| 质粒名称: | pHR-SFFV-dCas9-BFP |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 46910 |
| 存储实验室: | Stanley Qi, Jonathan Weissman |
|
载体骨架基本信息 |
|
| 载体名称: | pHR |
| 载体出品公司: | 爱迪基因 |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 9000 bp |
| 完整质粒大小: | 13936 bp |
| 载体类型: | 哺乳细胞表达，Lentiviral, CRISPR |
| 筛选标记: | 新霉素（G418） |
| 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-BFP fusion |
| 物种来源: | 人源 |
| 插入大小: | 4935 bp |
| 启动子 | SFFV |
| 标签/融合蛋白: |
HA (C terminal on insert)
2xNLS (C terminal on insert)
BFP (C terminal on insert)
|
| 克隆方法: | Ligation Independent Cloning |
| 5' 测序引物 | gcttcccgagctctataaaagag |
| 3' 测序引物 | CCAGAGGTTGATTATCGATAAGC |
|
文献引用方法 |
|
| 材料和方法部分: | pHR-SFFV-dCas9-BFP was a gift from Stanley Qi & Jonathan Weissman (爱迪基因 plasmid # 46910) |
| 参考文献部分: | CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, Weissman JS, Qi LS. Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044. 10.1016/j.cell.2013.06.044 PubMed 23849981 |
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 pHR-SFFV-dCas9-BFP 13936 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION Human expression vector containing SFFV promoter, dCas9 that is
fused to 2x NLS and tagBFP.
ACCESSION .
VERSION .
KEYWORDS pHR-SFFV-dCas9-BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13936)
AUTHORS Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE,
Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA,
Weissman JS, Qi LS
TITLE CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in
Eukaryotes.
JOURNAL Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi:
10.1016/j.cell.2013.06.044.
PUBMED 23849981
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pHR-SFFV-dCas9-BFP|
FEATURES Location/Qualifiers
source 1..13936
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1535
/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 2049..2166
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2319..2726
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 2795..2804
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2801..6904
/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 6908..6934
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 6953..6973
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6980..7000
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7040..7738
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
LPSKLGHKLN"
misc_feature 7799..8387
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7852..7872)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8270..8281)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(8390..8406)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8391..8407)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 8497..8730
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(8755..8773)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(8756..8773)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(9063..9082)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(9296..9315)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9415..9437
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(9475..9493)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 9561..9665
/gene="bla"
/label=AmpR promoter
CDS 9666..10526
/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 complement(9884..9903)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10697..11285
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 11186..11205
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 11439..11456
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(11526..11546)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 11531..11860
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 11711..11846
