| 质粒名称: | Fuw-dCas9-Dnmt3a |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 84476 |
| 存储实验室: | Rudolf Jaenisch |
|
载体骨架基本信息 |
|
| 载体名称: | FUW |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 9207 bp |
| 完整质粒大小: | 16149 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达,慢病毒 |
| 筛选标记: | Zeocin；抗性标记在LTRs之外，不会被包装成病毒 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9-Dnmt3a |
| 物种来源: | H. sapiens (human), Synthetic |
| 插入大小: | 6933 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AscI(未破坏) |
| 3' 克隆位点: | EcoRI (未破坏) |
| 5' 测序引物 | aactatgcgctcggg |
| 3' 测序引物 | taaagcagcgtatcc |
|
文献引用方法 |
|
| 材料和方法部分: | Fuw-dCas9-Dnmt3a was a gift from Rudolf Jaenisch (Addgene plasmid # 84476 ; http://n2t.net/addgene:84476 ; RRID:Addgene_84476) |
| 参考文献部分: | Editing DNA Methylation in the Mammalian Genome. Liu XS, Wu H, Ji X, Stelzer Y, Wu X, Czauderna S, Shu J, Dadon D, Young RA, Jaenisch R. Cell. 2016 Sep 22;167(1):233-247.e17. doi: 10.1016/j.cell.2016.08.056. 10.1016/j.cell.2016.08.056 PubMed 27662091 |
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 Fuw-dCas9-Dnmt3a 16074 bp ds-DNA circular SYN 20-AUG-2019
DEFINITION Lentiviral construct to express dCas9 fused with Dnmt3a.
ACCESSION .
VERSION .
KEYWORDS Fuw-dCas9-Dnmt3a
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16074)
AUTHORS Liu XS, Wu H, Ji X, Stelzer Y, Wu X, Czauderna S, Shu J, Dadon D,
Young RA, Jaenisch R
TITLE Editing DNA Methylation in the Mammalian Genome.
JOURNAL Cell. 2016 Sep 22;167(1):233-247.e17. doi:
10.1016/j.cell.2016.08.056.
PUBMED 27662091
REFERENCE 2 (bases 1 to 16074)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 20, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..16074
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 209..230
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(349..369)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 393..603
/label=SV40 promoter
/note="SV40 early promoter"
promoter 415..611
/label=SV40 promoter
/note="SV40 early promoter"
rep_origin 462..597
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 524..543
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 659..706
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 725..1099
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 1229..1350
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1266..1285)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 1320..1339
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(1399..1415)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(1399..1415)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(1412..1434)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 1423..1439
/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(1447..1477)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 1492..1513
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(1630..1647)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(1801..2389)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(1881..1900)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(2560..3420)
/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 3183..3202
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(3421..3525)
/gene="bla"
/label=AmpR promoter
primer_bind complement(3600..3619)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 3791..4170
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 4172..4370
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 4325..4345
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 4388..4568
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 4615..4740
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5233..5466
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 5993..6110
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6175..7386
/label=UbC promoter
/note="human ubiquitin C promoter"
primer_bind 7223..7242
/label=hUBCpro-F
/note="Human Ubiquitin C (UbC) promoter, forward primer"
CDS 7437..11540
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 11553..11573
