质粒名称: | pR26 CAG/GFP Dest |
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出品公司: | addgene |
目录编号: | 74281 |
存储实验室: | Ralf Kuehn |
载体骨架基本信息 |
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载体名称: | Bluescript |
完整质粒大小: | 16896 bp |
载体类型: | Mouse Targeting |
载体抗性: | Chloramphenicol and Ampicillin |
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: | Rosa26 5-homology region |
物种来源: | M. musculus (mouse) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | I-SceI(未破坏) |
3’克隆位点: | XbaI (未破坏) |
5’测序引物 | M13 forward |
3’测序引物 | M13 reverse |
文献引用方法 |
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材料和方法部分: | pR26 CAG/GFP Dest was a gift from Ralf Kuehn (Addgene plasmid # 74281 ; http://n2t.net/addgene:74281 ; RRID:Addgene_74281) |
参考文献部分: | Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes. Chu VT, Weber T, Graf R, Sommermann T, Petsch K, Sack U, Volchkov P, Rajewsky K, Kuhn R. BMC Biotechnol. 2016 Jan 16;16(1):4. doi: 10.1186/s12896-016-0234-4. 10.1186/s12896-016-0234-4 PubMed 26772810 |
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 pR26 CAG/GFP Dest 16896 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Gene targeting vector for the mouse Rosa26 locus, including a CAG promoter, loxP flanked stop cassette, GFP, for gateway cloning. ACCESSION . VERSION . KEYWORDS pR26 CAG/GFP Dest SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16896) AUTHORS Chu VT, Weber T, Graf R, Sommermann T, Petsch K, Sack U, Volchkov P, Rajewsky K, Kuhn R TITLE Efficient generation of Rosa26 knock-in mice using CRISPR/Cas9 in C57BL/6 zygotes. JOURNAL BMC Biotechnol. 2016 Jan 16;16(1):4. doi: 10.1186/s12896-016-0234-4. PUBMED 26772810 REFERENCE 2 (bases 1 to 16896) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.biofeng.com FEATURES Location/Qualifiers source 1..16896 /organism="synthetic DNA construct" /mol_type="other DNA" misc_feature 6..1088 /label=Rosa26 left arm /note="left homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)" misc_feature 1095..1198 /label=Ad SA /note="adenovirus major late transcript splice acceptor (Friedrich and Soriano, 1991)" polyA_signal 1217..1424 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" polyA_signal 1438..1645 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" enhancer 1654..2033 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" intron 2309..3325 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 3249..3271 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 3333..3352 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 3388..3421 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 3428..3476 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" CDS complement(3483..4286) /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="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3594..3613) /label=Neo-F /note="Neomycin resistance gene, forward primer" primer_bind 4204..4223 /label=Neo-R /note="Neomycin resistance gene, reverse primer" intron 5099..5164 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" polyA_signal 5739..5873 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5776..5795) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5830..5849 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind 5893..5926 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 5939..6063 /gene="mutant version of attR" /label=attR1 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" promoter 6088..6118 /label=lac UV5 promoter /note="E. coli lac promoter with an ""up"" mutation" CDS 6172..6831 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(6238..6257) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" CDS 7173..7478 /codon_start=1 /gene="ccdB" /product="CcdB, a bacterial toxin that poisons DNA gyrase" /label=ccdB /note="Plasmids containing the ccdB gene cannot be propagated in standard E. coli strains." /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 7357..7376 /label=ccdB-fwd /note="ccdB gene, forward primer" protein_bind complement(7519..7643) /gene="mutant version of attR" /label=attR2 /bound_moiety="LR Clonase(TM)" /note="recombination site for the Gateway(R) LR reaction" protein_bind 7678..7725 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." misc_feature 7764..8316 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(7908..7925) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 