccat gatgcgtatc taaatgccgt cgttggaact gctttgatta
4861 agaaatatcc agcgcttgaa tcggagtttg tctatggtga ttataaagtt tatgatgttc
4921 gtaaaatgat tgctaagtct gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt
4981 actctaatat catgaacttc ttcaaaacag aaattacact tgcaaatgga gagattcgca
5041 aagcccctct aatcgaaact aatggggaaa ctggagaaat tgtctgggat aaagggcgag
5101 attttgccac agtgcgcaaa gtattgtcca tgccccaagt caatattgtc aagaaaacag
5161 aagtacagac aggcggattc tccaaggagt caattttacc aaaaagaaat tcggacaagc
5221 ttattgctcg taaaaaagac tgggatccaa aaaaatatgg tggttttgat agtccaacgg
5281 tagcttattc agtcctagtg gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat
5341 ccgttaaaga gttactaggg atcacaatta tggaaagaag ttcctttgaa aaaaatccga
5401 ttgacttttt agaagctaaa ggatataagg aagttaaaaa agacttaatc attaaactac
5461 ctaaatatag tctttttgag ttagaaaacg gtcgtaaacg gatgctggct agtgccggag
5521 aattacaaaa aggaaatgag ctggctctgc caagcaaata tgtgaatttt ttatatttag
5581 ctagtcatta tgaaaagttg aagggtagtc cagaagataa cgaacaaaaa caattgtttg
5641 tggagcagca taagcattat ttagatgaga ttattgagca aatcagtgaa ttttctaagc
5701 gtgttatttt agcagatgcc aatttagata aagttcttag tgcatataac aaacatagag
5761 acaaaccaat acgtgaacaa gcagaaaata ttattcattt atttacgttg acgaatcttg
5821 gagctcccgc tgcttttaaa tattttgata caacaattga tcgtaaacga tatacgtcta
5881 caaaagaagt tttagatgcc actcttatcc atcaatccat cactggtctt tatgaaacac
5941 gcattgattt gagtcagcta ggaggtgact aataacattg gaagtggata acggatccgc
6001 gatcgcggcg cgccacctgg tggccggccg gtaccacgcg tgcgcgctga tccggctgct
6061 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
6121 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
6181 ggatatccac aggacgggtg tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag
6241 cgaagcgagc aggactgggc ggcggccaaa gcggtcggac agtgctccga gaacgggtgc
6301 gcatagaaat tgcatcaacg catatagcgc tagcagcacg ccatagtgac tggcgatgct
6361 gtcggaatgg acgatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc
6421 tacagcatcc agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca
6481 cggtgcctga ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat
6541 gataagctgt caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt
6601 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
6661 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
6721 tgagacaata accctgataa atgcttcaat aacattgaaa aaggaagagt atgagtattc
6781 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
6841 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
6901 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
6961 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg
7021 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
7081 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
7141 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
7201 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
7261 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa
7321 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
7381 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
7441 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
7501 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
7561 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
7621 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
7681 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
7741 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
7801 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
7861 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
7921 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
7981 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
8041 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
8101 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
8161 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
8221 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
8281 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
8341 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
8401 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
8461 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
8521 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
8581 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
8641 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
8701 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
8761 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
8821 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
8881 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
8941 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
9001 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
9061 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
9121 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
9181 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
9241 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
9301 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
9361 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
9421 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag
9481 gacccaacgc tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat
9541 atgttctgcc aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca
9601 attcttggag tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg
9661 cccggctcca tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta
9721 caatccatgc caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca
9781 gcggtccaat gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct
9841 gatggtcgtc atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc
9901 cggaagcgag aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca
9961 agacgtagcc cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac
10021 gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccg
//