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 11577..11597
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 11601..11621
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 11628..14366
/codon_start=1
/product="DNA methyltransferase 3A"
/label=DNMT3A
/translation="MPAMPSSGPGDTSSSAAEREEDRKDGEEQEEPRGKEERQEPSTTA
RKVGRPGRKRKHPPVESGDTPKDPAVISKSPSMAQDSGASELLPNGDLEKRSEPQPEEG
SPAGGQKGGAPAEGEGAAETLPEASRAVENGCCTPKEGRGAPAEAGKEQKETNIESMKM
EGSRGRLRGGLGWESSLRQRPMPRLTFQAGDPYYISKRKRDEWLARWKREAEKKAKVIA
GMNAVEENQGPGESQKVEEASPPAVQQPTDPASPTVATTPEPVGSDAGDKNATKAGDDE
PEYEDGRGFGIGELVWGKLRGFSWWPGRIVSWWMTGRSRAAEGTRWVMWFGDGKFSVVC
VEKLMPLSSFCSAFHQATYNKQPMYRKAIYEVLQVASSRAGKLFPVCHDSDESDTAKAV
EVQNKPMIEWALGGFQPSGPKGLEPPEEEKNPYKEVYTDMWVEPEAAAYAPPPPAKKPR
KSTAEKPKVKEIIDERTRERLVYEVRQKCRNIEDICISCGSLNVTLEHPLFVGGMCQNC
KNCFLECAYQYDDDGYQSYCTICCGGREVLMCGNNNCCRCFCVECVDLLVGPGAAQAAI
KEDPWNCYMCGHKGTYGLLRRREDWPSRLQMFFANNHDQEFDPPKVYPPVPAEKRKPIR
VLSLFDGIATGLLVLKDLGIQVDRYIASEVCEDSITVGMVRHQGKIMYVGDVRSVTQKH
IQEWGPFDLVIGGSPCNDLSIVNPARKGLYEGTGRLFFEFYRLLHDARPKEGDDRPFFW
LFENVVAMGVSDKRDISRFLESNPVMIDAKEVSAAHRARYFWGNLPGMNRPLASTVNDK
LELQECLEHGRIAKFSKVRTITTRSNSIKQGKDQHFPVFMNEKEDILWCTEMERVFGFP
VHYTDVSNMSRLARQRLLGRSWSVPVIRHLFAPLKEYFACV"
misc_feature 14391..14979
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(14444..14464)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(14862..14873)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(14982..14998)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(14983..14999)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 15504..15684
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(15710..15727)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 15716..15940
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 15986..340
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(16073..18)
/label=F1ori-R
/note="F1 origin, reverse primer"
ORIGIN
1 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
61 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
121 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
181 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
241 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
301 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
361 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
421 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
481 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
541 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
601 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt
661 tgacaattaa tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta
721 aaccatggcc aagttgacca gtgccgttcc ggtgctcacc gcgcgcgacg tcgccggagc
781 ggtcgagttc tggaccgacc ggctcgggtt ctcccgggac ttcgtggagg acgacttcgc
841 cggtgtggtc cgggacgacg tgaccctgtt catcagcgcg gtccaggacc aggtggtgcc
901 ggacaacacc ctggcctggg tgtgggtgcg cggcctggac gagctgtacg ccgagtggtc
961 ggaggtcgtg tccacgaact tccgggacgc ctccgggccg gccatgaccg agatcggcga
1021 gcagccgtgg gggcgggagt tcgccctgcg cgacccggcc ggcaactgcg tgcacttcgt
1081 ggccgaggag caggactgac acgtgctacg agatttcgat tccaccgccg ccttctatga
1141 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
1201 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
1261 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
1321 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
1381 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
1441 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
1501 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
1561 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
1621 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
1681 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
1741 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
1801 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
1861 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
1921 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
1981 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
2041 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
