质粒名称: | CLYBL-TO-hNGN2-BSD-mApple |
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出品公司: | Addgene |
目录编号: | 124229 |
存储实验室: | Michael Ward |
载体骨架基本信息 |
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载体名称: | CLYBL targeting |
载体出品公司: | Clontech |
载体类型: | 哺乳细胞表达 |
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 |
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插入基因名称: | NGN2、Neurogenin 2 |
物种来源: | H. sapiens (human) |
文献引用方法 |
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材料和方法部分: | CLYBL-TO-hNGN2-BSD-mApple was a gift from Michael Ward (Addgene plasmid # 124229 ; http://n2t.net/addgene:124229 ; RRID:Addgene_124229) |
参考文献部分: | CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons. Tian R, Gachechiladze MA, Ludwig CH, Laurie MT, Hong JY, Nathaniel D, Prabhu AV, Fernandopulle MS, Patel R, Abshari M, Ward ME, Kampmann M. Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 10.1016/j.neuron.2019.07.014. 10.1016/j.neuron.2019.07.014 PubMed 31422865 |
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CLYBL-TO-hNGN2-BSD-mApple载体质粒基本信息
CLYBL-TO-hNGN2-BSD-mApple质粒图谱载体图谱和CLYBL-TO-hNGN2-BSD-mApple载体序列质粒序列多克隆位点信息
CLYBL-TO-hNGN2-BSD-mApple质粒载体简介
CLYBL-TO-hNGN2-BSD-mApple质粒序列载体序列
如果没有序列信息,请删除本句话LOCUS CLYBL-TO-hNGN2-BSD-mApple 12234 bp DNA circular SYN 25-AUG-2024 DEFINITION Targets the human CLYBL safe harbor locus for integration of dox-inducible NGN2 for differentiation of iPSCs into cortical neurons. Contains a floxed BSD-NLS-mApple selection cassette.. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12234) AUTHORS Tian R, Gachechiladze MA, Ludwig CH, Laurie MT, Hong JY, Nathaniel D, Prabhu AV, Fernandopulle MS, Patel R, Abshari M, Ward ME, Kampmann M TITLE CRISPR Interference-Based Platform for Multimodal Genetic Screens in Human iPSC-Derived Neurons. JOURNAL Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 10.1016/j.neuron.2019.07.014. PUBMED 31422865 REFERENCE 2 (bases 1 to 12234) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 25, 2024 from SnapGene Server 7.0.3 https://www.biofeng.com COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Neuron. 2019 Aug 5. pii: S0896-6273(19)30640-3. doi: 10.1016/j.neuron.2019.07.014." FEATURES Location/Qualifiers source 1..12234 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 1..380 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 660..1676 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1600..1622 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1684..1703 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1735..2481 /codon_start=1 /product="modified rtTA protein that binds tightly to promoters containing the tet operator in the presence of doxycycline" /label=Tet-On(R) 3G /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA DALDDFDLDMLPADALDDFDLDMLPG" primer_bind complement(2572..2591) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 2637..2692 /label=beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(2691..2710) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" primer_bind 3002..3019 /label=L4440 /note="L4440 vector, forward primer" protein_bind 3136..3157 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 3172..3202 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 3210..3226 /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)." primer_bind 3215..3237 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 3234..3250 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 3234..3250 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" polyA_signal 3299..3420 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3310..3329) /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind 3364..3383 /label=SV40pA-R /note="SV40 polyA, reverse primer" promoter complement(4288..4665) /label=TRE3G promoter /note="3rd-generation Tet-responsive promoter that can be activated by binding of Tet-On(R) 3G" primer_bind complement(4355..4379) /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 4424..4442 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4460..4478 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4496..4514 