质粒名称: | pMK381 (AAVS1 CMV-OsTIR1F74G) |
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出品公司: | 爱迪基因 |
目录编号: | 140536 |
存储实验室: | Masato Kanemaki |
载体骨架基本信息 |
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载体名称: | pMK359 |
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: | 嘌呤霉素 |
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: | CMV-OsTIR1(F74G) |
物种来源: | H. sapiens (human) |
插入大小: | -- |
基因突变: | -- |
GenBank编号 | -- |
启动子 | -- |
标签/融合蛋白: | -- |
克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | -- |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pMK381 (AAVS1 CMV-OsTIR1F74G) was a gift from Masato Kanemaki (Addgene plasmid # 140536 ; http://n2t.net/addgene:140536 ; RRID:Addgene_140536) |
参考文献部分: | The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT.. Nat Commun (2020) 11(1):5701 10.1038/s41467-020-19532-z |
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载体基本信息
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载体简介
载体序列
LOCUS pMK381 (AAVS1 CMV-OsTIR1F74G) 10098 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAVS1 CMV-OsTIR1(F74G). ACCESSION . VERSION . KEYWORDS pMK381 (AAVS1 CMV-OsTIR1F74G) SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10098) AUTHORS Yesbolatova A, Saito Y, Kitamoto N, Makino-Itou H, Ajima R, Nakano R, Nakaoka H, Fukui K, Gamo K, Tominari Y, Takeuchi H, Saga Y, Hayashi KI, Kanemaki MT. TITLE The auxin-inducible degron 2 technology provides sharp degradation control in yeast, mammalian cells, and mice. JOURNAL Nat Commun (2020) 11(1):5701 REFERENCE 2 (bases 1 to 10098) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10098 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 107..128 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 143..173 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 181..197 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." primer_bind 186..208 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 205..221 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 205..221 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" promoter 242..260 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" misc_feature 295..1098 /label=HA-L /note="left homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" enhancer 1153..1456 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 1457..1660 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 1611..1631 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" polyA_signal 3599..3720 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(3636..3655) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 3690..3709 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind complement(3747..3780) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." promoter 3841..4340 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(3876..3899) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4273..4292 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4380..4979 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APHNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(4380..4399) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" protein_bind complement(5536..5569) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 5600..6436 /label=HA-R /note="right homology arm from the adeno-associated virus integration site (AAVS1) within intron 1 of the human PPP1R12C gene" primer_bind complement(6470..6489) /label=T7 /note="T7 promoter, forward primer" promoter complement(6471..6489) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(6496..6513) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(6496..6512) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6505..6527) /label=M13/pUC Forward /note="In lacZ gene" CDS 6650..6952 /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="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI" primer_bind 6831..6850 /label=ccdB-fwd /note="ccdB gene, forward primer" CDS 7301..8095 /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="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(7355..7374) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 7965..7984 /label=Neo-F /note="Neomycin resistance gene, forward primer" CDS complement(8345..9205) /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 8968..8987 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 9329..9917 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 9818..9837 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 10071..10088 /label=L4440 /note="L4440 vector, forward primer" ORIGIN 1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc 61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc 121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa 181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctca 241 gaattaaccc tcactaaagg gactagtcct gcaggtttaa acgaattcgc cctttgcttt 301 ctctgaccag cattctctcc cctgggcctg tgccgctttc tgtctgcagc ttgtggcctg 361 ggtcacctct acggctggcc cagatccttc cctgccgcct ccttcaggtt ccgtcttcct 421 ccactccctc ttccccttgc tctctgctgt gttgctgccc aaggatgctc tttccggagc 481 acttccttct cggcgctgca ccacgtgatg tcctctgagc ggatcctccc cgtgtctggg 541 tcctctccgg gcatctctcc tccctcaccc aaccccatgc cgtcttcact cgctgggttc 601 ccttttcctt ctccttctgg ggcctgtgcc atctctcgtt tcttaggatg gccttctccg 661 acggatgtct cccttgcgtc