质粒名称: | Ai6 |
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出品公司: | Addgene |
目录编号: | 22798 |
存储实验室: | Hongkui Zeng |
载体骨架基本信息 |
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载体名称: | Rosa26 Targeting Vector |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 9862 bp |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | Mouse Targeting |
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 |
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插入基因名称: | CAG-Floxed ZsGreen |
物种来源: | n/a |
插入大小: | 6319 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: | PacI(未破坏) |
3' 克隆位点: | AscI (未破坏) |
5' 测序引物 | n/a |
3' 测序引物 | n/a |
文献引用方法 |
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材料和方法部分: | Ai6 was a gift from Hongkui Zeng (Addgene plasmid # 22798 ; http://n2t.net/addgene:22798 ; RRID:Addgene_22798) |
参考文献部分: | A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, Ng LL, Palmiter RD, Hawrylycz MJ, Jones AR, Lein ES, Zeng H. Nat Neurosci. 2010 Jan . 13(1):133-40. 10.1038/nn.2467 PubMed 20023653 |
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 Ai6 16015 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Ai6 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 16015) AUTHORS Madisen L, Zwingman TA, Sunkin SM, Oh SW, Zariwala HA, Gu H, Ng LL, Palmiter RD, Hawrylycz MJ, Jones AR, Lein ES, Zeng H TITLE A robust and high-throughput Cre reporting and characterization system for the whole mouse brain. JOURNAL Nat Neurosci. 2010 Jan . 13(1):133-40. PUBMED 20023653 REFERENCE 2 (bases 1 to 16015) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..16015 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 947..1250 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1252..1529 /label=chicken beta-actin promoter intron 1530..2546 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 2470..2492 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 2554..2573 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" protein_bind 2622..2655 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" protein_bind complement(2679..2712) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." polyA_signal 2879..3013 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" polyA_signal 3133..3267 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" polyA_signal 3387..3521 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" protein_bind complement(3550..3583) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." regulatory 3644..3653 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3650..4345 /codon_start=1 /product="Zoanthus green fluorescent protein" /label=ZsGreen1 /note="mammalian codon-optimized" /translation="MAQSKHGLTKEMTMKYRMEGCVDGHKFVITGEGIGYPFKGKQAIN LCVVEGGPLPFAEDILSAAFMYGNRVFTEYPQDIVDYFKNSCPAGYTWDRSFLFEDGAV CICNADITVSVEENCMYHESKFYGVNFPADGPVMKKMTDNWEPSCEKIIPVPKQGILKG DVSMYLLLKDGGRLRCQFDTVYKAKSVPRKMPDWHFIQHKLTREDRSDAKNQKWHLTEH AIASGSALP" misc_feature 4393..4981 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(4446..4466) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(4864..4875) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(4984..5000) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4985..5001) /label=pBluescriptKS /note="For pBluescript vector" polyA_signal 5008..5215 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" protein_bind 5223..5256 /label=attB /bound_moiety="phage phi-C31 integrase" /note="minimal attB site for the phi-C31 integrase (Groth et al., 2000)" promoter 5278..5777 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" protein_bind 5797..5830 /label=FRT (minimal) /bound_moiety="FLP recombinase from the Saccharomyces cerevisiae 2u plasmid" /note="supports FLP-mediated excision but not integration (Turan and Bode, 2011)" regulatory 5831..5840 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5837..6640 /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 EHQGLAPAELFARLKARMPDGDDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5900..5919) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 6510..6529 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6650..7111 /label=PGK poly(A) signal /note="mouse phosphoglycerate kinase 1 polyadenylation signal" misc_feature 7177..11517 /label=Rosa26 right arm /note="right homology arm from intron 1 of the mouse Rosa26 gene (Soriano, 1999)" promoter 11567..12066 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" CDS 12093..12665 /codon_start=1 /gene="Corynebacterium diphtheriae tox" /product="diphtheria toxin fragment A" /label=DTA /translation="DDVVDSSKSFVMENFSSYHGTKPGYVDSIQKGIQKPKSGTQGNYD DDWKGFYSTDNKYDAAGYSVDNENPLSGKAGGVVKVTYPGLTKVLALKVDNAETIKKEL GLSLTEPLMEQVGTEEFIKRFGDGASRVVLSLPFAEGSSSVEYINNWEQAKALSVELEI NFETRGKRGQDAMYEYMAQACAGNRVRR" intron 12742..12807 /label=small t intron /note="SV40 (simian virus 40) small t antigen intron" polyA_signal 12881..13105 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" primer_bind complement(13167..13187) /label=T3 /note="T3 promoter, forward primer" promoter complement(13167..13185) /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" primer_bind complement(13206..13222) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(13206..13222) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(13219..13241) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 13230..13246 /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(13254..13284) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 13299..13320 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(13437..13454) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(13608..14196) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(13688..13707) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(14367..15227) /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 14990..15009 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(15228..15332) /gene="bla" /label=AmpR promoter rep_origin 15359..15814 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(15446..15465) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 15656..15677 /label=F1ori-F /note="F1 origin, forward primer" primer_bind 15940..15962 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 15954..15971 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 15955..15971 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 15981..16000 /label=T7 /note="T7 promoter, forward primer" promoter 15981..15999 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 ccgcggcagg ccctccgagc gtggtggagc cgttctgtga gacagccggg tacgagtcgt 61 gacgctggaa ggggcaagcg ggtggtgggc aggaatgcgg tccgccctgc agcaaccgga 121 gggggaggga gaagggagcg gaaaagtctc caccggacgc ggccatggct cggggggggg 181 gggggggagg gagggtcagc gaaagtagct cgcgcgcgag cggccgccca ccctcccctt 241 cctctggggg agtcgtttta cccgccgccg gccgggcctc gtcgtctgat tggctctcgg 301 ggcccagaaa actggccctt gccattggct cgtgttcgtg caagttgagt ccatccgccg 361 gccagcgggg gcggcgagga ggcgctccca ggttccggcc ctcccctcgg ccccgcgccg 421 cagagtctgg ccgcgcgccc ctgcgcaacg tggcaggaag cgcgcgctgg gggcggggac 481 gggcagtagg gctgagcggc tgcggggcgg gtgcaagcac gtttccgact tgagttgcct 541 caagaggggc gtgctgagcc agacctccat cgcgcactcc ggggagtgga gggaaggagc 601 gagggctcag ttgggctgtt ttggaggcag gaagcacttg ctctcccaaa gtcgctctga 661 gttgttatca gtaagggagc tgcagtggag taggcgggga gaaggccgca cccttctccg 721 gaggggggag gggagtgttg caataccttt ctgggagttc tctgctgcct cctggcttct 781 gaggaccgcc ctgggcctgg gagaatccct tccccctctt ccctcgtgat ctgcaactcc 841 agtctttcta gccttaatta accgtttaaa caattctgca ggaatctagt tattaatagt 901 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta 961 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga 1021 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt 1081 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta 1141 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg 1201 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtcgaggtga 1261 gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca attttgtatt 1321 tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg gggcgcgcgc 1381 caggcggggc ggggcggggc gaggggcggg gcggggcgag gcggagaggt gcggcggcag 1441 ccaatcagag cggcgcgctc cgaaagtttc cttttatggc gaggcggcgg cggcggcggc 1501 cctataaaaa gcgaagcgcg cggcgggcgg gagtcgctgc gcgctgcctt