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4532..4550 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4568..4586 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4604..4622 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" protein_bind 4640..4658 /gene="tetO" /label=tet operator /bound_moiety="tetracycline repressor TetR" /note="bacterial operator O2 for the tetR and tetA genes" rep_origin complement(6352..6940) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6432..6451) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7111..7971) /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 7734..7753 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" protein_bind complement(8829..8862) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." misc_feature 8863..8888 /label=SA /note="splice acceptor site" CDS 8912..8965 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 8960..9760 /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="GPRIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQG RPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDL LSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEE HQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQD IALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(9020..9039) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 9630..9649 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 9765..9989 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" promoter 10025..10236 /label=EF-1-alpha core promoter /note="core promoter for human elongation factor EF-1-alpha" LTR 10249..10517 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from human T-cell leukemia virus (HTLV) type 1" CDS 10986..11039 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 11046..11072 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" CDS 11088..11114 /codon_start=1 /product="nuclear localization signal of human c-Myc proto-oncogene (Dang and Lee, 1988)" /label=c-myc NLS /translation="PAAKRVKLD" regulatory 11163..11172 /label=Kozak sequence /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" /regulatory_class="other" protein_bind complement(11950..11983) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (ATGTATGC) (Shaw et al., 2021)." primer_bind complement(11999..12016) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 12005..12229 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" ORIGIN 1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc 61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca 121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga 181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc 241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct 301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat 361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct 421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga 481 tgggggcggg gggggggggg gggcgcgcgc cggggggggg gggggggggg gggggggggg 541 ggggggsgrg gsggrgrggk gcggcggcag ccaatcagag cggcgcgctc cgaaagtttc 601 cttttatggc gaggcggcgg cggcggcggc cctataaaaa gcgaagcgcg cggcgggcgg 661 gagtcgctgc gcgctgcctt cgccccgtgc cccgctccgc cgccgcctcg cgccgcccgc 721 cccggctctg actgaccgcg ttactcccac aggtgagcgg gcgggacggc ccttctcctc 781 cgggctgtaa ttagcgcttg gtttaatgac ggcttgtttc ttttctgtgg ctgcgtgaaa 841 gccttgaggg gctccgggag ggccctttgt gcggggggag cggctcgggg ggtgcgtgcg 901 tgtgtgtgtg cgtggggagc gccgcgtgcg gctccgcgct gcccggcggc tgtgagcgct 961 gcgggcgcgg cgcggggctt tgtgcgctcc gcagtgtgcg cgaggggagc gcggccgggg 1021 gcggtgcccc gcggtgcggg gggggctgcg aggggaacaa aggctgcgtg cggggtgtgt 1081 gcgtgggggg gtgagcaggg ggtgtgggcg cgtcggtcgg gctgcaaccc cccctgcacc 1141 cccctccccg agttgctgag cacggcccgg cttcgggtgc ggggctccgt acggggcgtg 1201 gcgcggggct cgccgtgccg ggcggggggt ggcggcaggt gggggtgccg ggcggggcgg 1261 ggccgcctcg ggccggggrg ggctcggggg rggggcgcgg cggcccccgg agcgccggcg 1321 gctgtcgagg cgcggcgagc cgcagccatt gccttttatg gtaatcgtgc gagagggcgc 1381 agggacttcc tttgtcccaa atctgtgcgg agccgaaatc tgggaggcgc cgccgcaccc 1441 cctctagcgg gcgcggggcg aagcggtgcg gcgccggcag