| 质粒名称: | pJSC120 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variant |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 101208 |
| 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 C80S/C574S/S355C/S867C/N497A/R661A/Q695A |
| 物种来源: | 化脓性链球菌 |
| 插入大小: | -- |
| 基因突变: | C80S, C574S, S355C, S867C, 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: | pJSC120 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variant was a gift from Jennifer Doudna & Keith Joung (Addgene plasmid # 101208 ; http://n2t.net/addgene:101208 ; RRID:Addgene_101208) |
| 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 |
pJSC120 - 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 pJSC120 - Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH FRET variant 10073 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Bacterial expression plasmid for SpCas9 + N497A/R661A/Q695A, HNH
FRET variant.
ACCESSION .
VERSION .
KEYWORDS pJSC120 - 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 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 133..152
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 236..255
/label=T7
/note="T7 promoter, forward primer"
promoter 236..254
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 343..365
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 385..414
/codon_start=1
/product="10xHis affinity tag"
/label=10xHis
/translation="HHHHHHHHHH"
CDS 424..1524
/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 1498..1521
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 1579..1599
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
CDS 1606..5712
/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
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRISYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQCKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTAFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIESFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGALSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
ALIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKCDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
primer_bind complement(5838..5856)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5852..5899
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(6265..6293)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(6320..6338)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6406..6510
/gene="bla"
/label=AmpR promoter
CDS 6511..7371
/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(6729..6748)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 7542..8130
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8031..8050
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 8284..8301
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 8316..8455
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(8381..8400)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8541..8563
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(8557..8748)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
61 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
121 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
181 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
241 cgactcacta tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt
301 aattaagcgg ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa
361 ggagatatac atatgaaatc ttctcaccat caccatcacc atcaccatca ccatggttct
421 tctatgaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
481 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
541 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
601 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
661 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
721 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
781 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
841 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
901 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
961 aaagacgtgg gcgtggataa cgctggcgca aaagcgggtc tgaccttcct ggttgacctg
1021 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
1081 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
1141 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
1201 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
1261 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
1321 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
1381 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
1441 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
1501 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacaa taacaataac
1561 aacaacctcg ggatcgagga aaacctgtac ttccaatcca atgcaatgga taagaaatac
1621 tcaataggct tagatatcgg cacaaatagc gtcggatggg cggtgatcac tgatgaatat
1681 aaggttccgt ctaaaaagtt caaggttctg ggaaatacag accgccacag tatcaaaaaa