8135..8154 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 8328..9047 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 8328..9044 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(8373..8394) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(8634..8653) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 8981..9002 /label=EGFP-C /note="EGFP, forward primer" protein_bind 9072..9119 /label=FRT /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="FLP-mediated recombination occurs in the 8-bp core sequence TCTAGAAA (Turan and Bode, 2011)." primer_bind complement(9144..9160) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9144..9160) /label=pBluescriptSK /note="For pBluescript vector" primer_bind complement(9173..9190) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 9179..9403 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" misc_feature 9493..13833 /label=Rosa26 right arm /note="right homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)" primer_bind complement(13918..13941) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind complement(14051..14071) /label=T3 /note="T3 promoter, forward primer" promoter complement(14051..14069) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(14090..14106) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(14090..14106) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(14103..14125) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 14114..14130 /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(14138..14168) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 14183..14204 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(14321..14338) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(14492..15080) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(14572..14591) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(15251..16111) /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 15874..15893 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(16112..16216) /gene="bla" /label=AmpR promoter rep_origin complement(16269..16697) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(16379..16400) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 16591..16610 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 16824..16846 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 16838..16855 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 16839..16855 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 16865..16884 /label=T7 /note="T7 promoter, forward primer" promoter 16865..16883 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ccaccgcggc aggccctccg agcgtggtgg agccgttctg tgagacagcc gggtacgagt 61 cgtgacgctg gaaggggcaa gcgggtggtg ggcaggaatg cggtccgccc tgcagcaacc 121 ggagggggag ggagaaggga gcggaaaagt ctccaccgga cgcggccatg gctcgggggg 181 gggggggcag cggaggagcg cttccggccg acgtctcgtc gctgattggc ttcttttcct 241 cccgccgtgt gtgaaaacac aaatggcgtg ttttggttgg cgtaaggcgc ctgtcagtta 301 acggcagccg gagtgcgcag ccgccggcag cctcgctctg cccactgggt ggggcgggag 361 gtaggtgggg tgaggcgagc tggacgtgcg ggcgcggtcg gcctctggcg gggcgggggg 421 ggggagggag ggtcagcgaa agtagctcgc gcgcgagcgg ccgcccaccc tccccttcct 481 ctgggggagt cgttttaccc gccgccggcc gggcctcgtc gtctgattgg ctctcggggc 541 ccagaaaact ggcccttgcc attggctcgt gttcgtgcaa gttgagtcca tccgccggcc 601 agcgggggcg gcgaggaggc gctcccaggt tccggccctc ccctcggccc cgcgccgcag 661 agtctggccg cgcgcccctg cgcaacgtgg caggaagcgc gcgctggggg cggggacggg 721 cagtagggct gagcggctgc ggggcgggtg caagcacgtt tccgacttga gttgcctcaa 781 gaggggcgtg ctgagccaga cctccatcgc gcactccggg gagtggaggg aaggagcgag 841 ggctcagttg ggctgttttg gaggcaggaa gcacttgctc tcccaaagtc gctctgagtt 901 gttatcagta agggagctgc agtggagtag gcggggagaa ggccgcaccc ttctccggag 961 gggggagggg agtgttgcaa tacctttctg ggagttctct gctgcctcct ggcttctgag 1021 gaccgccctg ggcctgggag aatcccttcc ccctcttccc tcgtgatctg caactccagt 1081 ctttctagca tctgtagggc gcagtagtcc agggtttcct tgatgatgtc atacttatcc 1141 tgtccctttt ttttccacag ctcgcggttg aggacaaact cttcgcggtc tttccagtgg 1201 ttaattaact cgaggactgt gccttctagt tgccagccat ctgttgtttg cccctccccc 1261 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa 1321 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac 1381 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt ctagcgactg 1441 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg 1501 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga 1561 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg 1621 aagacaatag caggcatgct ggggatgcgg gtcgacattg attattgact agttattaat 1681 agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac 1741 ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa 1801 tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt 1861 atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc 1921 ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat 1981 gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg 2041 tgagccccac gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt 2101 atttatttat tttttaatta ttttgtgcag cgatgggggc gggggggggg ggcgcgcgcc 2161 aggcgggggg gggggggggg gggggggggg gggggggggg gggggggggg cggcggcagc 2221 caatcagagc ggcgcgctcc gaaagtttcc ttttatggcg aggcggcggc ggcggcggcc 2281 ctataaaaag cgaagcgcgc ggcgggcggg agtcgctgcg cgctgccttc gccccgtgcc 2341 ccgctccgcc gccgcctcgc gccgcccgcc ccggctctga ctgaccgcgt tactcccaca 2401 ggtgagcggg cgggacggcc cttctcctcc gggctgtaat tagcgcttgg tttaatgacg 2461 gcttgtttct tttctgtggc tgcgtgaaag ccttgagggg ctccgggagg gccctttgtg 2521 cggggggagc ggctcggggg gtgcgtgcgt gtgtgtgtgc gtggggagcg ccgcgtgcgg 2581 ctccgcgctg cccggcggct gtgagcgctg cgggcgcggc gcggggcttt gtgcgctccg 2641 cagtgtgcgc gaggggagcg cggccggggg cggtgccccg cggtgcgggg ggggctgcga 2701 ggggaacaaa ggctgcgtgc ggggtgtgtg cgtggggggg tgagcagggg gtgtgggcgc 2761 gtcggtcggg ctgcaacccc ccctgcaccc ccctccccga gttgctgagc acggcccggc 2821 ttcgggtgcg gggctccgta cggggcgtgg cgcggggctc gccgtgccgg gcggggggtg 2881 gcggcaggtg ggggtgccgg gcggggcggg gccgcctcgg gccggggagg gctcggggga 2941 ggggcgcggc ggcccccgga gcgccggcgg ctgtcgaggc gcggcgagcc gcagccattg 3001 ccttttatgg taatcgtgcg agagggcgca gggacttcct ttgtcccaaa tctgtgcgga 3061 gccgaaatct gggaggcgcc gccgcacccc ctctagcggg cgcggggcga agcggtgcgg 3121 cgccggcagg aaggaaatgg gcggggaggg ccttcgtgcg tcgccgcgcc gccgtcccct 3181 tctccctctc cagcctcggg gctgtccgcg gggggacggc tgccttcggg ggggacgggg 3241 cagggcgggg ttcggcttct ggcgtgtgac cggcggctct agagcctctg ctaaccatgt 3301 tcatgccttc ttctttttcc tacagctcct gggcaacgtg ctggttattg tgctgtctca 3361 tcattttggc aaagaattct taattaaata acttcgtata gcatacatta tacgaagtta 3421 tggatccgaa caaacgaccc aacacccgtg cgttttattc tgtcttttta ttgccgatcc 3481 cctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg 3541 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca 3601 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg 3661 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc 3721 acgacgagat cctcgccgtc gggcatgcgc gccttgagcc tggcgaacag ttcggctggc 3781 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga 3841 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca 3901 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg 3961 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct 4021 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc 4081 cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga 4141 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt 4201 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat 4261 ccatcttgtt caatggccga tcccatattg gctgcagggt cgctcggtgt tcgaggccac 4321 acgcgtcacc ttaatatgcg aagtggacct gggaccgcgc cgccccgact gcatctgcgt 4381 gttcgaattc gccaatgaca agacgctggg cggggtttgc tcgacattgg gtggaaacat 4441 tccaggcctg ggtggagagg ctttttgctt cctcttgcaa aaccacactg ctcgacattg 4501 ggtggaaaca ttccaggcct gggtggagag gctttttgct tcctcttgaa aaccacactg 4561 ctcgactagt gattaaagtc gactcgggga caccaaatat ggcgatctcg gccttttcgt 4621 ttcttggagc tgggacatgt ttgccatcga tccatctacc accagaacgg ccgttagatc 4681 tgctgccacc gttgtttcca ccgaagaaac caccgttgcc gtaaccacca cgacggttgt 4741 tgctaaagaa gctgccaccg ccacggccac cgttgtagcc gccgttgttg ttattgtagt 4801 tgctactgtt atttctggca cttcttggtt ttcctcttaa gtgaggagga acataaccat 4861 tctcgttgtt gtcgttgatg cttaaatttt gcacttgttc gctcagttca gccataatat 4921 gaaatgcttt tcttgttgtt cttacggaat accacttgcc acctatcacc acaactaact 4981 ttttcccgtt cctccatctc ttttatattt tttttctcga gggatctttg tgaaggaacc 5041 ttacttctgt ggtgtgacat aattggacaa actacctaca gagatttaaa gctctaaggt 5101 aaatataaaa tttttaagtg tataatgtgt taaactactg attctaattg tttgtgtatt 5161 ttagattcca acctatggaa ctgatgaatg ggagcagtgg tggaatgcct ttaatgagga 5221 aaacctgttt tgctcagaag aaatgccatc tagtgatgat gaggctactg ctgactctca 5281 acattctact cctccaaaaa agaagagaag ggtagaagac cccaaggact ttccttcaga 5341 attgctaagt tttttgagtc atgctgtgtt tagtaataga actcttgctt