gaaggaaatg ggcggggagg 1501 gccttcgtgc gtcgccgcgc cgccgtcccc ttctccctct ccagcctcgg ggctgtccgc 1561 ggggggacgg ctgccttcgg gggggacggg gcagggcggg gttcggcttc tggcgtgtga 1621 ccggcggctc tagagcctct gctaaccatg ttcatgcctt cttctttttc ctacagctcc 1681 tgggcaacgt gctggttatt gtgctgtctc atcattttgg caaagaatgt cgacatgagc 1741 cggctggaca agagcaaagt gatcaacagc gccctggaac tgctgaacgg cgtgggcatc 1801 gagggcctga ccacccggaa gctggcccag aaactgggcg tggaacagcc caccctgtac 1861 tggcacgtga agaacaagcg ggccctgctg gacgccctgc ccatcgagat gctggaccgg 1921 caccacaccc acagctgccc tctggaaggc gagagctggc aggacttcct gcggaacaac 1981 gccaagagct acagatgcgc cctgctgagc caccgggacg gcgccaaagt gcacctgggc 2041 accagaccca ccgagaagca gtacgagaca ctggaaaacc agctggcctt cctgtgccag 2101 cagggcttca gcctggaaaa cgccctgtac gccctgagcg ccgtgggcca cttcaccctg 2161 ggctgcgtgc tggaagaaca ggaacaccag gtcgccaaag aggaaagaga gacacccacc 2221 accgacagca tgccccccct gctgaagcag gccatcgagc tgttcgacag acagggcgcc 2281 gagcccgcct tcctgttcgg cctggaactg atcatctgcg gcctcgagaa gcagctgaag 2341 tgcgagagcg gcggacccac cgacgccctg gacgacttcg acctggacat gctgcccgcc 2401 gatgccctgg atgattttga tctggatatg ctccctgctg acgctctcga tgacttcgat 2461 ctcgatatgc tgccaggcta aaacgcgtga attcactcct caggtgcagg ctgcctatca 2521 gaaggtggtg actggtgtgg ccaatgccct ggctcacaaa taccactgag atctttttcc 2581 ctctgccaaa aattatgggg acatcatgaa gccccttgag catctgactt ctggctaata 2641 aaggaaattt attttcattg caatagtgtg ttggaatttt ttgtgtctct cactcggaag 2701 gacatatggg agggcaaatc atttaaaaca tcagaatgag tatttggttt agagtttggc 2761 aacatatgcc catatgctgg ctgccatgaa caaaggttgg ctataaagag gtcatcagta 2821 tatgaaacag ccccctgctg tccattcctt attccataga aaagccttga cttgaggtta 2881 gatttttttt atattttgtt ttgtgttatt tttttcttta acatccctaa aattttcctt 2941 acatgtgcgg ccgtcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg 3001 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc 3061 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag 3121 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt 3181 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa 3241 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata 3301 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt 3361 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt 3421 ggggtgggcg atcgattcgc gagtttaaac ttacaagata caatccctgg ctatggggag 3481 gtgaggtgca tagcggtgct tgtcgggagg tgggggctgc caatagtcca tgtctgaccc 3541 ggccgggctg gcgggcgata aagtgcagct gtagggggag gtggaattgg aggacacgga 3601 ggaggacggc gcggggctgt tggtgcaact ccacgtggag gcgggcgagg ggctgtctcc 3661 gctgctgctc agggcggcgc tggcgcctcc cgggctcagc aacactgcct cggagaagag 3721 cgcccccggc aggcccccgc cgccgccccc gcagtgatcc gccaggcgca gggtctcggt 3781 gagtgcccag atgtagttgt gggcgaagcg cagggtctcg atcttggtga gcttggcgtc 3841 ctcggggaac gtggggagca cctcgcgcag cgcgtccagt gccgcgttga ggttgtgcat 3901 gcggtttcgc tcgcggttgt tggccttcag tctacgggtc ttcttgatgc gctgcaccgt 3961 ctcggccgtc ttggcgcctc gggagacggc ccgcgcccgg gaagggcgcc gtttgcaatc 4021 gtgtaccaga cccagcagcc gtgcgggccg gcagccctcc gcacccgcag ccacgccgcc 4081 ccgcgccccc tgcccggcct cagccccgcg ctgccgacgc gccccgcctg acgcgcccgg 4141 ctcctcctcc tcttcttcgt cggcgctgga tgacagcggg gtcagggccg ccaaggcggg 4201 ggaggccgat ccgagcagca ctaacacgtc ctcttcctcc ttcaactcca aggtctcgga 4261 tttgacgaac atggtggcgg cggatccttt acgagggtag gaagtggtac ggaaagttgg 4321 tataagacaa aagtgttgtg gaattgctcc aggcgatctg acggttcact aaacgagctc 4381 tgcttttata ggcgcccacc gtacacgcct aaagcttata cgttctctat cactgatagg 4441 gagtaaactg gatatacgtt ctctatcact gatagggagt aaactgtaga tacgttctct 4501 atcactgata gggagtaaac tggtcatacg ttctctatca ctgataggga gtaaactcct 4561 tatacgttct ctatcactga tagggagtaa agtctgcata cgttctctat cactgatagg 4621 gagtaaactc ttcatacgtt ctctatcact gatagggagt aaactggtac cctccgcgcc 4681 tacagctcaa gccacatccg aagggggagg gagccgggag ctgcgcgcgg ggccgccggg 4741 gggaggggtg gcaccgccca cgccgggcgg ccacgaaggg cggggcagcg ggcgcgcgcc 4801 cggcgggggg aggggccgcg cgccgcgccc gctgggaatt ggggccctag ggggagggcg 4861 gaggcgccga cgaccgcggc acttaccgtt cgcggcgtgg cgcccggtgg tccccaaggg 4921 gagggaaggg ggaggcgggg cgaggacagt gaccggagtc tcctcagcgg tggcttttct 4981 gcttggcagc