2101 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
2161 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
2221 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
2281 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
2341 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
2401 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
2461 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
2521 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
2581 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
2641 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
2701 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
2761 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
2821 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
2881 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
2941 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
3001 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
3061 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
3121 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
3181 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
3241 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
3301 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
3361 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
3421 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
3481 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
3541 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
3601 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
3661 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
3721 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
3781 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
3841 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
3901 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
3961 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
4021 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
4081 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
4141 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
4201 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
4261 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
4321 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag cgcgttttgc
4381 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
4441 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
4501 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
4561 ctctagcagt ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct
4621 cgacgcagga ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg
4681 agtacgccaa aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc
4741 agtattaagc gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga
4801 aagaaaaaat ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca
4861 gttaatcctg gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa
4921 ccatcccttc agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc
4981 tattgtgtgc atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag
5041 gaagagcaaa acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg
5101 aggaggagat atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat
5161 tgaaccatta ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag
5221 agcagtggga ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg
5281 cgcagcgtca atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca
5341 gcagaacaat ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg
5401 gggcatcaag cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca
5461 gctcctgggg atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa
5521 tgctagttgg agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg
5581 ggacagagaa attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa
5641 ccagcaagaa aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa
5701 ttggtttaac ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg
5761 cttggtaggt ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg
5821 atattcacca ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga
5881 aggaatagaa gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg
5941 atcggcactg cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa
6001 gaaaaggggg gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag
6061 acatacaaac taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt
6121 acagggacag cagagatcca gtttggttaa ttaacccgtg tcggctccag atctggcctc
6181 cgcgccgggt tttggcgcct cccgcgggcg cccccctcct cacggcgagc gctgccacgt
6241 cagacgaagg gcgcagcgag cgtcctgatc cttccgcccg gacgctcagg acagcggccc
6301 gctgctcata agactcggcc ttagaacccc agtatcagca gaaggacatt ttaggacggg
6361 acttgggtga ctctagggca ctggttttct ttccagagag cggaacaggc gaggaaaagt
6421 agtcccttct cggcgattct gcggagggat ctccgtgggg cggtgaacgc cgatgattat
6481 ataaggacgc gccgggtgtg gcacagctag ttccgtcgca gccgggattt gggtcgcggt
6541 tcttgtttgt ggatcgctgt gatcgtcact tggtgagtag cgggctgctg ggctggccgg
6601 ggctttcgtg gccgccgggc cgctcggtgg gacggaagcg tgtggagaga ccgccaaggg
6661 ctgtagtctg ggtccgcgag caaggttgcc ctgaactggg ggttgggggg agcgcagcaa
6721 aatggcggct gttcccgagt cttgaatgga agacgcttgt gaggcgggct gtgaggtcgt
6781 tgaaacaagg tggggggcat ggtgggcggc aagaacccaa ggtcttgagg ccttcgctaa
6841 tgcgggaaag ctcttattcg ggtgagatgg gctggggcac catctgggga ccctgacgtg
6901 aagtttgtca ctgactggag aactcggttt gtcgtctgtt gcgggggcgg cagttatggc
6961 ggtgccgttg ggcagtgcac ccgtaccttt gggagcgcgc gccctcgtcg tgtcgtgacg
7021 tcacccgttc tgttggctta taatgcaggg tggggccacc tgccggtagg tgtgcggtag
7081 gcttttctcc gtcgcaggac gcagggttcg ggcctagggt aggctctcct gaatcgacag
7141 gcgccggacc tctggtgagg ggagggataa gtgaggcgtc agtttctttg gtcggtttta
7201 tgtacctatc ttcttaagta gctgaagctc cggttttgaa ctatgcgctc ggggttggcg
7261 agtgtgtttt gtgaagtttt ttaggcacct tttgaaatgt aatcatttgg gtcaatatgt
7321 aattttcagt gttagactag taaattgtcc gctaaattct ggccgttttt ggcttttttg
7381 ttagacgaag cttgggctgc aggtcgactc tagaagttaa cctcgagggc gcgcccatgg
7441 acaagaagta ttctatcgga ctggccatcg ggactaatag cgtcgggtgg gccgtgatca
7501 ctgacgagta caaggtgccc tctaagaagt tcaaggtgct cgggaacacc gaccggcatt
7561 ccatcaagaa aaatctgatc ggagctctcc tctttgattc aggggagacc gctgaagcaa
7621 cccgcctcaa gcggactgct agacggcggt acaccaggag gaagaaccgg atttgttacc
7681 ttcaagagat attctccaac gaaatggcaa aggtcgacga cagcttcttc cataggctgg
7741 aagaatcatt cctcgtggaa gaggataaga agcatgaacg gcatcccatc ttcggtaata
7801 tcgtcgacga ggtggcctat cacgagaaat acccaaccat ctaccatctt cgcaaaaagc
7861 tggtggactc aaccgacaag gcagacctcc ggcttatcta cctggccctg gcccacatga
7921 tcaagttcag aggccacttc ctgatcgagg gcgacctcaa tcctgacaat agcgatgtgg
7981 ataaactgtt catccagctg gtgcagactt acaaccagct ctttgaagag aaccccatca
8041 atgcaagcgg agtcgatgcc aaggccattc tgtcagcccg gctgtcaaag agccgcagac
8101 ttgagaatct tatcgctcag ctgccgggtg aaaagaaaaa tggactgttc gggaacctga
8161 ttgctctttc acttgggctg actcccaatt tcaagtctaa tttcgacctg gcagaggatg
8221 ccaagctgca actgtccaag gacacctatg atgacgatct cgacaacctc ctggcccaga
8281 tcggtgacca atacgccgac cttttccttg ctgctaagaa tctttctgac gccatcctgc
8341 tgtctgacat tctccgcgtg aacactgaaa tcaccaaggc ccctctttca gcttcaatga
8401 ttaagcggta tgatgagcac caccaggacc tgaccctgct taaggcactc gtccggcagc
8461 agcttccgga gaagtacaag gaaatcttct ttgaccagtc aaagaatgga tacgccggct
8521 acatcgacgg aggtgcctcc caagaggaat tttataagtt tatcaaacct atccttgaga
8581 agatggacgg caccgaagag ctcctcgtga aactgaatcg ggaggatctg ctgcggaagc
8641 agcgcacttt cgacaatggg agcattcccc accagatcca tcttggggag cttcacgcca
8701 tccttcggcg ccaagaggac ttctacccct ttcttaagga caacagggag aagattgaga
8761 aaattctcac tttccgcatc ccctactacg tgggacccct cgccagagga aatagccggt
8821 ttgcttggat gaccagaaag tcagaagaaa ctatcactcc ctggaacttc gaagaggtgg
8881 tggacaaggg agccagcgct cagtcattca tcgaacggat gactaacttc gataagaacc
8941 tccccaatga gaaggtcctg ccgaaacatt ccctgctcta cgagtacttt accgtgtaca
9001 acgagctgac caaggtgaaa tatgtcaccg aagggatgag gaagcccgca ttcctgtcag
9061 gcgaacaaaa gaaggcaatt gtggaccttc tgttcaagac caatagaaag gtgaccgtga
9121 agcagctgaa ggaggactat ttcaagaaaa ttgaatgctt cgactctgtg gagattagcg
9181 gggtcgaaga tcggttcaac gcaagcctgg gtacctacca tgatctgctt aagatcatca