1741 aatcttatag gggctctttt atttgacagt ggagagacag cggaagcgac tcgtctcaaa
1801 cggacagctc gtagaaggta tacacgtcgg aagaatcgta tttcttatct acaggagatt
1861 ttttcaaatg agatggcgaa agtagatgat agtttctttc atcgacttga agagtctttt
1921 ttggtggaag aagacaagaa gcatgaacgt catcctattt ttggaaatat agtagatgaa
1981 gttgcttatc atgagaaata tccaactatc tatcatctgc gaaaaaaatt ggtagattct
2041 actgataaag cggatttgcg cttaatctat ttggccttag cgcatatgat taagtttcgt
2101 ggtcattttt tgattgaggg agatttaaat cctgataata gtgatgtgga caaactattt
2161 atccagttgg tacaaaccta caatcaatta tttgaagaaa accctattaa cgcaagtgga
2221 gtagatgcta aagcgattct ttctgcacga ttgagtaaat caagacgatt agaaaatctc
2281 attgctcagc tccccggtga gaagaaaaat ggcttatttg ggaatctcat tgctttgtca
2341 ttgggtttga cccctaattt taaatcaaat tttgatttgg cagaagatgc taaattacag
2401 ctttcaaaag atacttacga tgatgattta gataatttat tggcgcaaat tggagatcaa
2461 tatgctgatt tgtttttggc agctaagaat ttatcagatg ctattttact ttcagatatc
2521 ctaagagtaa atactgaaat aactaaggct cccctatcag cttcaatgat taaacgctac
2581 gatgaacatc atcaagactt gactctttta aaagctttag ttcgacaaca acttccagaa
2641 aagtataaag aaatcttttt tgatcaatgt aaaaacggat atgcaggtta tattgatggg
2701 ggagctagcc aagaagaatt ttataaattt atcaaaccaa ttttagaaaa aatggatggt
2761 actgaggaat tattggtgaa actaaatcgt gaagatttgc tgcgcaagca acggaccttt
2821 gacaacggct ctattcccca tcaaattcac ttgggtgagc tgcatgctat tttgagaaga
2881 caagaagact tttatccatt tttaaaagac aatcgtgaga agattgaaaa aatcttgact
2941 tttcgaattc cttattatgt tggtccattg gcgcgtggca atagtcgttt tgcatggatg
3001 actcggaagt ctgaagaaac aattacccca tggaattttg aagaagttgt cgataaaggt
3061 gcttcagctc aatcatttat tgaacgcatg acagcgtttg ataaaaatct tccaaatgaa
3121 aaagtactac caaaacatag tttgctttat gagtatttta cggtttataa cgaattgaca
3181 aaggtcaaat atgttactga aggaatgcga aaaccagcat ttctttcagg tgaacagaag
3241 aaagccattg ttgatttact cttcaaaaca aatcgaaaag taaccgttaa gcaattaaaa
3301 gaagattatt tcaaaaaaat agaatctttt gatagtgttg aaatttcagg agttgaagat
3361 agatttaatg cttcattagg tacctaccat gatttgctaa aaattattaa agataaagat
3421 tttttggata atgaagaaaa tgaagatatc ttagaggata ttgttttaac attgacctta
3481 tttgaagata gggagatgat tgaggaaaga cttaaaacat atgctcacct ctttgatgat
3541 aaggtgatga aacagcttaa acgtcgccgt tatactggtt ggggagcgtt gtctcgaaaa
3601 ttgattaatg gtattaggga taagcaatct ggcaaaacaa tattagattt tttgaaatca
3661 gatggttttg ccaatcgcaa ttttatggcc ctgatccatg atgatagttt gacatttaaa
3721 gaagacattc aaaaagcaca agtgtctgga caaggcgata gtttacatga acatattgca
3781 aatttagctg gtagccctgc tattaaaaaa ggtattttac agactgtaaa agttgttgat
3841 gaattggtca aagtaatggg gcggcataag ccagaaaata tcgttattga aatggcacgt
3901 gaaaatcaga caactcaaaa gggccagaaa aattcgcgag agcgtatgaa acgaatcgaa
3961 gaaggtatca aagaattagg aagtcagatt cttaaagagc atcctgttga aaatactcaa
4021 ttgcaaaatg aaaagctcta tctctattat ctccaaaatg gaagagacat gtatgtggac
4081 caagaattag atattaatcg tttaagtgat tatgatgtcg atcacattgt tccacaaagt
4141 ttccttaaag acgattcaat agacaataag gtcttaacgc gttctgataa aaatcgtggt
4201 aaatgcgata acgttccaag tgaagaagta gtcaaaaaga tgaaaaacta ttggagacaa
4261 cttctaaacg ccaagttaat cactcaacgt aagtttgata atttaacgaa agctgaacgt
4321 ggaggtttga gtgaacttga taaagctggt tttatcaaac gccaattggt tgaaactcgc
4381 caaatcacta agcatgtggc acaaattttg gatagtcgca tgaatactaa atacgatgaa
4441 aatgataaac ttattcgaga ggttaaagtg attaccttaa aatctaaatt agtttctgac
4501 ttccgaaaag atttccaatt ctataaagta cgtgagatta acaattacca tcatgcccat
4561 gatgcgtatc taaatgccgt cgttggaact gctttgatta agaaatatcc aaaacttgaa
4621 tcggagtttg tctatggtga ttataaagtt tatgatgttc gtaaaatgat tgctaagtct
4681 gagcaagaaa taggcaaagc aaccgcaaaa tatttctttt actctaatat catgaacttc
4741 ttcaaaacag aaattacact tgcaaatgga gagattcgca aacgccctct aatcgaaact
4801 aatggggaaa ctggagaaat tgtctgggat aaagggcgag attttgccac agtgcgcaaa
4861 gtattgtcca tgccccaagt caatattgtc aagaaaacag aagtacagac aggcggattc
4921 tccaaggagt caattttacc aaaaagaaat tcggacaagc ttattgctcg taaaaaagac
4981 tgggatccaa aaaaatatgg tggttttgat agtccaacgg tagcttattc agtcctagtg
5041 gttgctaagg tggaaaaagg gaaatcgaag aagttaaaat ccgttaaaga gttactaggg
5101 atcacaatta tggaaagaag ttcctttgaa aaaaatccga ttgacttttt agaagctaaa
5161 ggatataagg aagttaaaaa agacttaatc attaaactac ctaaatatag tctttttgag
5221 ttagaaaacg gtcgtaaacg gatgctggct agtgccggag aattacaaaa aggaaatgag
5281 ctggctctgc caagcaaata tgtgaatttt ttatatttag ctagtcatta tgaaaagttg
5341 aagggtagtc cagaagataa cgaacaaaaa caattgtttg tggagcagca taagcattat
5401 ttagatgaga ttattgagca aatcagtgaa ttttctaagc gtgttatttt agcagatgcc
5461 aatttagata aagttcttag tgcatataac aaacatagag acaaaccaat acgtgaacaa
5521 gcagaaaata ttattcattt atttacgttg acgaatcttg gagctcccgc tgcttttaaa
5581 tattttgata caacaattga tcgtaaacga tatacgtcta caaaagaagt tttagatgcc
5641 actcttatcc atcaatccat cactggtctt tatgaaacac gcattgattt gagtcagcta