ctcagcggct ggcgccaaaa ccggactccg cccacttcct cgcccctgcg 5041 gtgcgagggt gtggaatcct ccagacgctg ggggaggggg agttgggagc ttaaaaacta 5101 gtaccccttt gggaccactt tcagcagcga actctcctgt acaccagggg tcagttccac 5161 agacgcgggc caggggtggg tcattgcggc gtgaacaata atttgactag aagttgattc 5221 gggtgtttcc ggaaggggcc gagtcaatcc gccgagttgg ggcacggaaa acaaaaaggg 5281 aaggctacta agatttttct ggcgggggtt atcattggcg taactgcagg gaccacctcc 5341 cttaattaaa tctcctcatc cttccaacat aaatatattt tgggattata tcaacattca 5401 atgttactta aagtgacctt gtaaatattt tcacaactga gccatgtttg atttgtatac 5461 ttatgtttac tttactgttt ttcctgaagt taataattgc cttgaattta tttatttctt 5521 taaaaatgtt tcattactca ggactgtagt ttacattacg attctttgtg ttatacagtt 5581 gatgggtttc ttttctttct taatttcttt aaaaaataga gatggggtct tactatatta 5641 cccaggctgg tcttgaagtc ctgggctcaa gtgatcttcc tgtctcagcc taccaagtag 5701 ctgagactat aggtgcaaaa aagccactat acctggctag tttacaggtt ttaacaaatg 5761 cattatgcca cgtatccatt attacaggat cacacaagat attttcatta ccctgaaagc 5821 atccctgtgt tccaccaatt catcctgcct ccatgagccg ctggcaacca ctgatctcta 5881 tagttttgcc ttttctaaaa tgtcatataa ttggaatcat acagtctgta gcattttcag 5941 actagctttt aaaatttggc aatatgcatt taaggttcct ccttaaatgt gaagggcata 6001 gccaatgtgg cttgatagct catttctttt tattggtgaa tatttcattg tctggatgtt 6061 ccacagtttg tttatccatt cacctattca atttgctttt tttctgtgta tctatcacta 6121 attcaatact ggactctcca acagagccgt cccgcttccc taaggccctg ctctgggctt 6181 ctgggtttga gtccttggca agcccaggag aggcgctcag gcttccctgt cccccttcct 6241 cgtccaccat ctcatgcccc tggctctcct gccccttccc tacaggggtt cctggctctg 6301 ctctaagggc gaattcgcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag 6361 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc 6421 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt 6481 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct 6541 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg 6601 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct 6661 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat 6721 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg 6781 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa 6841 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt 6901 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc 6961 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt 7021 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta 7081 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat 7141 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac 7201 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg 7261 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag 7321 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt 7381 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt 7441 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt 7501 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt 7561 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct 7621 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt 7681 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac 7741 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa 7801 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa 7861 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca 7921 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct 7981 ttttcaatag caggagaatc atgccaatgg gccaatatac attctgaccc acagtttcat 8041 aataaaataa aatggttgtg gttgtaagcc actatgtttc agagtggttt gttacacagc 8101 aataaataac taatatagta ggcataccat caagtccaaa gtaggtagag aagaatgtaa 8161 atagcagagc aaaacagcat gactggtggc tgggaggctt aaaactggga caggatcaga 8221 gtcatgaaag aagtcaaaga aatggttcag aagtaaggct gagactgact tacaaaagct 8281 gaaagtccct ttaagttggt gtttggtgca ttggcagggg caggtatggt gacttaaaag 8341 agccatgctc aacaagatca agcacaacac aatcacgggt caccccagca gaccttagcg 8401 agtctagcca tttctttggt ggtggtcaca gtcatgcttc agcccagttt ccacttggac 8461 aaatggtaca tattttcaat gagatgaaaa ttaagataca atccatgtgc tcagagagtg 8521 atcacagctc tgactaaaca ctgtgcccca aagtgttgag gaattgggaa aacctagctg 8581 agttagtggt ctcttttctg ttacaataaa gctcataatg aaaattagcc ttctttgttc 8641 ttccccaagt ctttctttct agacgaaact