cgccccgtgc 1561 cccgctccgc cgccgcctcg cgccgcccgc cccggctctg actgaccgcg ttactcccac 1621 aggtgagcgg gcgggacggc ccttctcctc cgggctgtaa ttagcgcttg gtttaatgac 1681 ggcttgtttc ttttctgtgg ctgcgtgaaa gccttgaggg gctccgggag ggccctttgt 1741 gcggggggag cggctcgggg ggtgcgtgcg tgtgtgtgtg cgtggggagc gccgcgtgcg 1801 gctccgcgct gcccggcggc tgtgagcgct gcgggcgcgg cgcggggctt tgtgcgctcc 1861 gcagtgtgcg cgaggggagc gcggccgggg gcggtgcccc gcggtgcggg gggggctgcg 1921 aggggaacaa aggctgcgtg cggggtgtgt gcgtgggggg gtgagcaggg ggtgtgggcg 1981 cgtcggtcgg gctgcaaccc cccctgcacc cccctccccg agttgctgag cacggcccgg 2041 cttcgggtgc ggggctccgt acggggcgtg gcgcggggct cgccgtgccg ggcggggggt 2101 ggcggcaggt gggggtgccg ggcggggcgg ggccgcctcg ggccggggag ggctcggggg 2161 aggggcgcgg cggcccccgg agcgccggcg gctgtcgagg cgcggcgagc cgcagccatt 2221 gccttttatg gtaatcgtgc gagagggcgc agggacttcc tttgtcccaa atctgtgcgg 2281 agccgaaatc tgggaggcgc cgccgcaccc cctctagcgg gcgcggggcg aagcggtgcg 2341 gcgccggcag gaaggaaatg ggcggggagg gccttcgtgc gtcgccgcgc cgccgtcccc 2401 ttctccctct ccagcctcgg ggctgtccgc ggggggacgg ctgccttcgg gggggacggg 2461 gcagggcggg gttcggcttc tggcgtgtga ccggcggctc tagagcctct gctaaccatg 2521 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc 2581 atcattttgg caaagaattg atttgatacc gcgggcccta agaagttcct attctctaga 2641 aagtatagga acttcgtcga catttaaatc atttaaatat aacttcgtat aatgtatgct 2701 atacgaagtt attcgcgatg aataaatgaa agcttgcaga tctgcgactc tagaggatct 2761 gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 2821 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 2881 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 2941 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 3001 tcatgtctgg atctgcgact ctagaggatc ataatcagcc ataccacatt tgtagaggtt 3061 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca 3121 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc 3181 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc 3241 atcaatgtat cttatcatgt ctggatctgc gactctagag gatcataatc agccatacca 3301 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac 3361 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat 3421 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg 3481 gtttgtccaa actcatcaat gtatcttatc atgtctggat ccccatcaag ctgatccgga 3541 acccttaata taacttcgta taatgtatgc tatacgaagt tattaggtcc ctcgacctgc 3601 agcccaagct agatcgaatt cggccggccg atatcggcgc gccgccacca tggcccagtc 3661 caagcacggc ctgaccaagg agatgaccat gaagtaccgc atggagggct gcgtggacgg 3721 ccacaagttc gtgatcaccg gcgagggcat cggctacccc ttcaagggca agcaggccat 3781 caacctgtgc gtggtggagg gcggcccctt gcccttcgcc gaggacatct tgtccgccgc 3841 cttcatgtac ggcaaccgcg tgttcaccga gtacccccag gacatcgtcg actacttcaa 3901 gaactcctgc cccgccggct acacctggga ccgctccttc ctgttcgagg acggcgccgt 3961 gtgcatctgc aacgccgaca tcaccgtgag cgtggaggag aactgcatgt accacgagtc 4021 caagttctac ggcgtgaact tccccgccga cggccccgtg atgaagaaga tgaccgacaa 4081 ctgggagccc tcctgcgaga agatcatccc cgtgcccaag cagggcatct tgaagggcga 4141 cgtgagcatg tacctgctgc tgaaggacgg tggccgcttg cgctgccagt tcgacaccgt 4201 gtacaaggcc aagtccgtgc cccgcaagat gcccgactgg cacttcatcc agcacaagct 4261 gacccgcgag gaccgcagcg acgccaagaa ccagaagtgg cacctgaccg agcacgccat 4321 cgcctccggc tccgccttgc cctgataaga tatcggccgg ccctgcagga attcgatatc 4381 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt 4441 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct 4501 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt 4561 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac 4621 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct 4681 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca 4741 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt 4801 ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc 4861 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct 4921 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg 4981 catcgatacc gtcgacctcg acctcgactg tgccttctag ttgccagcca tctgttgttt 5041 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat 5101 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg 5161 tggggcagga cagcaagggg gaggattggg aagacaatgg caggcatgct ggggaactag 5221 