质粒名称: | MS2-P65-HSF1_GFP |
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出品公司: | 爱迪基因 |
目录编号: | 61423 |
存储实验室: | Feng Zhang |
载体骨架基本信息 |
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载体名称: | lenti(AMP) |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 哺乳细胞表达 |
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: | MS2(N55K)-P65-HSF1_2A_GFP |
物种来源: | --- |
插入大小: | -- |
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: | AgeI(未破坏) |
3' 克隆位点: | NotI (未破坏) |
5' 测序引物 | CMV-F |
3' 测序引物 | SV40pA-R |
文献引用方法 |
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材料和方法部分: | MS2-P65-HSF1_GFP was a gift from Feng Zhang (Addgene plasmid # 61423 ; http://n2t.net/addgene:61423 ; RRID:Addgene_61423) |
参考文献部分: | Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F. Nature. 2014 Dec 10. doi: 10.1038/nature14136. 10.1038/nature14136 PubMed 25494202 |
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载体序列
LOCUS MS2-P65-HSF1_GFP 11409 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Expresses the MS2-P65-HSF1 activator helper complex with a 2A GFP. 3rd generation lentiviral vector.. ACCESSION . VERSION . KEYWORDS MS2-P65-HSF1_GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 11409) AUTHORS Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F TITLE Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. JOURNAL Nature. 2014 Dec 10. doi: 10.1038/nature14136. PUBMED 25494202 REFERENCE 2 (bases 1 to 11409) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..11409 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(47..66) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 238..617 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 619..817 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 772..792 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" LTR 835..1015 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1062..1187 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1680..1913 /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 2440..2557 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2682..3859 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 2913..3850 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 3807..3827 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" regulatory 3866..3875 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 3875..4261 /codon_start=1 /product="bacteriophage MS2 coat protein" /label=MS2-N55K /note="binds with high affinity to a specific stem-loop structure in the viral RNA (Lim et al., 1994)" /translation="ASNFTQFVLVDNGGTGDVTVAPSNFANGVAEWISSNSRSQAYKVT CSVRQSSAQKRKYTIKVEVPKVATQTVGGVELPVAAWRSYLNMELTIPIFATNSDCELI VKAMQGLLKDGNPIPSAIAANSGIY" CDS 4316..4336 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 4352..4894 /codon_start=1 /product="C-terminal portion of the p65 subunit of mouse NF-kappa-B" /label=p65 /translation="PSGQISNQALALAPSSAPVLAQTMVPSSAMVPLAQPPAPAPVLTP GPPQSLSAPVPKSTQAGEGTLSEALLHLQFDADEDLGALLGNSTDPGVFTDLASVDNSE FQQLLNQGVSMSHSTAEPMLMEYPEAITRLVTGSQRPPDPAPTPLGTSGLPNGLSGDED FSSIADMDFSALLSQISS" CDS 4919..5290 /codon_start=1 /product="C-terminal activation domain from the human heat shock transcription factor HSF1" /label=HSF1 /translation="GFSVDTSALLDLFSPSVTVPDMSLPDLDSSLASIQELLSPQEPPR PPEAENSSPDSGKQLVHYTAQPLFLLDPGSVDTGSNDLPVLFELGEGSYFSEGDGFAED PTISLLTGSEPPKAKDPTVS" CDS 5306..5359 /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 5360..6076 /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE FVTAAGITLGMDELYK" primer_bind complement(5402..5423) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(5663..5682) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 6010..6031 /label=EGFP-C /note="EGFP, forward primer" misc_feature 6101..6689 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6154..6174) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6572..6583) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(6692..6708) /label=KS primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6693..6709) /label=pBluescriptKS /note="For pBluescript vector" LTR 7214..7394 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" primer_bind complement(7420..7437) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 7426..7650 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 7696..8124 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7783..7802) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7993..8014 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(8133..8153) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 8138..8467 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 8318..8453 /label=SV40 ori /note="SV40 origin of replication" primer_bind 8380..8399 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" promoter 8515..8562 /label=EM7 promoter /note="synthetic bacterial promoter " CDS 8581..8955 /codon_start=1 /gene="Sh ble from Streptoalloteichus hindustanus" /product="antibiotic-binding protein" /label=BleoR /note="confers resistance to bleomycin, phleomycin, and Zeocin(TM)" /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR EFALRDPAGNCVHFVAEEQD" polyA_signal 9085..9206 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9122..9141) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9176..9195 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(9255..9271) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9255..9271) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(9268..9290) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 9279..9295 /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(9303..9333) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 9348..9369 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(9486..9503) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(9657..10245) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(9737..9756) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(10416..11276) /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 11039..11058 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(11277..11381) /gene="bla" /label=AmpR promoter ORIGIN 1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg 61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt 121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc 181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac 241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat 301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg 361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt 421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag 481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc 541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag 601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt 661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc 721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg 781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct 841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt 901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac 961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc 1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc 1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa 1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg 1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata 1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc 1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga 1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc 1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca 1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg 1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga 1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata 1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg 1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg 1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag 1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt 1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt 1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt 2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag 2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata 2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta 2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta 2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa 2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt 2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat 2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa 2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag 2581 agatccagtt tggttaatta atgcaaagat ggataaagtt ttaaacagag aggaatcttt 2641 gcagctaatg gaccttctag gtcttgaaag gagtgggaat tggctccggt gcccgtcagt 2701 gggcagagcg cacatcgccc acagtccccg agaagttggg gggaggggtc ggcaattgaa 2761 ccggtgccta gagaaggtgg cgcggggtaa actgggaaag tgatgtcgtg tactggctcc 2821 gcctttttcc cgagggtggg ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc 2881 tttttcgcaa cgggtttgcc gccagaacac aggtaagtgc cgtgtgtggt tcccgcgggc 2941 ctggcctctt tacgggttat ggcccttgcg