| 质粒名称: | pCas9-HE |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 109400 |
| 存储实验室: | Jean-Paul Concordet |
|
载体骨架基本信息 |
|
| 载体名称: | hCas9 Addgene ID 41815 |
| 载体出品公司: | Church lab A |
| 空载体大小: | 10438 bp |
| 完整质粒大小: | 10438 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 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: | HDR-Enhancer domain of human CtIP protein |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 888 bp |
|
文献引用方法 |
|
| 材料和方法部分: | pCas9-HE was a gift from Jean-Paul Concordet (Addgene plasmid # 109400 ; http://n2t.net/addgene:109400 ; RRID:Addgene_109400) |
| 参考文献部分: | CtIP fusion to Cas9 enhances transgene integration by homology-dependent repair. Charpentier M, Khedher AHY, Menoret S, Brion A, Lamribet K, Dardillac E, Boix C, Perrouault L, Tesson L, Geny S, De Cian A, Itier JM, Anegon I, Lopez B, Giovannangeli C, Concordet JP. Nat Commun. 2018 Mar 19;9(1):1133. doi: 10.1038/s41467-018-03475-7. 10.1038/s41467-018-03475-7 PubMed 29556040 |
买家导航
pCas9-HE载体质粒基本信息
pCas9-HE质粒图谱载体图谱和pCas9-HE载体序列质粒序列多克隆位点信息
pCas9-HE质粒载体简介
pCas9-HE质粒序列载体序列
LOCUS pCas9-HE 10438 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses Cas9 fusion to HE (HDR-enhancer) domain of human CtIP
protein.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10438)
AUTHORS Charpentier M, Khedher AHY, Menoret S, Brion A, Lamribet K,
Dardillac E, Boix C, Perrouault L, Tesson L, Geny S, De Cian A,
Itier JM, Anegon I, Lopez B, Giovannangeli C, Concordet JP
TITLE CtIP fusion to Cas9 enhances transgene integration by
homology-dependent repair.
JOURNAL Nat Commun. 2018 Mar 19;9(1):1133. doi: 10.1038/s41467-018-03475-7.
PUBMED 29556040
REFERENCE 2 (bases 1 to 10438)
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: "Nat
Commun."; date: "2018-03-19"; volume: "9(1):1133. doi"; pages: "
10.1038/s41467-018-03475-7"
COMMENT Sequence Label: pCas9-HE
FEATURES Location/Qualifiers
source 1..10438
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 68..447
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 448..651
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 602..622
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 648..672
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 760..769
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 766..4869
/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 766..4869
/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 766..4869
/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 4882..4902
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(5842..5861)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 5886..5934
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6136..6564
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6223..6242)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 6433..6454
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(6573..6593)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 6578..6907
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6758..6893
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 6820..6839
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 6974..7768
/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
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7028..7047)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 7638..7657
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 7944..8065
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(7981..8000)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8035..8054
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8114..8130)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8114..8130)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8127..8149)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8138..8154
/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)."
promoter complement(join(8162..8168,8169..8186,8187..8192))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8207..8228
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8345..8362)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8516..9104)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8596..8615)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(9275..10066,10067..10135))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 9898..9917
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10136..10240)
/gene="bla"
/label=AmpR promoter
primer_bind complement(10315..10334)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
61 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tcgtttagtg
661 aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag aagacaccgg
721 gaccgatcca gcctccggac tctagaggat cgaacccttg ccaccatgga caagaagtac
781 tccattgggc tcgatatcgg cacaaacagc gtcggctggg ccgtcattac ggacgagtac
841 aaggtgccga gcaaaaaatt caaagttctg ggcaataccg atcgccacag cataaagaag
901 aacctcattg gcgccctcct gttcgactcc ggggagacgg ccgaagccac gcggctcaaa
961 agaacagcac ggcgcagata tacccgcaga aagaatcgga tctgctacct gcaggagatc
1021 tttagtaatg agatggctaa ggtggatgac tctttcttcc ataggctgga ggagtccttt
1081 ttggtggagg aggataaaaa gcacgagcgc cacccaatct ttggcaatat cgtggacgag