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 11773..11792
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 12994..13059
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 13189..13209
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 13634..13768
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(13671..13690)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 13725..13744
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
ORIGIN
1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
1501 ggctgtgaaa gatacctaaa ggatcaacag ctcctgggga tttggggttg ctctggaaaa
1561 ctcatttgca ccactgctgt gccttggaat gctagttgga gtaataaatc tctggaacag
1621 atttggaatc acacgacctg gatggagtgg gacagagaaa ttaacaatta cacaagctta
1681 atacactcct taattgaaga atcgcaaaac cagcaagaaa agaatgaaca agaattattg
1741 gaattagata aatgggcaag tttgtggaat tggtttaaca taacaaattg gctgtggtat
1801 ataaaattat tcataatgat agtaggaggc ttggtaggtt taagaatagt ttttgctgta
1861 ctttctatag tgaatagagt taggcaggga tattcaccat tatcgtttca gacccacctc
1921 ccaaccccga ggggacccga caggcccgaa ggaatagaag aagaaggtgg agagagagac
1981 agagacagat ccattcgatt agtgaacgga tctcgacggt atcgccaaat ggcagtattc
2041 atccacaatt ttaaaagaaa aggggggatt ggggggtaca gtgcagggga aagaatagta
2101 gacataatag caacagacat acaaactaaa gaattacaaa aacaaattac aaaaattcaa
2161 aattttcggg tttattacag ggacagcaga gatccagttt ggatcgataa gcttgatatc
2221 gaattcctgc agccccgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
2281 tgaaagaccc cacctgtagg tttggcaagc tagctgcagt aacgccattt tgcaaggcat
2341 ggaaaaatac caaaccaaga atagagaagt tcagatcaag ggcgggtaca tgaaaatagc
2401 taacgttggg ccaaacagga tatctgcggt gagcagtttc ggccccggcc cggggccaag
2461 aacagatggt caccgcagtt tcggccccgg cccgaggcca agaacagatg gtccccagat
2521 atggcccaac cctcagcagt ttcttaagac ccatcagatg tttccaggct cccccaagga
2581 cctgaaatga ccctgcgcct tatttgaatt aaccaatcag cctgcttctc gcttctgttc
2641 gcgcgcttct gcttcccgag ctctataaaa gagctcacaa cccctcactc ggcgcgccag
2701 tcctccgaca gactgagtcg cccggggggg atctggagct ctcgagaatt ctcacgcgtc
2761 tgcaggatat caagcttgcg gtaccgcggg cccggccacc atggacaaga agtacagcat
2821 cggcctggcc atcggcacca actctgtggg ctgggccgtg atcaccgacg agtacaaggt
2881 gcccagcaag aaattcaagg tgctgggcaa caccgaccgg cacagcatca agaagaacct
2941 gatcggcgcc ctgctgttcg acagcggaga aacagccgag gccacccggc tgaagagaac
3001 cgccagaaga agatacacca gacggaagaa ccggatctgc tatctgcaag agatcttcag
3061 caacgagatg gccaaggtgg acgacagctt cttccacaga ctggaagagt ccttcctggt
3121 ggaagaggat aagaagcacg agcggcaccc catcttcggc aacatcgtgg acgaggtggc
3181 ctaccacgag aagtacccca ccatctacca cctgagaaag aaactggtgg acagcaccga
3241 caaggccgac ctgcggctga tctatctggc cctggcccac atgatcaagt tccggggcca
3301 cttcctgatc gagggcgacc tgaaccccga caacagcgac gtggacaagc tgttcatcca
3361 gctggtgcag acctacaacc agctgttcga ggaaaacccc atcaacgcca gcggcgtgga
3421 cgccaaggcc atcctgtctg ccagactgag caagagcaga cggctggaaa atctgatcgc
3481 ccagctgccc ggcgagaaga agaatggcct gttcggcaac ctgattgccc tgagcctggg
3541 cctgaccccc aacttcaaga gcaacttcga cctggccgag gatgccaaac tgcagctgag
3601 caaggacacc tacgacgacg acctggacaa cctgctggcc cagatcggcg accagtacgc
3661 cgacctgttt ctggccgcca agaacctgtc cgacgccatc ctgctgagcg acatcctgag
3721 agtgaacacc gagatcacca aggcccccct gagcgcctct atgatcaaga gatacgacga
3781 gcaccaccag gacctgaccc tgctgaaagc tctcgtgcgg cagcagctgc ctgagaagta
3841 caaagagatt ttcttcgacc agagcaagaa cggctacgcc ggctacatcg atggcggagc
3901 cagccaggaa gagttctaca agttcatcaa gcccatcctg gaaaagatgg acggcaccga
3961 ggaactgctc gtgaagctga acagagagga cctgctgcgg aagcagcgga ccttcgacaa
4021 cggcagcatc ccccaccaga tccacctggg agagctgcac gccattctgc ggcggcagga
4081 agatttttac ccattcctga aggacaaccg ggaaaagatc gagaagatcc tgaccttccg