tgccttgaat tacttccact ggctgcagta 3001 cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga ggccttgcgc 3061 ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct ggggccgccg 3121 cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt ctctagccat 3181 ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc ttgtaaatgc 3241 gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg acggggcccg 3301 tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac cgagaatcgg 3361 acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc cgccgtgtat 3421 cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag cggaaagatg 3481 gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct cgggagagcg 3541 ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg tcgcttcatg 3601 tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga gcttttggag 3661 tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc acactgagtg 3721 ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg aatttgccct 3781 ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa gtttttttct 3841 tccatttcag gtgtcgtgac gtacggccac catggcttca aactttactc agttcgtgct 3901 cgtggacaat ggtgggacag gggatgtgac agtggctcct tctaatttcg ctaatggggt 3961 ggcagagtgg atcagctcca actcacggag ccaggcctac aaggtgacat gcagcgtcag 4021 gcagtctagt gcccagaaga gaaagtatac catcaaggtg gaggtcccca aagtggctac 4081 ccagacagtg ggcggagtcg aactgcctgt cgccgcttgg aggtcctacc tgaacatgga 4141 gctcactatc ccaattttcg ctaccaattc tgactgtgaa ctcatcgtga aggcaatgca 4201 ggggctcctc aaagacggta atcctatccc ttccgccatc gccgctaact caggtatcta 4261 cagcgctgga ggaggtggaa gcggaggagg aggaagcgga ggaggaggta gcggacctaa 4321 gaaaaagagg aaggtggcgg ccgctggatc cccttcaggg cagatcagca accaggccct 4381 ggctctggcc cctagctccg ctccagtgct ggcccagact atggtgccct ctagtgctat 4441 ggtgcctctg gcccagccac ctgctccagc ccctgtgctg accccaggac caccccagtc 4501 actgagcgct ccagtgccca agtctacaca ggccggcgag gggactctga gtgaagctct 4561 gctgcacctg cagttcgacg ctgatgagga cctgggagct ctgctgggga acagcaccga 4621 tcccggagtg ttcacagatc tggcctccgt ggacaactct gagtttcagc agctgctgaa 4681 tcagggcgtg tccatgtctc atagtacagc cgaaccaatg ctgatggagt accccgaagc 4741 cattacccgg ctggtgaccg gcagccagcg gccccccgac cccgctccaa ctcccctggg 4801 aaccagcggc ctgcctaatg ggctgtccgg agatgaagac ttctcaagca tcgctgatat 4861 ggactttagt gccctgctgt cacagatttc ctctagtggg cagggaggag gtggaagcgg 4921 cttcagcgtg gacaccagtg ccctgctgga cctgttcagc ccctcggtga ccgtgcccga 4981 catgagcctg cctgaccttg acagcagcct ggccagtatc caagagctcc tgtctcccca 5041 ggagcccccc aggcctcccg aggcagagaa cagcagcccg gattcaggga agcagctggt 5101 gcactacaca gcgcagccgc tgttcctgct ggaccccggc tccgtggaca ccgggagcaa 5161 cgacctgccg gtgctgtttg agctgggaga gggctcctac ttctccgaag gggacggctt 5221 cgccgaggac cccaccatct ccctgctgac aggctcggag cctcccaaag ccaaggaccc 5281 cactgtctcc gctagcggca gtggagaggg cagaggaagt ctgctaacat gcggtgacgt 5341 cgaggagaat cctggcccag tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat 5401 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga 5461 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc 5521 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta 5581 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca 5641 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt 5701 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg 5761 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc 5821 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg 5881 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct 5941 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa 6001 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga 6061 cgagctgtac aagtaagaat tcgatatcaa gcttatcgat aatcaacctc tggattacaa 6121 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata 6181 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc 6241 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg 6301 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac 6361 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat 6421 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt 6481 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat 6541 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc 6601 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag 6661 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa 6721 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga 6781 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat 6841 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg 6901 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg 6961 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt 7021 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg 7081 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga 7141 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca 7201 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta 7261 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg 7321 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg 7381 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt 7441 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc 7501 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt 7561 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca 7621 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct 7681 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta 7741 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc 7801 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt 7861 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg 7921 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca 7981 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct 8041 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga 8101 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa 8161 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac 8221 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa 8281 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag 8341 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc 8401 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt 8461 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac 8521 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc 8581 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc 8641 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt 8701 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac 8761 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag 8821 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag 8881 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc 8941 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg 9001 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc 9061 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa 9121 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt 9181 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc 9241 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca 9301 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa 9361 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag 9421 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc 9481 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct 9541 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg 9601 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc 9661 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga 9721 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct 9781 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg 9841 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag 9901 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat 9961 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac 10021 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac 10081 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc 10141 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt 10201 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc 10261 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg 10321 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca 10381 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca 10441 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag 10501 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac 10561 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc 10621 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct 10681 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc 10741 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg 10801 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc 10861 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat 10921 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag 10981 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat 11041 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg 11101 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca 11161 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga 11221 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc 11281 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata 11341 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg 11401 ccacctgac //