9241 aggacaagga ttttctggac aatgaggaga acgaggacat ccttgaggac attgtcctga
9301 ctctcactct gttcgaggac cgggaaatga tcgaggagag gcttaagacc tacgcccatc
9361 tgttcgacga taaagtgatg aagcaactta aacggagaag atataccgga tggggacgcc
9421 ttagccgcaa actcatcaac ggaatccggg acaaacagag cggaaagacc attcttgatt
9481 tccttaagag cgacggattc gctaatcgca acttcatgca acttatccat gatgattccc
9541 tgacctttaa ggaggacatc cagaaggccc aagtgtctgg acaaggtgac tcactgcacg
9601 agcatatcgc aaatctggct ggttcacccg ctattaagaa gggtattctc cagaccgtga
9661 aagtcgtgga cgagctggtc aaggtgatgg gtcgccataa accagagaac attgtcatcg
9721 agatggccag ggaaaaccag actacccaga agggacagaa gaacagcagg gagcggatga
9781 aaagaattga ggaagggatt aaggagctcg ggtcacagat ccttaaagag cacccggtgg
9841 aaaacaccca gcttcagaat gagaagctct atctgtacta ccttcaaaat ggacgcgata
9901 tgtatgtgga ccaagagctt gatatcaaca ggctctcaga ctacgacgtg gacgccatcg
9961 tccctcagag cttcctcaaa gacgactcaa ttgacaataa ggtgctgact cgctcagaca
10021 agaaccgggg aaagtcagat aacgtgccct cagaggaagt cgtgaaaaag atgaagaact
10081 attggcgcca gcttctgaac gcaaagctga tcactcagcg gaagttcgac aatctcacta
10141 aggctgagag gggcggactg agcgaactgg acaaagcagg attcattaaa cggcaacttg
10201 tggagactcg gcagattact aaacatgtcg cccaaatcct tgactcacgc atgaatacca
10261 agtacgacga aaacgacaaa cttatccgcg aggtgaaggt gattaccctg aagtccaagc
10321 tggtcagcga tttcagaaag gactttcaat tctacaaagt gcgggagatc aataactatc
10381 atcatgctca tgacgcatat ctgaatgccg tggtgggaac cgccctgatc aagaagtacc
10441 caaagctgga aagcgagttc gtgtacggag actacaaggt ctacgacgtg cgcaagatga
10501 ttgccaaatc tgagcaggag atcggaaagg ccaccgcaaa gtacttcttc tacagcaaca
10561 tcatgaattt cttcaagacc gaaatcaccc ttgcaaacgg tgagatccgg aagaggccgc
10621 tcatcgagac taatggggag actggcgaaa tcgtgtggga caagggcaga gatttcgcta
10681 ccgtgcgcaa agtgctttct atgcctcaag tgaacatcgt gaagaaaacc gaggtgcaaa
10741 ccggaggctt ttctaaggaa tcaatcctcc ccaagcgcaa ctccgacaag ctcattgcaa
10801 ggaagaagga ttgggaccct aagaagtacg gcggattcga ttcaccaact gtggcttatt
10861 ctgtcctggt cgtggctaag gtggaaaaag gaaagtctaa gaagctcaag agcgtgaagg
10921 aactgctggg tatcaccatt atggagcgca gctccttcga gaagaaccca attgactttc
10981 tcgaagccaa aggttacaag gaagtcaaga aggaccttat catcaagctc ccaaagtata
11041 gcctgttcga actggagaat gggcggaagc ggatgctcgc ctccgctggc gaacttcaga
11101 agggtaatga gctggctctc ccctccaagt acgtgaattt cctctacctt gcaagccatt
11161 acgagaagct gaaggggagc cccgaggaca acgagcaaaa gcaactgttt gtggagcagc
11221 ataagcatta tctggacgag atcattgagc agatttccga gttttctaaa cgcgtcattc
11281 tcgctgatgc caacctcgat aaagtcctta gcgcatacaa taagcacaga gacaaaccaa
11341 ttcgggagca ggctgagaat atcatccacc tgttcaccct caccaatctt ggtgcccctg
11401 ccgcattcaa gtacttcgac accaccatcg accggaaacg ctatacctcc accaaagaag
11461 tgctggacgc caccctcatc caccagagca tcaccggact ttacgaaact cggattgacc
11521 tctcacagct cggaggggat gagggagctg atccaaaaaa gaagagaaag gtagacccaa
11581 agaaaaaacg aaaagtagat ccgaaaaaga agaggaaggt gggatccatg cccgccatgc
11641 cctccagcgg ccccggggac accagcagct ctgctgcgga gcgggaggag gaccgaaagg
11701 acggagagga gcaggaggag ccgcgtggca aggaggagcg ccaagagccc agcaccacgg
11761 cacggaaggt ggggcggcct gggaggaagc gcaagcaccc cccggtggaa agcggtgaca
11821 cgccaaagga ccctgcggtg atctccaagt ccccatccat ggcccaggac tcaggcgcct
11881 cagagctatt acccaatggg gacttggaga agcggagtga gccccagcca gaggagggga
11941 gccctgctgg ggggcagaag ggcggggccc cagcagaggg agagggtgca gctgagaccc
12001 tgcctgaagc ctcaagagca gtggaaaatg gctgctgcac ccccaaggag ggccgaggag
12061 cccctgcaga agcgggcaaa gaacagaagg agaccaacat cgaatccatg aaaatggagg
12121 gctcccgggg ccggctgcgg ggtggcttgg gctgggagtc cagcctccgt cagcggccca
12181 tgccgaggct caccttccag gcgggggacc cctactacat cagcaagcgc aagcgggacg
12241 agtggctggc acgctggaaa agggaggctg agaagaaagc caaggtcatt gcaggaatga
12301 atgctgtgga agaaaaccag gggcccgggg agtctcagaa ggtggaggag gccagccctc
12361 ctgctgtgca gcagcccact gaccccgcat cccccactgt ggctaccacg cctgagcccg
12421 tggggtccga tgctggggac aagaatgcca ccaaagcagg cgatgacgag ccagagtacg
12481 aggacggccg gggctttggc attggggagc tggtgtgggg gaaactgcgg ggcttctcct
12541 ggtggccagg ccgcattgtg tcttggtgga tgacgggccg gagccgagca gctgaaggca
