质粒名称: | Apple-53BP1trunc |
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出品公司: | Addgene |
目录编号: | 69531 |
存储实验室: | Ralph Weissleder |
载体骨架基本信息 |
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载体名称: | pLVX |
载体出品公司: | Clontech |
空载体大小: | 8003 bp |
完整质粒大小: | 10184 bp |
载体修饰: | -- |
载体类型: | 哺乳细胞表达,慢病毒载体 |
筛选标记: | 嘌呤霉素 |
载体抗性: | 氨苄霉素100 μg/mL |
生长温度: | 37 ℃ |
宿主菌: | NEB Stable |
拷贝数: | -- |
插入基因信息 |
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插入基因名称: | 53BP1,TP53BP1 |
物种来源: | H. sapiens (human) |
插入大小: | 1473 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子: | CMV |
标签/融合蛋白: | Apple fluorescent protein(插入物上的N端) |
克隆方法: | 限制性内切酶 |
5’克隆位点: | BsrGI (未破坏) |
3’克隆位点: | XbaI(克隆期间销毁) |
5’测序引物 | CMV-F |
3’测序引物 | -- |
文献引用方法 |
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材料和方法部分: | Apple-53BP1trunc was a gift from Ralph Weissleder (Addgene plasmid # 69531 ; http://n2t.net/addgene:69531 ; RRID:Addgene_69531) |
参考文献部分: | Single cell resolution in vivo imaging of DNA damage following PARP inhibition. Yang KS, Kohler RH, Landon M, Giedt R, Weissleder R. Sci Rep. 2015 May 18;5:10129. doi: 10.1038/srep10129. 10.1038/srep10129 PubMed 25984718 |
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Apple53BP1trunc载体基本信息
Apple53BP1trunc载体图谱质粒图谱和多克隆位点信息
Apple53BP1trunc载体简介
Apple53BP1trunc载体序列
LOCUS Apple-53BP1trunc 10961 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent reporter for DNA damage. ACCESSION . VERSION . KEYWORDS Apple-53BP1trunc SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10961) AUTHORS Yang KS, Kohler RH, Landon M, Giedt R, Weissleder R TITLE Single cell resolution in vivo imaging of DNA damage following PARP inhibition. JOURNAL Sci Rep. 2015 May 18;5:10129. doi: 10.1038/srep10129. PUBMED 25984718 REFERENCE 2 (bases 1 to 10961) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10961 /organism="synthetic DNA construct" /mol_type="other DNA" LTR 1..634 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" misc_feature 681..806 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1303..1536 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 1721..1765 /codon_start=1 /product="antigenic peptide corresponding to amino acids 655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik et al., 2013)" /label=gp41 peptide /note="recognized by the 2H10 single-chain llama nanobody" /translation="KNEQELLELDKWASL" misc_feature 2028..2143 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1 (lacking the first T)" enhancer 2201..2504 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 2505..2708 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 2659..2679 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 2705..2729 /label=LNCX /note="Human CMV promoter, forward primer" CDS 2820..3527 /codon_start=1 /product="photostable monomeric derivative of DsRed (Shaner et al., 2008)" /label=mApple /note="mammalian codon-optimized" /translation="MVSKGEENNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEA FQTAKLKVTKGGPLPFAWDILSPQFMYGSKVYIKHPADIPDYFKLSFPEGFRWERVMNF EDGGIIHVNQDSSLQDGVFIYKVKLRGTNFPSDGPVMQKKTMGWEASEERMYPEDGALK SEIKKRLKLKDGGHYAAEVKTTYKAKKPVQLPGAYIVDIKLDIVSHNEDYTIVEQYERA EGRHSTGGMDELYK" primer_bind 3233..3252 /label=mCherry-F /note="mCherry, forward primer" promoter 5019..5518 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(5054..5077) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 5451..5470 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 5539..6138 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(5539..5558) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 6035..6055 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 6152..6740 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(6205..6225) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(6623..6634) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6947..7580 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7709..7725) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7709..7725) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7722..7744) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7733..7749 /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(7757..7787) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7802..7823 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7940..7957) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(8111..8699) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(8191..8210) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8870..9730) /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 9493..9512 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" mobile_element 9741..10508 /mobile_element_type="insertion sequence:IS1" /note="prokaryotic transposable element" promoter complement(10509..10612) /gene="bla" /label=AmpR promoter polyA_signal 10660..10794 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(10697..10716) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 10751..10770 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" ORIGIN 1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca 61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac 121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca 181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg 241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag 301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg 361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat 421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga 481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct 541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc 601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag 661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg 721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga 781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg 841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg 901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct 961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat 1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca 1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc 1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag 1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc 1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg 1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc 1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc 1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct 1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa 1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca 1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt 1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt 1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta 1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt 1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct 1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga 1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag 2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac 2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg 2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa 2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg 2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt 2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg 2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc 2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc 2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca 2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta 2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa 2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc 2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccgggata gctagcacca 2821 tggtgagcaa gggcgaggag aataacatgg ccatcatcaa ggagttcatg cgcttcaagg 2881 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc 2941 gcccctacga ggcctttcag accgctaagc tgaaggtgac caagggtggc cccctgccct 3001 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggtctac attaagcacc 3061 cagccgacat ccccgactac ttcaagctgt ccttccccga gggcttcagg tgggagcgcg 3121 tgatgaactt cgaggacggc ggcattattc acgttaacca ggactcctcc ctgcaggacg 3181 gcgtgttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa 3241 tgcagaagaa gaccatgggc tgggaggcct ccgaggagcg gatgtacccc gaggacggcg 3301 ccctgaagag cgagatcaag aagaggctga agctgaagga cggcggccac tacgccgccg 3361 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacatcgtcg 3421 acatcaagtt ggacatcgtg tcccacaacg aggactacac catcgtggaa cagtacgaac 3481 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaaggga gaagaagagt 3541 ttgatatgcc tcagcctcca catggccatg tcttacatcg tcacatgaga acaatccggg 3601 aagtacgcac acttgtcact cgtgtcatta cagatgtgta ttatgtggat ggaacagaag 3661 tagaaagaaa agtaactgag gagactgaag agccaattgt agagtgtcag gagtgtgaaa 3721 ctgaagtttc cccttcacag actgggggct cctcaggtga cctgggggat atcagctcct 3781 tctcctccaa ggcatccagc ttacaccgca catcaagtgg gacaagtctc tcagctatgc 3841 acagcagtgg aagctcaggg aaaggagccg gaccactcag agggaaaacc agcgggacag 3901 aacccgcaga ttttgcctta cccagctccc gaggaggccc aggaaaactg agtcctagaa 3961 aaggggtcag tcagacaggg acgccagtgt gtgaggagga tggtgatgca ggccttggca 4021 tcagacaggg agggaaggct ccagtcacgc ctcgtgggcg tgggcgaagg ggccgcccac 4081 cttctcggac cactggaacc agagaaacag ctgtgcctgg ccccttgggc atagaggaca 4141 tttcacctaa cttgtcacca gatgataaat ccttcagccg tgtcgtgccc cgagtgccag 4201 actccaccag acgaacagat gtgggtgctg gtgctttgcg tcgtagtgac tctccagaaa 4261 ttcctttcca ggctgctgct ggcccttctg atggcttaga tgcctcctct ccaggaaata 4321 gctttgtagg gctccgtgtt gtagccaagt ggtcatccaa tggctacttt tactctggga 4381 aaatcacacg agatgtcgga gctgggaagt ataaattgct ctttgatgat gggtacgaat 4441 gtgatgtgtt gggcaaagac attctgttat gtgaccccat cccgctggac actgaagtga 4501 cggccctctc ggaggatgag tatttcagtg caggagtggt gaaaggacat aggaaggagt 4561 ctggggaact gtactacagc attgaaaaag aaggccaaag aaagtggtat aagcgaatgg 4621 ctgtcatcct gtccttggag caaggaaaca gactgagaga gcagtatggg cttggcccct 4681 atgaagcagt aacacctctt acaaaggcag cagatatcag cttagacaat ttggtggaag 4741 ggaagcggaa acggcgcagt aacgtcagct ccccagccac ccctactgcc tccagtagca 4801 gcagcacaac ccctacccga aagatcacag aaagtcctcg tgcctccatg ggagttctct 4861 caggcaaaag aaaacttatc acttctgaag aggaacggtc ccctgccaag cgaggtcgca 4921 agtctgccac agtaaaacct ggtgcagtag gggcaggaga gtttgtgagc ccctgtgaga 4981 gtggagacaa caccgtgaat gatctagata attctaccgg gtaggggagg cgcttttccc 5041 aaggcagtct ggagcatgcg ctttagcagc cccgctgggc acttggcgct acacaagtgg 5101 cctctggcct cgcacacatt ccacatccac cggtaggcgc caaccggctc cgttctttgg 5161 tggccccttc gcgccacctt ctactcctcc cctagtcagg aagttccccc ccgccccgca 5221 gctcgcgtcg tgcaggacgt gacaaatgga agtagcacgt ctcactagtc tcgtgcagat 5281 ggacagcacc gctgagcaat ggaagcgggt aggcctttgg ggcagcggcc aatagcagct 5341 ttgctccttc gctttctggg ctcagaggct gggaaggggt gggtccgggg gcgggctcag 5401 gggcgggctc aggggcgggg cgggcgcccg aaggtcctcc ggaggcccgg cattctgcac 5461 gcttcaaaag cgcacgtctg ccgcgctgtt ctcctcttcc tcatctccgg gcctttcgac 5521 ctgcagccca agcttaccat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac 5581 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc 5641 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc 5701 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg 5761 