tggtgccagg gcgtgccctt gggctccccg ggcgcggcgg ccgcatcgaa ttctaccggg 5281 taggggaggc gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca 5341 cttggcgcta cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc 5401 aaccggctcc gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga 5461 agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc 5521 tcactagtct cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg 5581 gcagcggcca atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg 5641 ggtccggggg cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg 5701 gaggcccggc attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct 5761 catctccggg cctttcgacc tgcaatcgcc gctagcgaag ttcctattct ctagaaagta 5821 taggaacttc gccaccatgg gatcggccat tgaacaagat ggattgcacg caggttctcc 5881 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc 5941 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga 6001 cctgtccggt gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac 6061 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct 6121 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa 6181 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc 6241 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct 6301 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc 6361 caggctcaag gcgcgcatgc ccgacggcga tgatctcgtc gtgacccatg gcgatgcctg 6421 cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct 6481 gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct 6541 tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca 6601 gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga ggggatccgc tgtaagtctg 6661 cagaaattga tgatctatta aacaataaag atgtccacta aaatggaagt ttttcctgtc 6721 atactttgtt aagaagggtg agaacagagt acctacattt tgaatggaag gattggagct 6781 acgggggtgg gggtggggtg ggattagata aatgcctgct ctttactgaa ggctctttac 6841 tattgcttta tgataatgtt tcatagttgg atatcataat ttaaacaagc aaaaccaaat 6901 taagggccag ctcattcctc ccactcatga tctatagatc tatagatctc tcgtgggatc 6961 attgtttttc tcttgattcc cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca 7021 gtttcatagc ctgaagaacg agatcagcag cctctgttcc acatacactt cattctcagt 7081 attgttttgc caagttctaa ttccatcaga aagcttgcag aagatctccc caactggggt 7141 aacctttgag ttctctcagt tgggggggcg cgccggctag aagatgggcg ggagtcttct 7201 gggcaggctt aaaggctaac ctggtgtgtg ggcgttgtcc tgcaggggaa ttgaacaggt 7261 gtaaaattgg agggacaaga cttcccacag attttcggtt ttgtcgggaa gttttttaat 7321 aggggcaaat aaggaaaatg ggaggatagg tagtcatctg gggttttatg cagcaaaact 7381 acaggttatt attgcttgtg atccgcctcg gagtattttc catcgaggta gattaaagac 7441 atgctcaccc gagttttata ctctcctgct tgagatcctt actacagtat gaaattacag 7501 tgtcgcgagt tagactatgt aagcagaatt ttaatcattt ttaaagagcc cagtacttca 7561 tatccatttc tcccgctcct tctgcagcct tatcaaaagg tattttagaa cactcatttt 7621 agccccattt tcatttatta tactggctta tccaacccct agacagagca ttggcatttt 7681 ccctttcctg atcttagaag tctgatgact catgaaacca gacagattag ttacatacac 7741 cacaaatcga ggctgtagct ggggcctcaa cactgcagtt cttttataac tccttagtac 7801 actttttgtt gatcctttgc cttgatcctt aattttcagt gtctatcacc tctcccgtca 7861 ggtggtgttc cacatttggg cctattctca gtccagggag ttttacaaca atagatgtat 7921 tgagaatcca acctaaagct taactttcca ctcccatgaa tgcctctctc ctttttctcc 7981 atttataaac tgagctatta accattaatg gtttccaggt ggatgtctcc tcccccaata 8041 ttacctgatg tatcttacat attgccaggc tgatatttta agacattaaa aggtatattt 8101 cattattgag ccacatggta ttgattactg cttactaaaa ttttgtcatt gtacacatct 8161 gtaaaaggtg gttccttttg gaatgcaaag ttcaggtgtt tgttgtcttt cctgacctaa 8221 ggtcttgtga gcttgtattt tttctattta agcagtgctt tctcttggac tggcttgact 8281 catggcattc tacacgttat tgctggtcta aatgtgattt tgccaagctt cttcaggacc 8341 tataattttg cttgacttgt agccaaacac aagtaaaatg attaagcaac aaatgtattt 8401 gtgaagcttg gtttttaggt tgttgtgttg tgtgtgcttg tgctctataa taatactatc 8461 caggggctgg agaggtggct cggagttcaa gagcacagac tgctcttcca gaagtcctga 8521 gttcaattcc cagcaaccac atggtggctc acaaccatct gtaatgggat ctgatgccct 8581 cttctggtgt gtctgaagac cacaagtgta ttcacattaa ataaataaat cctccttctt 8641 cttctttttt ttttttttaa agagaatact gtctccagta gaatttactg aagtaatgaa 8701 atactttgtg tttgttccaa tatggtagcc aataatcaaa ttactcttta agcactggaa 