gctttgctat 5401 ttacaccaca aaggaaaaag ctgcactgct atacaagaaa attatggaaa aatattctgt 5461 aacctttata agtaggcata acagttataa tcataacata ctgttttttc ttactccaca 5521 caggcataga gtgtctgcta ttaataacta tgctcaaaaa ttgtgtacct ttagcttttt 5581 aatttgtaaa ggggttaata aggaatattt gatgtatagt gccttgacta gagatcataa 5641 tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc 5701 tgaacctgaa acatagaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata 5761 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc 5821 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atctgacatg 5881 gtaagtaagc ttataacttc gtatagcata cattatacga agttatcggc gcgccatcac 5941 aagtttgtac aaaaaagctg aacgagaaac gtaaaatgat ataaatatca atatattaaa 6001 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tccagtcact 6061 atggcggccg cattaggcac cccaggcttt acactttatg cttccggctc gtataatgtg 6121 tggattttga gttaggatcc gtcgagattt tcaggagcta aggaagctaa aatggagaaa 6181 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag 6241 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc 6301 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt 6361 gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg tgagctggtg 6421 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca 6481 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat 6541 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt 6601 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg 6661 gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg cgacaaggtg 6721 ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca tgtcggcaga 6781 atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta aacgcgtgga 6841 tccggcttac taaaagccag ataacagtat gcgtatttgc gcgctgattt ttgcggtata 6901 agaatatata ctgatatgta tacccgaagt atgtcaaaaa gaggtatgct atgaagcagc 6961 gtattacagt gacagttgac agcgacagct atcagttgct caaggcatat atgatgtcaa 7021 tatctccggt ctggtaagca caaccatgca gaatgaagcc cgtcgtctgc gtgccgaacg 7081 ctggaaagcg gaaaatcagg aagggatggc tgaggtcgcc cggtttattg aaatgaacgg 7141 ctcttttgct gacgagaaca ggggctggtg aaatgcagtt taaggtttac acctataaaa 7201 gagagagccg ttatcgtctg tttgtggatg tacagagtga tattattgac acgcccgggc 7261 gacggatggt gatccccctg gccagtgcac gtctgctgtc agataaagtc tcccgtgaac 7321 tttacccggt ggtgcatatc ggggatgaaa gctggcgcat gatgaccacc gatatggcca 7381 gtgtgccggt ctccgttatc ggggaagaag tggctgatct cagccaccgc gaaaatgaca 7441 tcaaaaacgc cattaacctg atgttctggg gaatataaat gtcaggctcc cttatacaca 7501 gccagtctgc aggtcgacca tagtgactgg atatgttgtg ttttacagta ttatgtagtc 7561 tgttttttat gcaaaatcta atttaatata ttgatattta tatcatttta cgtttctcgt 7621 tcagctttct tgtacaaagt ggtgatggcg cgccgcggcc gcgatcaatt cggtaccgaa 7681 gttcctattc cgaagttcct attctctaga aagtatagga acttcctcga gggtacccca 7741 attccgcccc cccccccccc ctaacgttac tggccgaagc cgcttggaat aaggccggtg 7801 tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg tgagggcccg 7861 gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc tcgccaaagg 7921 aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt cttgaagaca 7981 aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg acaggtgcct 8041 ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac cccagtgcca 8101 cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg tattcaacaa 8161 ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg ggcctcggtg 8221 cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc cgaaccacgg 8281 ggacgtggtt ttcctttgaa aaacacgatg ataatatggc cacaaccatg gtgagcaagg 8341 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg 8401 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc 8461 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc 8521 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct 8581 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg 8641 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg 8701 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca 8761 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga 8821 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc 8881 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc 8941 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg 9001 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaatcc gggatccgga 9061 gagctcccaa cgaagttcct attccgaagt tcctattctc tagaaagtat aggaacttcc 9121 tcgaggttgg atgcagcccg ggggatccac tagttctaga gcggccgatc agcctcgact 9181 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg 9241 