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc 5821 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg 5881 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac 5941 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc 6001 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc 6061 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc 6121 cgcaagcccg gtgcctgacc gcgtctggaa caatcaacct ctggattaca aaatttgtga 6181 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt 6241 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa 6301 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt 6361 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct 6421 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg 6481 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc 6541 ggggaagctg acgtcctttc catggctgct cgcctgtgtt gccacctgga ttctgcgcgg 6601 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct 6661 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc 6721 cctttgggcc gcctccccgc ctggaattaa ttctgcagtc gagacctaga aaaacatgga 6781 gcaatcacaa gtagcaatac agcagctacc aatgctgatt gtgcctggct agaagcacaa 6841 gaggaggagg aggtgggttt tccagtcaca cctcaggtac ctttaagacc aatgacttac 6901 aaggcagctg tagatcttag ccacttttta aaagaaaaga ggggactgga agggctaatt 6961 cactcccaac gaagacaaga tatccttgat ctgtggatct accacacaca aggctacttc 7021 cctgattagc agaactacac accagggcca ggggtcagat atccactgac ctttggatgg 7081 tgctacaagc tagtaccagt tgagccagat aaggtagaag aggccaataa aggagagaac 7141 accagcttgt tacaccctgt gagcctgcat gggatggatg acccggagag agaagtgtta 7201 gagtggaggt ttgacagccg cctagcattt catcacgtgg cccgagagct gcatccggag 7261 tacttcaaga actgctgata tcgagcttgc tacaagggac tttccgctgg ggactttcca 7321 gggaggcgtg gcctgggcgg gactggggag tggcgagccc tcagatcctg catataagca 7381 gctgcttttt gcctgtactg ggtctctctg gttagaccag atctgagcct gggagctctc 7441 tggctaacta gggaacccac tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt 7501 agtgtgtgcc cgtctgttgt gtgactctgg taactagaga tccctcagac ccttttagtc 7561 agtgtggaaa atctctagca gtagtagttc atgtcatctt attattcagt atttataact 7621 tgcaaagaaa tgaatatcag agagtgagag gccttgacat tgctagcgtt ttaccgtcga 7681 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc 7741 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct 7801 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa 7861 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta 7921 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc 7981 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg 8041 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt 8101 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa 8161 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct 8221 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc 8281 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg 8341 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct 8401 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag 8461 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga 8521 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga 8581 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg 8641 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag 8701 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag 8761 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat 8821 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct 8881 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac 8941 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa 9001 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg 9061 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt 9121 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca 9181 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt 9241 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct 9301 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg 9361 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg 9421 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg 9481 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa 9541 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt 9601 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt 9661 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt 9721 gaatactcat actcttcctt ggtgatgctg ccaacttact gatttagtgt atgatggtgt 9781 ttttgaggtg ctccagtggc ttctgtttct atcagctgtc cctcctgttc agctactgac 9841 ggggtggtgc gtaacggcaa aagcaccgcc ggacatcagc gctatctctg ctctcactgc 9901 cgtaaaacat ggcaactgca gttcacttac accgcttctc aacccggtac gcaccagaaa 9961 atcattgata tggccatgaa tggcgttgga tgccgggcaa cagcccgcat tatgggcgtt 10021 ggcctcaaca cgattttacg tcacttaaaa aactcaggcc gcagtcggta acctcgcgca 10081 tacagccggg cagtgacgtc atcgtctgcg cggaaatgga cgaacagtgg ggctatgtcg 10141 gggctaaatc gcgccagcgc tggctgtttt acgcgtatga cagtctccgg aagacggttg 10201 ttgcgcacgt attcggtgaa cgcactatgg cgacgctggg gcgtcttatg agcctgctgt 10261 caccctttga cgtggtgata tggatgacgg atggctggcc gctgtatgaa tcccgcctga 10321 agggaaagct gcacgtaatc agcaagcgat atacgcagcg aattgagcgg cataacctga 10381 atctgaggca gcacctggca cggctgggac ggaagtcgct gtcgttctca aaatcggtgg 10441 agctgcatga caaagtcatc gggcattatc tgaacataaa acactatcaa taagttggag 10501 tcattaccct cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga 10561 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc 10621 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat 10681 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg 10741 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg 10801 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc 10861 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa 10921 agaaattgta tttgttaaat atgtactaca aacttagtag t //