4141 catcccctac tacgtgggcc ctctggccag gggaaacagc agattcgcct ggatgaccag
4201 aaagagcgag gaaaccatca ccccctggaa cttcgaggaa gtggtggaca agggcgccag
4261 cgcccagagc ttcatcgagc ggatgaccaa cttcgataag aacctgccca acgagaaggt
4321 gctgcccaag cacagcctgc tgtacgagta cttcaccgtg tacaacgagc tgaccaaagt
4381 gaaatacgtg accgagggaa tgagaaagcc cgccttcctg agcggcgagc agaaaaaagc
4441 catcgtggac ctgctgttca agaccaaccg gaaagtgacc gtgaagcagc tgaaagagga
4501 ctacttcaag aaaatcgagt gcttcgactc cgtggaaatc tccggcgtgg aagatcggtt
4561 caacgcctcc ctgggcacat accacgatct gctgaaaatt atcaaggaca aggacttcct
4621 ggacaatgag gaaaacgagg acattctgga agatatcgtg ctgaccctga cactgtttga
4681 ggacagagag atgatcgagg aacggctgaa aacctatgcc cacctgttcg acgacaaagt
4741 gatgaagcag ctgaagcggc ggagatacac cggctggggc aggctgagcc ggaagctgat
4801 caacggcatc cgggacaagc agtccggcaa gacaatcctg gatttcctga agtccgacgg
4861 cttcgccaac agaaacttca tgcagctgat ccacgacgac agcctgacct ttaaagagga
4921 catccagaaa gcccaggtgt ccggccaggg cgatagcctg cacgagcaca ttgccaatct
4981 ggccggcagc cccgccatta agaagggcat cctgcagaca gtgaaggtgg tggacgagct
5041 cgtgaaagtg atgggccggc acaagcccga gaacatcgtg atcgaaatgg ccagagagaa
5101 ccagaccacc cagaagggac agaagaacag ccgcgagaga atgaagcgga tcgaagaggg
5161 catcaaagag ctgggcagcc agatcctgaa agaacacccc gtggaaaaca cccagctgca
5221 gaacgagaag ctgtacctgt actacctgca gaatgggcgg gatatgtacg tggaccagga
5281 actggacatc aaccggctgt ccgactacga tgtggacgct atcgtgcctc agagctttct
5341 gaaggacgac tccatcgata acaaagtgct gactcggagc gacaagaacc ggggcaagag
5401 cgacaacgtg ccctccgaag aggtcgtgaa gaagatgaag aactactggc gccagctgct
5461 gaatgccaag ctgattaccc agaggaagtt cgacaatctg accaaggccg agagaggcgg
5521 cctgagcgaa ctggataagg ccggcttcat caagagacag ctggtggaaa cccggcagat
5581 cacaaagcac gtggcacaga tcctggactc ccggatgaac actaagtacg acgagaacga
5641 caaactgatc cgggaagtga aagtgatcac cctgaagtcc aagctggtgt ccgatttccg
5701 gaaggatttc cagttttaca aagtgcgcga gatcaacaac taccaccacg cccacgacgc
5761 ctacctgaac gccgtcgtgg gaaccgccct gatcaaaaag taccctaagc tggaaagcga
5821 gttcgtgtac ggcgactaca aggtgtacga cgtgcggaag atgatcgcca agagcgagca
5881 ggaaatcggc aaggctaccg ccaagtactt cttctacagc aacatcatga actttttcaa
5941 gaccgagatt accctggcca acggcgagat ccggaagcgg cctctgatcg agacaaacgg
6001 cgaaacaggc gagatcgtgt gggataaggg ccgggacttt gccaccgtgc ggaaagtgct
6061 gtctatgccc caagtgaata tcgtgaaaaa gaccgaggtg cagacaggcg gcttcagcaa
6121 agagtctatc ctgcccaaga ggaacagcga caagctgatc gccagaaaga aggactggga
6181 ccctaagaag tacggcggct tcgacagccc caccgtggcc tattctgtgc tggtggtggc
6241 caaagtggaa aagggcaagt ccaagaaact gaagagtgtg aaagagctgc tggggatcac
6301 catcatggaa agaagcagct tcgagaagaa tcccatcgac tttctggaag ccaagggcta
6361 caaagaagtg aaaaaggacc tgatcatcaa gctgcctaag tactccctgt tcgagctgga
6421 aaacggccgg aagagaatgc tggcctctgc cggcgaactg cagaagggaa acgaactggc
6481 cctgccctcc aaatatgtga acttcctgta cctggccagc cactatgaga agctgaaggg
6541 ctcccccgag gataatgagc agaaacagct gtttgtggaa cagcacaaac actacctgga
6601 cgagatcatc gagcagatca gcgagttctc caagagagtg atcctggccg acgctaatct
6661 ggacaaggtg ctgagcgcct acaacaagca cagagacaag cctatcagag agcaggccga
6721 gaatatcatc cacctgttta ccctgaccaa tctgggagcc cctgccgcct tcaagtactt
6781 tgacaccacc atcgaccgga agaggtacac cagcaccaaa gaggtgctgg acgccaccct
6841 gatccaccag agcatcaccg gcctgtacga gacacggatc gacctgtctc agctgggagg
6901 cgacgcctat ccctatgacg tgcccgatta tgccagcctg ggcagcggct cccccaagaa
6961 aaaacgcaag gtggaagatc ctaagaaaaa gcggaaagtg gacggcattg gtagtgggag
7021 caacggcagc agcggatcca gcgagctgat taaggagaac atgcacatga agctgtacat
7081 ggagggcacc gtggacaacc atcacttcaa gtgcacatcc gagggcgaag gcaagcccta
7141 cgagggcacc cagaccatga gaatcaaggt ggtcgagggc ggccctctcc ccttcgcctt
7201 cgacatcctg gctactagct tcctctacgg cagcaagacc ttcatcaacc acacccaggg