8761 atgttaccaa ggaactaatt tttatttgaa gtgtaactgt ggacagagga gccataactg 8821 cagacttgtg ggatacagaa gaccaatgca gactttaatg tcttttctct tacactaagc 8881 aataaagaaa taaaaattga acttctagta tcctatttgt ttaaactgct agctttactt 8941 aacttttgtg cttcatctat acaaagctga aagctaagtc tgcagccatt actaaacatg 9001 aaagcaagta atgataattt tggatttcaa aaatgtaggg ccagagttta gccagccagt 9061 ggtggtgctt gcctttatgc ctttaatccc agcactctgg aggcagagac aggcagatct 9121 ctgagtttga gcccagcctg gtctacacat caagttctat ctaggatagc caggaataca 9181 cacagaaacc ctgttgggga ggggggctct gagatttcat aaaattataa ttgaagcatt 9241 ccctaatgag ccactatgga tgtggctaaa tccgtctacc tttctgatga gatttgggta 9301 ttattttttc tgtctctgct gttggttggg tcttttgaca ctgtgggctt tctttaaagc 9361 ctccttcctg ccatgtggtc tcttgtttgc tactaacttc ccatggctta aatggcatgg 9421 ctttttgcct tctaagggca gctgctgaga tttgcagcct gatttccagg gtggggttgg 9481 gaaatctttc aaacactaaa attgtccttt aatttttttt ttaaaaaatg ggttatataa 9541 taaacctcat aaaatagtta tgaggagtga ggtggactaa tattaaatga gtccctcccc 9601 tataaaagag ctattaaggc tttttgtctt atacttaact ttttttttaa atgtggtatc 9661 tttagaacca agggtcttag agttttagta tacagaaact gttgcatcgc ttaatcagat 9721 tttctagttt caaatccaga gaatccaaat tcttcacagc caaagtcaaa ttaagaattt 9781 ctgactttta atgttaattt gcttactgtg aatataaaaa tgatagcttt tcctgaggca 9841 gggtctcact atgtatctct gcctgatctg caacaagata tgtagactaa agttctgcct 9901 gcttttgtct cctgaatact aaggttaaaa tgtagtaata cttttggaac ttgcaggtca 9961 gattctttta taggggacac actaagggag cttgggtgat agttggtaaa atgtgtttca 10021 agtgatgaaa acttgaatta ttatcaccgc aacctacttt ttaaaaaaaa aagccaggcc 10081 tgttagagca tgcttaaggg atccctagga cttgctgagc acacaagagt agttacttgg 10141 caggctcctg gtgagagcat atttcaaaaa acaaggcaga caaccaagaa actacagtta 10201 aggttacctg tctttaaacc atctgcatat acacagggat attaaaatat tccaaataat 10261 atttcattca agttttcccc catcaaattg ggacatggat ttctccggtg aataggcaga 10321 gttggaaact aaacaaatgt tggttttgtg atttgtgaaa ttgttttcaa gtgatagtta 10381 aagcccatga gatacagaac aaagctgcta tttcgaggtc tcttggttta tactcagaag 10441 cacttctttg ggtttccctg cactatcctg atcatgtgct aggcctacct taggctgatt 10501 gttgttcaaa taaacttaag tttcctgtca ggtgatgtca tatgatttca tatatcaagg 10561 caaaacatgt tatatatgtt aaacatttgt acttaatgtg aaagttaggt ctttgtgggt 10621 ttgattttta attttcaaaa cctgagctaa ataagtcatt tttacatgtc ttacatttgg 10681 tggaattgta taattgtggt ttgcaggcaa gactctctga cctagtaacc ctacctatag 10741 agcactttgc tgggtcacaa gtctaggagt caagcatttc accttgaagt tgagacgttt 10801 tgttagtgta tactagttta tatgttggag gacatgttta tccagaagat attcaggact 10861 atttttgact gggctaagga attgattctg attagcactg ttagtgagca ttgagtggcc 10921 tttaggcttg aattggagtc acttgtatat ctcaaataat gctggccttt tttaaaaagc 10981 ccttgttctt tatcaccctg ttttctacat aatttttgtt caaagaaata cttgtttgga 11041 tctccttttg acaacaatag catgttttca agccatattt tttttccttt tttttttttt 11101 ttttggtttt tcgagacagg gtttctctgt atagccctgg ctgtcctgga actcactttg 11161 tagaccaggc tggcctcgaa ctcagaaatc cgcctgcctc tgcctcctga gtgccgggat 11221 taaaggcgtg caccaccacg cctggctaag ttggatattt tgttatataa ctataaccaa 11281 tactaactcc actgggtgga tttttaattc agtcagtagt cttaagtggt ctttattggc 11341 ccttcattaa aatctactgt tcactctaac agaggctgtt ggtactagtg gcacttaagc 11401 aacttcctac ggatatacta gcagattaag ggtcagggat agaaactagt ctagcgtttt 11461 gtatacctac cagctttata ctaccttgtt ctgatagaaa tatttcagga catctagctt 11521 atcgataccg tcgacggtat cgataagctt gatatcgaat tctaccgggt aggggaggcg 11581 cttttcccaa ggcagtctgg agcatgcgct ttagcagccc cgctgggcac ttggcgctac 11641 acaagtggcc tctggcctcg cacacattcc acatccaccg gtaggcgcca accggctccg 11701 ttctttggtg gccccttcgc gccaccttct actcctcccc tagtcaggaa gttccccccc 11761 gccccgcagc tcgcgtcgtg caggacgtga caaatggaag tagcacgtct cactagtctc 11821 gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag gcctttgggg cagcggccaa 11881 tagcagcttt gctccttcgc tttctgggct cagaggctgg gaaggggtgg gtccgggggc 11941 gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa ggtcctccgg aggcccggca 12001 ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct cctcttcctc atctccgggc 12061 ctttcgacct gcaggtcctc gccatggatc ctgatgatgt tgttgattct tctaaatctt 12121 ttgtgatgga aaacttttct tcgtaccacg ggactaaacc tggttatgta