12601 cccgctgggt catgtggttc ggagacggca aattctcagt ggtgtgtgtt gagaagctga
12661 tgccgctgag ctcgttttgc agtgcgttcc accaggccac gtacaacaag cagcccatgt
12721 accgcaaagc catctacgag gtcctgcagg tggccagcag ccgcgcgggg aagctgttcc
12781 cggtgtgcca cgacagcgat gagagtgaca ctgccaaggc cgtggaggtg cagaacaagc
12841 ccatgattga atgggccctg gggggcttcc agccttctgg ccctaagggc ctggagccac
12901 cagaagaaga gaagaatccc tacaaagaag tgtacacgga catgtgggtg gaacctgagg
12961 cagctgccta cgcaccacct ccaccagcca aaaagccccg gaagagcaca gcggagaagc
13021 ccaaggtcaa ggagattatt gatgagcgca caagagagcg gctggtgtac gaggtgcggc
13081 agaagtgccg gaacattgag gacatctgca tctcctgtgg gagcctcaat gttaccctgg
13141 aacaccccct cttcgttgga ggaatgtgcc aaaactgcaa gaactgcttt ctggagtgtg
13201 cgtaccagta cgacgacgac ggctaccagt cctactgcac catctgctgt gggggccgtg
13261 aggtgctcat gtgcggaaac aacaactgct gcaggtgctt ttgcgtggag tgtgtggacc
13321 tcttggtggg gccgggggct gcccaggcag ccattaagga agacccctgg aactgctaca
13381 tgtgcgggca caagggtacc tacgggctgc tgcggcggcg agaggactgg ccctcccggc
13441 tccagatgtt cttcgctaat aaccacgacc aggaatttga ccctccaaag gtttacccac
13501 ctgtcccagc tgagaagagg aagcccatcc gggtgctgtc tctctttgat ggaatcgcta
13561 cagggctcct ggtgctgaag gacttgggca ttcaggtgga ccgctacatt gcctcggagg
13621 tgtgtgagga ctccatcacg gtgggcatgg tgcggcacca ggggaagatc atgtacgtcg
13681 gggacgtccg cagcgtcaca cagaagcata tccaggagtg gggcccattc gatctggtga
13741 ttgggggcag tccctgcaat gacctctcca tcgtcaaccc tgctcgcaag ggcctctacg
13801 agggcactgg ccggctcttc tttgagttct accgcctcct gcatgatgcg cggcccaagg
13861 agggagatga tcgccccttc ttctggctct ttgagaatgt ggtggccatg ggcgttagtg
13921 acaagaggga catctcgcga tttctcgagt ccaaccctgt gatgattgat gccaaagaag
13981 tgtcagctgc acacagggcc cgctacttct ggggtaacct tcccggtatg aacaggccgt
14041 tggcatccac tgtgaatgat aagctggagc tgcaggagtg tctggagcat ggcaggatag
14101 ccaagttcag caaagtgagg accattacta cgaggtcaaa ctccataaag cagggcaaag
14161 accagcattt tcctgtcttc atgaatgaga aagaggacat cttatggtgc actgaaatgg
14221 aaagggtatt tggtttccca gtccactata ctgacgtctc caacatgagc cgcttggcga
14281 ggcagagact gctgggccgg tcatggagcg tgccagtcat ccgccacctc ttcgctccgc
14341 tgaaggagta ttttgcgtgt gtgtaagaat tcgatatcaa gcttatcgat aatcaacctc
14401 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
14461 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
14521 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
14581 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
14641 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
14701 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
14761 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
14821 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
14881 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
14941 ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag
15001 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
15061 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
15121 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg
15181 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
15241 acacacaagg ctacttccct gattggcaga actacacacc agggccaggg atcagatatc
15301 cactgacctt tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag
15361 ccaatgaagg agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc
15421 cggagagaga agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc
15481 gagagctgca tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc
15541 tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc
15601 aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt
15661 agtcagtgtg gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg
15721 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
15781 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt
15841 aggtgtcatt ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa
15901 gacaatagca ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc
15961 agctggggct ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt
16021 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgct
//
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