ccgcctcccc ttcttgtagg cctgcatcat caccgttttt 721 ctggacaacc ccaaagtacc ccgtctccct ggctttagcc acctctccat cctcttgctt 781 tctttgcctg gacaccccgt tctcctgtgg attcgggtca cctctcactc ctttcatttg 841 ggcagctccc ctacccccct tacctctcta gtctgtgcta gctcttccag ccccctgtca 901 tggcatcttc caggggtccg agagctcagc tagtcttctt cctccaaccc gggcccctat 961 gtccacttca ggacagcatg tttgctgcct ccagggatcc tgtgtccccg agctgggacc 1021 accttatatt cccagggccg gttaatgtgg ctctggttct gggtactttt atctgtcccc 1081 tccaccccac agtggggcaa gctagtaatc aattacgggg tcattagttc atagcccata 1141 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga 1201 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt 1261 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt 1321 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca 1381 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt 1441 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt 1501 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca 1561 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg 1621 cggtaggcgt gtacggtggg aggtctatat aagcagagct ggtttagtga accgtcagat 1681 ccgctagcgc tgccaccatg acatactttc ctgaagaggt cgtcgaacac atttttagct 1741 tcctgcctgc acagagagat agaaacacag tgagcctggt ctgcaaagtg tggtacgaga 1801 tcgaacgcct gagccggaga ggagtgttcg tcggcaactg ctatgctgtg agagcaggca 1861 gggtcgccgc taggtttcca aatgtgcgcg cactgaccgt caaggggaaa ccccacggcg 1921 ccgactttaa cctggtgccc cctgattggg gaggatacgc cggcccttgg atcgaggcag 1981 ccgctcgcgg ctgtcatgga ctggaggaac tgcgcatgaa gcgaatggtg gtctctgacg 2041 aaagtctgga gctgctggct cggagcttcc ctaggtttcg cgcactggtg ctgatttctt 2101 gcgaaggctt cagcaccgat ggactggcag ccgtggcctc ccactgtaag ctgctgcggg 2161 agctggacct ccaggagaat gaagtggagg atagaggccc cagatggctg tcttgcttcc 2221 cagactcatg taccagcctg gtgtccctga actttgcctg catcaaaggc gaagtgaatg 2281 ctgggtccct ggagcggctg gtctcaagaa gccccaacct gaggtctctg cggctgaacc 2341 ggagcgtgag cgtggacact ctggctaaga ttctgctgag aacccctaac ctggaggatc 2401 tgggaaccgg caatctgaca gacgatttcc agacagaatc ctactttaaa ctgacttctg 2461 ccctggagaa gtgtaaaatg ctgaggagtc tgtcaggatt ctgggatgct tcacccgtgt 2521 gcctgagctt tatctaccct ctgtgtgcac agctgacagg cctgaacctg agctatgcac 2581 caaccctgga cgccagtgat ctgacaaaga tgatctcacg ctgcgtgaaa ctccagcgac 2641 tgtgggtgct ggactgtatt tccgataagg ggctccaggt ggtcgccagc tcctgcaagg 2701 acctccagga gctgagagtg ttcccatctg atttttacgt ggccggatat agtgctgtca 2761 ctgaggaagg cctggtggca gtctcactgg gatgcccaaa gctgaacagc ctgctgtatt 2821 tctgtcatca gatgactaat gctgcactgg tgaccgtcgc caagaactgc cctaatttca 2881 cccgatttcg gctgtgtatt ctggaaccag gcaaacccga cgtggtcaca tcccagccac 2941 tggatgaagg gtttggagct atcgtgagag agtgcaaggg actccagagg ctgagcattt 3001 ccggcctgct gacagacaaa gtgttcatgt acatcggcaa gtatgctaag cagctggaga 3061 tgctgagcat tgcatttgcc ggagactccg ataagggcat gatgcacgtg atgaacgggt 3121 gtaagaatct gcgaaaactg gaaatccggg acagcccttt cggggatgcc gctctgctgg 3181 gaaactttgc cagatacgag acaatgagga gcctgtggat gtctagttgc aatgtgactc 3241 tgaagggctg tcaggtcctg gctagtaaaa tgcctatgct gaacgtggaa gtcattaatg 3301 agcgggacgg gtctaacgaa atggaggaaa atcatggcga cctgccaaag gtggagaaac 3361 tgtatgtgta tcggaccacc gcaggggcaa gagatgatgc tcccaacttt gtgaagattc 3421 tgtgaggatc catcggatcc cgggcccgtc gacggtaccg cgggcccggg atccaccggg 3481 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa 3541 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa 3601 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa 3661 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta 3721 agcgatcata ttcaataacc cttaatataa cttcgtataa tgtatgctat acgaagttat 3781 taggtctgaa gaggagttta cgtccagcca agcttaggat ctcgacctcg aaattctacc 3841 gggtagggga ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg 3901 gcacttggcg ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc 3961 gccaaccgac tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca 4021 ggaagttccc ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac 4081 gtctcactag tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt 4141 ggggcagcgg ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg 4201 gtgggtccgg gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct 4261 ccggaggccc ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt 4321 cctcatctcc gggcctttcg acctgcatcc atctagatct cgagcagctg aagcttacca 4381 tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac 4441 gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc gatccggacc 4501 gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc gggctcgaca 4561 tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc acgccggaga 4621 gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag ttgagcggtt 4681 cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg cccaaggagc 4741 ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag ggtctgggca 4801 gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg 4861 agacctccgc gccccacaac ctccccttct acgagcggct cggcttcacc gtcaccgccg 4921 acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgac 4981 gcccgcccca cgacccgcag cgcccgaccg aaaggagcgc acgaccccat gcatcgatga 5041 tatcagatcc ccgggatgca gaaattgatg atctattaaa caataaagat gtccactaaa 5101 atggaagttt ttcctgtcat actttgttaa gaagggtgag aacagagtac ctacattttg 5161 aatggaagga ttggagctac gggggtgggg gtggggtggg attagataaa tgcctgctct 5221 ttactgaagg ctctttacta ttgctttatg ataatgtttc atagttggat atcataattt 