1141 gtggcgtacc atgaaaagta cccaaccata tatcatctga ggaagaagct tgtagacagt
1201 actgataagg ctgacttgcg gttgatctat ctcgcgctgg cgcatatgat caaatttcgg
1261 ggacacttcc tcatcgaggg ggacctgaac ccagacaaca gcgatgtcga caaactcttt
1321 atccaactgg ttcagactta caatcagctt ttcgaagaga acccgatcaa cgcatccgga
1381 gttgacgcca aagcaatcct gagcgctagg ctgtccaaat cccggcggct cgaaaacctc
1441 atcgcacagc tccctgggga gaagaagaac ggcctgtttg gtaatcttat cgccctgtca
1501 ctcgggctga cccccaactt taaatctaac ttcgacctgg ccgaagatgc caagcttcaa
1561 ctgagcaaag acacctacga tgatgatctc gacaatctgc tggcccagat cggcgaccag
1621 tacgcagacc tttttttggc ggcaaagaac ctgtcagacg ccattctgct gagtgatatt
1681 ctgcgagtga acacggagat caccaaagct ccgctgagcg ctagtatgat caagcgctat
1741 gatgagcacc accaagactt gactttgctg aaggcccttg tcagacagca actgcctgag
1801 aagtacaagg aaattttctt cgatcagtct aaaaatggct acgccggata cattgacggc
1861 ggagcaagcc aggaggaatt ttacaaattt attaagccca tcttggaaaa aatggacggc
1921 accgaggagc tgctggtaaa gcttaacaga gaagatctgt tgcgcaaaca gcgcactttc
1981 gacaatggaa gcatccccca ccagattcac ctgggcgaac tgcacgctat cctcaggcgg
2041 caagaggatt tctacccctt tttgaaagat aacagggaaa agattgagaa aatcctcaca
2101 tttcggatac cctactatgt aggccccctc gcccggggaa attccagatt cgcgtggatg
2161 actcgcaaat cagaagagac catcactccc tggaacttcg aggaagtcgt ggataagggg
2221 gcctctgccc agtccttcat cgaaaggatg actaactttg ataaaaatct gcctaacgaa
2281 aaggtgcttc ctaaacactc tctgctgtac gagtacttca cagtttataa cgagctcacc
2341 aaggtcaaat acgtcacaga agggatgaga aagccagcat tcctgtctgg agagcagaag
2401 aaagctatcg tggacctcct cttcaagacg aaccggaaag ttaccgtgaa acagctcaaa
2461 gaagactatt tcaaaaagat tgaatgtttc gactctgttg aaatcagcgg agtggaggat
2521 cgcttcaacg catccctggg aacgtatcac gatctcctga aaatcattaa agacaaggac
2581 ttcctggaca atgaggagaa cgaggacatt cttgaggaca ttgtcctcac ccttacgttg
2641 tttgaagata gggagatgat tgaagaacgc ttgaaaactt acgctcatct cttcgacgac
2701 aaagtcatga aacagctcaa gaggcgccga tatacaggat gggggcggct gtcaagaaaa
2761 ctgatcaatg ggatccgaga caagcagagt ggaaagacaa tcctggattt tcttaagtcc
2821 gatggatttg ccaaccggaa cttcatgcag ttgatccatg atgactctct cacctttaag
2881 gaggacatcc agaaagcaca agtttctggc cagggggaca gtcttcacga gcacatcgct
2941 aatcttgcag gtagcccagc tatcaaaaag ggaatactgc agaccgttaa ggtcgtggat
3001 gaactcgtca aagtaatggg aaggcataag cccgagaata tcgttatcga gatggcccga
3061 gagaaccaaa ctacccagaa gggacagaag aacagtaggg aaaggatgaa gaggattgaa
3121 gagggtataa aagaactggg gtcccaaatc cttaaggaac acccagttga aaacacccag
3181 cttcagaatg agaagctcta cctgtactac ctgcagaacg gcagggacat gtacgtggat
3241 caggaactgg acatcaatcg gctctccgac tacgacgtgg atcatatcgt gccccagtct
3301 tttctcaaag atgattctat tgataataaa gtgttgacaa gatccgataa aaatagaggg
3361 aagagtgata acgtcccctc agaagaagtt gtcaagaaaa tgaaaaatta ttggcggcag
3421 ctgctgaacg ccaaactgat cacacaacgg aagttcgata atctgactaa ggctgaacga
3481 ggtggcctgt ctgagttgga taaagccggc ttcatcaaaa ggcagcttgt tgagacacgc
3541 cagatcacca agcacgtggc ccaaattctc gattcacgca tgaacaccaa gtacgatgaa
3601 aatgacaaac tgattcgaga ggtgaaagtt attactctga agtctaagct ggtctcagat
3661 ttcagaaagg actttcagtt ttataaggtg agagagatca acaattacca ccatgcgcat
3721 gatgcctacc tgaatgcagt ggtaggcact gcacttatca aaaaatatcc caagcttgaa
3781 tctgaatttg tttacggaga ctataaagtg tacgatgtta ggaaaatgat cgcaaagtct
3841 gagcaggaaa taggcaaggc caccgctaag tacttctttt acagcaatat tatgaatttt
3901 ttcaagaccg agattacact ggccaatgga gagattcgga agcgaccact tatcgaaaca
3961 aacggagaaa caggagaaat cgtgtgggac aagggtaggg atttcgcgac agtccggaag
4021 gtcctgtcca tgccgcaggt gaacatcgtt aaaaagaccg aagtacagac cggaggcttc
4081 tccaaggaaa gtatcctccc gaaaaggaac agcgacaagc tgatcgcacg caaaaaagat
4141 tgggacccca agaaatacgg cggattcgat tctcctacag tcgcttacag tgtactggtt
4201 gtggccaaag tggagaaagg gaagtctaaa aaactcaaaa gcgtcaagga actgctgggc
4261 atcacaatca tggagcgatc aagcttcgaa aaaaacccca tcgactttct cgaggcgaaa
4321 ggatataaag aggtcaaaaa agacctcatc attaagcttc ccaagtactc tctctttgag
4381 cttgaaaacg gccggaaacg aatgctcgct agtgcgggcg agctgcagaa aggtaacgag
4441 ctggcactgc cctctaaata cgttaatttc ttgtatctgg ccagccacta tgaaaagctc
4501 aaagggtctc ccgaagataa tgagcagaag cagctgttcg tggaacaaca caaacactac
4561 cttgatgaga tcatcgagca aataagcgaa ttctccaaaa gagtgatcct cgccgacgct
4621 aacctcgata aggtgctttc tgcttacaat aagcacaggg ataagcccat cagggagcag
4681 gcagaaaaca ttatccactt gtttactctg accaacttgg gcgcgcctgc agccttcaag
4741 tacttcgaca ccaccataga cagaaagcgg tacacctcta caaaggaggt cctggacgcc
4801 acactgattc atcagtcaat tacggggctc tatgaaacaa gaatcgacct ctctcagctc
4861 ggtggagaca gcagggctga ccccaagaaa aagaggaagg tgggatccgc tagcatgaac
4921 atctcgggaa gcagctgtgg aagccctaac tctgcagata catctagtga ctttaaagat
4981 ctgtggacca aactaaaaga atgtcatgat agagaagtac aaggtttaca agtaaaagta
5041 accaagctta agcaagagcg aatcttagat gcacaaagac tagaagaatt cttcaccaaa
5101 aatcaacagc tgagggaaca gcagaaagtc cttcatgaag ccattaaagt tttagaagat
5161 cggttaagag caggcttatg tgatcgctgt gcagtaactg aagaacatat gcggaaaaaa
5221 cagcaagagt ttgaaaatat ccggcagcag aatcttaaac ttattacaga acttatgaat
5281 gaaaggaata ctctacagga agaaaataaa aagctttctg aacaactcca gcagaaaatt
5341 gagaatgatc aacagcatca agcagctgag cttgaatgtg aggaagacgt tattccagat
5401 tcaccgataa cagccttctc attttctggc gttaaccggc tacgaagaaa ggagaacccc