7261 catccccgac ttcttcaagc agtccttccc tgagggcttc acatgggaga gagtcaccac
7321 atacgaagac gggggcgtgc tgaccgctac ccaggacacc agcctccagg acggctgcct
7381 catctacaac gtcaagatca gaggggtgaa cttcacatcc aacggccctg tgatgcagaa
7441 gaaaacactc ggctgggagg ccttcaccga gacgctgtac cccgctgacg gcggcctgga
7501 aggcagaaac gacatggccc tgaagctcgt gggcgggagc catctgatcg caaacatcaa
7561 gaccacatat agatccaaga aacccgctaa gaacctcaag atgcctggcg tctactatgt
7621 ggactacaga ctggaaagaa tcaaggaggc caacaacgag acctacgtcg agcagcacga
7681 ggtggcagtg gccagatact gcgacctccc tagcaaactg gggcacaagc ttaattagta
7741 aggccgcgac tctagagtcg acctgcaggc atgcaagctt gatatcaagc ttatcgataa
7801 tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc
7861 ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat
7921 ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg
7981 gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg
8041 ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat
8101 tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt
8161 gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc
8221 ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa
8281 tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg
8341 ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcatc gataccgtcg
8401 acctcgaggg aattaattcg agctcggtac ctttaagacc aatgacttac aaggcagctg
8461 tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt cactcccaac
8521 gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag atctgagcct
8581 gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag
8641 tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac
8701 ccttttagtc agtgtggaaa atctctagca gcatctagaa ttaattccgt gtattctata
8761 gtgtcaccta aatcgtatgt gtatgataca taaggttatg tattaattgt agccgcgttc
8821 taacgacaat atgtacaagc ctaattgtgt agcatctggc ttactgaagc agaccctatc
8881 atctctctcg taaactgccg tcagagtcgg tttggttgga cgaaccttct gagtttctgg
8941 taacgccgtc ccgcacccgg aaatggtcag cgaaccaatc agcagggtca tcgctagcca
9001 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
9061 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
9121 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
9181 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
9241 ctgcttccta atgcaggagt cgcataaggg agagcgtcga atggtgcact ctcagtacaa
9301 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
9361 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
9421 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga aagggcctcg
9481 tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg
9541 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
9601 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
9661 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
9721 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
9781 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
9841 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
9901 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
9961 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
10021 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
10081 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
10141 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
10201 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
10261 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
10321 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
10381 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
10441 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
10501 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
10561 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
10621 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
10681 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