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg 9301 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg 9361 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga 9421 accagctggg gctcgagatc cactagttct agcctcgagg ctagagcggc cgccgcgccg 9481 atatcgaatt cgctagaaga tgggcgggag tcttctgggc aggcttaaag gctaacctgg 9541 tgtgtgggcg ttgtcctgca ggggaattga acaggtgtaa aattggaggg acaagacttc 9601 ccacagattt tcggttttgt cgggaagttt tttaataggg gcaaataagg aaaatgggag 9661 gataggtagt catctggggt tttatgcagc aaaactacag gttattattg cttgtgatcc 9721 gcctcggagt attttccatc gaggtagatt aaagacatgc tcacccgagt tttatactct 9781 cctgcttgag atccttacta cagtatgaaa ttacagtgtc gcgagttaga ctatgtaagc 9841 agaattttaa tcatttttaa agagcccagt acttcatatc catttctccc gctccttctg 9901 cagccttatc aaaaggtatt ttagaacact cattttagcc ccattttcat ttattatact 9961 ggcttatcca acccctagac agagcattgg cattttccct ttcctgatct tagaagtctg 10021 atgactcatg aaaccagaca gattagttac atacaccaca aatcgaggct gtagctgggg 10081 cctcaacact gcagttcttt tataactcct tagtacactt tttgttgatc ctttgccttg 10141 atccttaatt ttcagtgtct atcacctctc ccgtcaggtg gtgttccaca tttgggccta 10201 ttctcagtcc agggagtttt acaacaatag atgtattgag aatccaacct aaagcttaac 10261 tttccactcc catgaatgcc tctctccttt ttctccattt ataaactgag ctattaacca 10321 ttaatggttt ccaggtggat gtctcctccc ccaatattac ctgatgtatc ttacatattg 10381 ccaggctgat attttaagac attaaaaggt atatttcatt attgagccac atggtattga 10441 ttactgctta ctaaaatttt gtcattgtac acatctgtaa aaggtggttc cttttggaat 10501 gcaaagttca ggtgtttgtt gtctttcctg acctaaggtc ttgtgagctt gtattttttc 10561 tatttaagca gtgctttctc ttggactggc ttgactcatg gcattctaca cgttattgct 10621 ggtctaaatg tgattttgcc aagcttcttc aggacctata attttgcttg acttgtagcc 10681 aaacacaagt aaaatgatta agcaacaaat gtatttgtga agcttggttt ttaggttgtt 10741 gtgttgtgtg tgcttgtgct ctataataat actatccagg ggctggagag gtggctcgga 10801 gttcaagagc acagactgct cttccagaag tcctgagttc aattcccagc aaccacatgg 10861 tggctcacaa ccatctgtaa tgggatctga tgccctcttc tggtgtgtct gaagaccaca 10921 agtgtattca cattaaataa ataaatcctc cttcttcttc tttttttttt ttttaaagag 10981 aatactgtct ccagtagaat ttactgaagt aatgaaatac tttgtgtttg ttccaatatg 11041 gtagccaata atcaaattac tctttaagca ctggaaatgt taccaaggaa ctaattttta 11101 tttgaagtgt aactgtggac agaggagcca taactgcaga cttgtgggat acagaagacc 11161 aatgcagact ttaatgtctt ttctcttaca ctaagcaata aagaaataaa aattgaactt 11221 ctagtatcct atttgtttaa actgctagct ttacttaact tttgtgcttc atctatacaa 11281 agctgaaagc taagtctgca gccattacta aacatgaaag caagtaatga taattttgga 11341 tttcaaaaat gtagggccag agtttagcca gccagtggtg gtgcttgcct ttatgccttt 11401 aatcccagca ctctggaggc agagacaggc agatctctga gtttgagccc agcctggtct 11461 acacatcaag ttctatctag gatagccagg aatacacaca gaaaccctgt tggggagggg 11521 ggctctgaga tttcataaaa ttataattga agcattccct aatgagccac tatggatgtg 11581 gctaaatccg tctacctttc tgatgagatt tgggtattat tttttctgtc tctgctgttg 11641 gttgggtctt ttgacactgt gggctttctt taaagcctcc ttcctgccat gtggtctctt 11701 gtttgctact aacttcccat ggcttaaatg gcatggcttt ttgccttcta agggcagctg 11761 ctgagatttg cagcctgatt tccagggtgg ggttgggaaa tctttcaaac actaaaattg 11821 tcctttaatt ttttttttaa aaaatgggtt atataataaa cctcataaaa tagttatgag 11881 gagtgaggtg gactaatatt aaatgagtcc ctcccctata aaagagctat taaggctttt 11941 tgtcttatac ttaacttttt ttttaaatgt ggtatcttta gaaccaaggg tcttagagtt 12001 ttagtataca gaaactgttg catcgcttaa tcagattttc tagtttcaaa tccagagaat 12061 ccaaattctt cacagccaaa gtcaaattaa gaatttctga cttttaatgt taatttgctt 12121 actgtgaata taaaaatgat agcttttcct gaggcagggt ctcactatgt atctctgcct 12181 gatctgcaac aagatatgta gactaaagtt ctgcctgctt ttgtctcctg aatactaagg 12241 ttaaaatgta gtaatacttt tggaacttgc aggtcagatt cttttatagg ggacacacta 12301 agggagcttg ggtgatagtt ggtaaaatgt gtttcaagtg atgaaaactt gaattattat 12361 caccgcaacc tactttttaa aaaaaaaagc caggcctgtt agagcatgct taagggatcc 12421 ctaggacttg ctgagcacac aagagtagtt acttggcagg ctcctggtga gagcatattt 12481 caaaaaacaa ggcagacaac caagaaacta cagttaaggt tacctgtctt taaaccatct 12541 gcatatacac agggatatta aaatattcca aataatattt cattcaagtt ttcccccatc 12601 aaattgggac atggatttct ccggtgaata ggcagagttg gaaactaaac aaatgttggt 12661 tttgtgattt gtgaaattgt tttcaagtga tagttaaagc ccatgagata cagaacaaag 12721 ctgctatttc gaggtctctt ggtttatact cagaagcact tctttgggtt tccctgcact 12781 atcctgatca tgtgctaggc ctaccttagg ctgattgttg ttcaaataaa cttaagtttc 12841 ctgtcaggtg atgtcatatg atttcatata tcaaggcaaa acatgttata tatgttaaac 12901 atttgtactt aatgtgaaag