actttcaact gttttaactt ccttactgtt 8701 aacttccata tttctagata gtatggtcag actgttattt cttgactttt aaatgtaaga 8761 tattatctac tgacttcctt ctatgtaaga tgaggattga gctctcttac ccttctccca 8821 gcggccgcat aacttcgtat agcatacatt atacgaagtt atctgacctc ttctcttcct 8881 cccacagggc ctcgagagat ctggcagcgg agagggcaga ggaagtcttc taacatgcgg 8941 tgacgtggag gagaatcccg gccctaggat tgaacaagat ggattgcacg caggttctcc 9001 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 9061 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 9121 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 9181 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 9241 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 9301 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 9361 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 9421 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 9481 caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg 9541 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 9601 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 9661 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 9721 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctaa aaacctgtgc cttctagttg 9781 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc 9841 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc 9901 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag 9961 gcatgctggg gatgcggtgg gctctatgga tgcatgatct gcgatcgctc cggtgcccgt 10021 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat 10081 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg 10141 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac 10201 gttctttttc gcaacgggtt tgccgccaga acacagctga agcttcgagg ggctcgcatc 10261 tctccttcac gcgcccgccg ccctacctga ggccgccatc cacgccggtt gagtcgcgtt 10321 ctgccgcctc ccgcctgtgg tgcctcctga actgcgtccg ccgtctaggt aagtttaaag 10381 ctcaggtcga gaccgggcct ttgtccggcg ctcccttgga gcctacctag actcagccgg 10441 ctctccacgc tttgcctgac cctgcttgct caactctacg tctttgtttc gttttctgtt 10501 ctgcgccgtt acagatccaa gctgtgaccg gcgcctacgc tagcgccacc atgaagacct 10561 tcaacatctc ccagcaggat ctagaattag tagaagtagc gacagagaag attacaatgc 10621 tttatgagga taataaacat catgtgggag cggcaattcg tacgaaaaca ggagaaatca 10681 tttcggcagt acatattgaa gcgtatatag gacgagtaac tgtttgtgca gaagccattg 10741 cgattggtag tgcagtttcg aatggacaaa aggattttga cacgattgta gctgttagac 10801 acccttattc tgacgaagta gatagaagta ttcgagtggt aagtccttgt ggtatgtgta 10861 gggagttgat ttcagactat gcaccagatt gttttgtgtt aatagaaatg aatggcaagt 10921 tagtcaaaac tacgattgaa gaactcattc cactcaaata tacccgaaat ggatcgggat 10981 cgggagaggg cagaggaagt ctgctaacat gcggtgacgt cgaggagaat cctggcccac 11041 cggtccctgc cgctaaacgg gtcaaactcg acggcggttc tggaggtcct gctgcaaagc 11101 gcgttaagtt ggatggaggg tcagggggag ccaccgccac cccggtcgcc acgctaccgg 11161 tcgccaccat ggtgtccaag ggcgaagaga acaacatggc catcatcaaa gaattcatgc 11221 ggttcaaggt gcacatggaa ggcagcgtga acggccacga gttcgagatt gaaggcgaag 11281 gcgagggcag accctacgag gcttttcaga ccgccaagct gaaagtgacc aaaggcggcc 11341 ctctgccttt tgcctgggac attctgagcc ctcagtttat gtacggcagc aaggtgtaca 11401 tcaagcaccc cgccgacatt cccgactact tcaagctgag cttccccgag ggcttcagat 11461 gggagagagt gatgaacttc gaggacggcg gcatcatcca cgtgaaccag gatagctctc 11521 tgcaggacgg cgtgttcatc tacaaagtga agctgcgggg caccaacttt ccctctgatg 11581 gccccgtgat gcagaaaaag accatgggct gggaagccag cgaggaaaga atgtaccctg 11641 aggatggcgc cctgaagtcc gagatcaaga agcggctgaa gctgaaggat ggcggccact 11701 atgccgccga agtgaaaacc acctacaagg ccaagaaacc cgtgcagctg cctggcgcct 11761 acatcgtgga tatcaagctg gacatcgtgt cccacaacga ggactacacc atcgtggaac 11821 agtacgagag agccgaaggc agacacagca caggcggaat ggacgagctg tacaagtaac 11881 accatcgtgg aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag 11941 taaggtacca taacttcgta tagcatacat tatacgaagt tataaacccg ctgatcagcc 12001 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg 12061 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat 12121 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag 12181 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggg tctc //
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