10741 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
10801 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgtctttcta gtgtagccgt
10861 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
10921 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
10981 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
11041 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
11101 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
11161 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
11221 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
11281 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
11341 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
11401 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
11461 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
11521 gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag
11581 tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc
11641 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct
11701 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
11761 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
11821 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagcttggac acaagacagg
11881 cttgcgagat atgtttgaga ataccacttt atcccgcgtc agggagaggc agtgcgtaaa
11941 aagacgcgga ctcatgtgaa atactggttt ttagtgcgcc agatctctat aatctcgcgc
12001 aacctatttt cccctcgaac actttttaag ccgtagataa acaggctggg acacttcaca
12061 tgagcgaaaa atacatcgtc acctgggaca tgttgcagat ccatgcacgt aaactcgcaa
12121 gccgactgat gccttctgaa caatggaaag gcattattgc cgtaagccgt ggcggtctgt
12181 accgggtgcg ttactggcgc gtgaactggg tattcgtcat gtcgataccg tttgtatttc
12241 cagctacgat cacgacaacc agcgcgagct taaagtgctg aaacgcgcag aaggcgatgg
12301 cgaaggcttc atcgttattg atgacctggt ggataccggt ggtactgcgg ttgcgattcg
12361 tgaaatgtat ccaaaagcgc actttgtcac catcttcgca aaaccggctg gtcgtccgct
12421 ggttgatgac tatgttgttg atatcccgca agatacctgg attgaacagc cgtgggatat
12481 gggcgtcgta ttcgtcccgc caatctccgg tcgctaatct tttcaacgcc tggcactgcc
12541 gggcgttgtt ctttttaact tcaggcgggt tacaatagtt tccagtaagt attctggagg
12601 ctgcatccat gacacaggca aacctgagcg aaaccctgtt caaaccccgc tttaaacatc
12661 ctgaaacctc gacgctagtc cgccgcttta atcacggcgc acaaccgcct gtgcagtcgg
12721 cccttgatgg taaaaccatc cctcactggt atcgcatgat taaccgtctg atgtggatct
12781 ggcgcggcat tgacccacgc gaaatcctcg acgtccaggc acgtattgtg atgagcgatg
12841 ccgaacgtac cgacgatgat ttatacgata cggtgattgg ctaccgtggc ggcaactgga
12901 tttatgagtg ggccccggat ctttgtgaag gaaccttact tctgtggtgt gacataattg
12961 gacaaactac ctacagagat ttaaagctct aaggtaaata taaaattttt aagtgtataa
13021 tgtgttaaac tactgattct aattgtttgt gtattttaga ttccaaccta tggaactgat
13081 gaatgggagc agtggtggaa tgcctttaat gaggaaaacc tgttttgctc agaagaaatg
13141 ccatctagtg atgatgaggc tactgctgac tctcaacatt ctactcctcc aaaaaagaag
13201 agaaaggtag aagaccccaa ggactttcct tcagaattgc taagtttttt gagtcatgct
13261 gtgtttagta atagaactct tgcttgcttt gctatttaca ccacaaagga aaaagctgca
13321 ctgctataca agaaaattat ggaaaaatat tctgtaacct ttataagtag gcataacagt
13381 tataatcata acatactgtt ttttcttact ccacacaggc atagagtgtc tgctattaat
13441 aactatgctc aaaaattgtg tacctttagc tttttaattt gtaaaggggt taataaggaa
13501 tatttgatgt atagtgcctt gactagagat cataatcagc cataccacat ttgtagaggt
13561 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
13621 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
13681 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
13741 catcaatgta tcttatcatg tctggatcaa ctggataact caagctaacc aaaatcatcc
13801 caaacttccc accccatacc ctattaccac tgccaattac ctagtggttt catttactct
13861 aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt taaatatgta
13921 ctacaaactt agtagt
//
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