gattccattc 12181 aaaaaggtat acaaaagcca aaatctggta cacaaggaaa ttatgacgat gattggaaag 12241 ggttttatag taccgacaat aaatacgacg ctgcgggata ctctgtagat aatgaaaacc 12301 cgctctctgg aaaagctgga ggcgtggtca aagtgacgta tccaggactg acgaaggttc 12361 tcgcactaaa agtggataat gccgaaacta ttaagaaaga gttaggttta agtctcactg 12421 aaccgttgat ggagcaagtc ggaacggaag agtttatcaa aaggttcggt gatggtgctt 12481 cgcgtgtagt gctcagcctt cccttcgctg aggggagttc tagcgttgaa tatattaata 12541 actgggaaca ggcgaaagcg ttaagcgtag aacttgagat taattttgaa acccgtggaa 12601 aacgtggcca agatgcgatg tatgagtata tggctcaagc ctgtgcagga aatcgtgtca 12661 ggcgatctct ttgtgaagga accttacttc tgtggtgtga cataattgga caaactacct 12721 acagagattt aaagctctaa ggtaaatata aaatttttaa gtgtataatg tgttaaacta 12781 ctgattctaa ttgtttgtgt attttagatt ccaacctatg gaactgatga atgggagcag 12841 tggtggaatg cagatcctag agctcgctga tcagcctcga ctgtgccttc tagttgccag 12901 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact 12961 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt 13021 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat 13081 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgagg 13141 gggggcccgg tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa 13201 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata 13261 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta 13321 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa 13381 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg 13441 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag 13501 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa 13561 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc 13621 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca 13681 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg 13741 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct 13801 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt 13861 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag 13921 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc 13981 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac 14041 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga 14101 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc 14161 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg 14221 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca 14281 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt 14341 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca 14401 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg 14461 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca 14521 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt 14581 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt 14641 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca 14701 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca 14761 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga 14821 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact 14881 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga 14941 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg 15001 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc 15061 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga 15121 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat 15181 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt 15241 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt 15301 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac 15361 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 15421 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 15481 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 15541 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 15601 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 15661 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 15721 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 15781 gcgaatttta acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact 15841 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat 15901 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa 15961 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattggag ctcca //