5281 aaacaagcaa aaccaaatta agggccagct cattcctccc actcatgatc tatagatcta 5341 tagatctctc gtgggatcat tgtttttctc ttgattccca ctttgtggtt ctaagtactg 5401 tggtttccaa atgtgtcagt ttcatagcct gaagaacgag atcagcagcc tctgttccac 5461 atacacttca ttctcagtat tgttttgcca agttctaatt ccatcagaag ctggtcgaga 5521 tccggaaccc ttaatataac ttcgtataat gtatgctata cgaagttatt aggtccctcg 5581 aagaggttca ctaagcttta ctagggacag gattggtgac agaaaagccc catccttagg 5641 cctcctcctt cctagtctcc tgatattggg tctaaccccc acctcctgtt aggcagattc 5701 cttatctggt gacacacccc catttcctgg agccatctct ctccttgcca gaacctctaa 5761 ggtttgctta cgatggagcc agagaggatc ctgggaggga gagcttggca gggggtggga 5821 gggaaggggg ggatgcgtga cctgcccggt tctcagtggc caccctgcgc taccctctcc 5881 cagaacctga gctgctctga cgcggctgtc tggtgcgttt cactgatcct ggtgctgcag 5941 cttccttaca cttcccaaga ggagaagcag tttggaaaaa caaaatcaga ataagttggt 6001 cctgagttct aactttggct cttcaccttt ctagtcccca atttatattg ttcctccgtg 6061 cgtcagtttt acctgtgaga taaggccagt agccagcccc gtcctggcag ggctgtggtg 6121 aggagggggg tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc taggtgttca 6181 ccaggtcgtg gccgcctcta ctccctttct ctttctccat ccttctttcc ttaaagagtc 6241 cccagtgcta tctgggacat attcctccgc ccagagcagg gtcccgcttc cctaaggccc 6301 tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc aggcttccct 6361 gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc cctacagggg 6421 ttcctggctc tgctctaagg gcgaattcgc ggccgctaaa ttcaattcgc cctatagtga 6481 gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt 6541 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga 6601 ggcccgcacc gatcgccctt cccaacagtt gcgcagccta tacgtacggc agtttaaggt 6661 ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga gtgatattat 6721 tgacacgccg gggcgacgga tggtgatccc cctggccagt gcacgtctgc tgtcagataa 6781 agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc gcatgatgac 6841 caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg atctcagcca 6901 ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat aaatgtcagg 6961 catgagatta tcaaaaagga tcttcaccta gatccttttc acgtagaaag ccagtccgca 7021 gaaacggtgc tgaccccgga tgaatgtcag ctactgggct atctggacaa gggaaaacgc 7081 aagcgcaaag agaaagcagg tagcttgcag tgggcttaca tggcgatagc tagactgggc 7141 ggttttatgg acagcaagcg aaccggaatt gccagctggg gcgccctctg gtaaggttgg 7201 gaagccctgc aaagtaaact ggatggcttt cttgccgcca aggatctgat ggcgcagggg 7261 atcaagctct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt 7321 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca 7381 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct 7441 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct 7501 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc 7561 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct 7621 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga 7681 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg 7741 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc 7801 agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac 7861 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat 7921 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga 7981 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc 8041 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgaattat 8101 taacgcttac aatttcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca 8161 ccgcatcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct 8221 aaatacattc aaatatgtat ccgctcatga gattatcaaa aaggatcttc acctagatcc 8281 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg 8341 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat 8401 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg 8461 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa 8521 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca 8581 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc 8641 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt 8701 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa 8761 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat 8821 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct 8881 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga 8941 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag 9001 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga 9061 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca 9121 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg 9181 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc 9241 agggttattg tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag 9301 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct 9361 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac 9421 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc 9481 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg 9541 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt 9601 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt 9661 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc 9721 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca 9781 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata 9841 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg 9901 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct 9961 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta 10021 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag 10081 tgagcgagga agcggaag //