ttaggtcttt gtgggtttga tttttaattt tcaaaacctg 12961 agctaaataa gtcattttta catgtcttac atttggtgga attgtataat tgtggtttgc 13021 aggcaagact ctctgaccta gtaaccctac ctatagagca ctttgctggg tcacaagtct 13081 aggagtcaag catttcacct tgaagttgag acgttttgtt agtgtatact agtttatatg 13141 ttggaggaca tgtttatcca gaagatattc aggactattt ttgactgggc taaggaattg 13201 attctgatta gcactgttag tgagcattga gtggccttta ggcttgaatt ggagtcactt 13261 gtatatctca aataatgctg gcctttttta aaaagccctt gttctttatc accctgtttt 13321 ctacataatt tttgttcaaa gaaatacttg tttggatctc cttttgacaa caatagcatg 13381 ttttcaagcc atattttttt tccttttttt tttttttttt ggtttttcga gacagggttt 13441 ctctgtatag ccctggctgt cctggaactc actttgtaga ccaggctggc ctcgaactca 13501 gaaatccgcc tgcctctgcc tcctgagtgc cgggattaaa ggcgtgcacc accacgcctg 13561 gctaagttgg atattttgtt atataactat aaccaatact aactccactg ggtggatttt 13621 taattcagtc agtagtctta agtggtcttt attggccctt cattaaaatc tactgttcac 13681 tctaacagag gctgttggta ctagtggcac ttaagcaact tcctacggat atactagcag 13741 attaagggtc agggatagaa actagtctag cgttttgtat acctaccagc tttatactac 13801 cttgttctga tagaaatatt tcaggacatc tagcttatcg ataccgtcga cggtatcgat 13861 aagcttgata tcgaattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca 13921 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca 13981 cattccacat ccaccggtag gcgccgccgc cctgcagggc gatcgcggcc gcggtaccca 14041 gcttttgttc cctttagtga gggttaattg cgcgcttggc gtaatcatgg tcatagctgt 14101 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa 14161 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac 14221 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg 14281 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc 14341 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat 14401 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca 14461 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc 14521 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc 14581 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg 14641 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta 14701 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg 14761 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac 14821 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag 14881 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat 14941 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat 15001 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc 15061 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt 15121 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct 15181 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt 15241 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc 15301 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac 15361 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat 15421 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg 15481 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata 15541 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta 15601 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt 15661 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag 15721 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa 15781 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc 15841 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt 15901 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc 15961 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta 16021 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa 16081 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca 16141 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac 16201 aaataggggt tccgcgcaca tttccccgaa aagtgccacc taaattgtaa gcgttaatat 16261 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga 16321 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc 16381 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac 16441 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc 16501 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg 16561 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag 16621 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc 16681 gccgctacag ggcacgttcc attcgccatt caggctgcgc aactgttggg aagggcgatc 16741 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt 16801 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgagcg 16861 cgcgtaatac gactcactat agggcgaatt ggagct //
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