5701 ggaggtgact aataacattg gaagtggata acggatccgc gatcgcggcg cgccacctgg
5761 tggccggccg gtaccacgcg tgcgctctga tccggctgct aacaaagccc gaaaggaagc
5821 tgagttggct gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg
5881 ggtcttgagg ggttttttgc tgaaaggagg aactatatcc ggatatccac aggacgggtg
5941 tggtcgccat gatcgcgtag tcgatagtgg ctccaagtag cgaagcgagc aggactgggc
6001 ggcggccaaa gcggtcggac agtgctccga gaacgggtgc gcatagaaat tgcatcaacg
6061 catatagcgc tagcagcacg ccatagtgac tggcgatgct gtcggaatgg acgatatccc
6121 gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc agggtgacgg
6181 tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga ctgcgttagc
6241 aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt caaacatgag
6301 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
6361 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
6421 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
6481 atgcttcaat aacattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
6541 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
6601 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
6661 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
6721 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
6781 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
6841 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
6901 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
6961 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
7021 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
7081 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
7141 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
7201 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
7261 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
7321 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
7381 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
7441 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
7501 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
7561 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
7621 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
7681 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
7741 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
7801 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
7861 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
7921 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
7981 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
8041 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
8101 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
8161 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
8221 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
8281 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
8341 catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc tctgatgccg
8401 catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg
8461 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
8521 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
8581 gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg attcacagat
8641 gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta atgtctggct
8701 tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg atgcctccgt
8761 gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga ggatgctcac
8821 gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg gtaaacaact
8881 ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc agcgcttcgt
8941 taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc agatccggaa
9001 cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac ggaaaccgaa
9061 gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc ttcacgttcg
9121 ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc ctagccgggt
9181 cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc tgcccgagat
9241 gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc aagggttggt
9301 ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag tggtgaatcc
9361 gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca tgcaccgcga
9421 cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc caacccgttc
9481 catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat gatcgaagtt
9541 aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc atctacctgc
9601 ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag aagaatcata
9661 atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc cagcgcgtcg
9721 gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg
9781 acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag gccgatcatc
9841 gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt
9901 cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt catgccccgc
9961 gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg acgctctccc
10021 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gag
//