5461 catgtccgat acatagaaca aacacatact aaattggagc actctgtgtg tgcaaatgaa
5521 atgagaaaag tttccaagtc ttcaactcat ccacaacata atcctaatga aaatgaaatt
5581 ctagtagctg acacttatga ccaaagtcaa tctccaatgg ccaaagcaca tggaacaagc
5641 agctataccc ctgataagtc atcttttaat ttagctacag ttgttgctga aacacttgga
5701 cttggtgttc aagaagaatc tgaaactcaa ggtcccatga gcccccttgg tgatgagctc
5761 taccactgtc tggaaggaaa tcacaagaaa cagccttttg agtagaccgg ttagtaatga
5821 gtttaaacgg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
5881 tatgacggca ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac
5941 gcggggttcg gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc
6001 aatacgcccg cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag
6061 ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct
6121 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
6181 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
6241 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
6301 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
6361 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
6421 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
6481 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
6541 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
6601 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
6661 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
6721 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
6781 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
6841 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
6901 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg
6961 aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
7021 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
7081 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
7141 cctgaatgaa ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
7201 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
7261 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
7321 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
7381 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
7441 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
7501 gcgcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
7561 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
7621 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
7681 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
7741 ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc
7801 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
7861 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
7921 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
7981 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
8041 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
8101 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
8161 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
8221 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
8281 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
8341 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
8401 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
8461 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
8521 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
8581 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
8641 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
8701 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
8761 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
8821 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
8881 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
8941 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
9001 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
9061 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
9121 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
9181 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
9241 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
9301 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
9361 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
9421 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
9481 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
9541 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
9601 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
9661 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
9721 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
9781 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
9841 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
9901 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
9961 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
10021 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
10081 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
10141 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
10201 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
10261 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
10321 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
10381 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgc
//
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