质粒名称: | pCas9-CAT |
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出品公司: | 爱迪基因 |
目录编号: | 80323 |
存储实验室: | Sebastian Lourido |
载体骨架基本信息 |
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载体名称: | pU6-Universal |
载体出品公司: | -- |
载体抗性: | 氨苄青霉素 |
空载体大小: | 8360 bp |
完整质粒大小: | 9301 bp |
载体修饰: | sgRNA区域已被氯霉素乙酰转移酶替换为TUB1 5’UTR和SAG1 3’UTR |
载体类型: | CRISPR |
筛选标记: | CAT |
高拷贝/低拷贝: | -- |
生长条件: | 37 ℃ |
插入基因信息 |
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插入基因名称: | Cas9 |
物种来源: | Streptococcus pyogenes |
插入大小: | 4101 bp |
基因突变: | -- |
GenBank编号 | -- |
启动子 | TgTUB1 |
标签/融合蛋白: |
3X标志(插入件上的N端子)
NLS(插入件上的N端子)
NLS(插接件上的C端子)
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克隆方法: | -- |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | CMV-F |
3' 测序引物 | BGH-rev |
文献引用方法 |
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材料和方法部分: | pCas9-CAT was a gift from Sebastian Lourido (Addgene plasmid # 80323 ; http://n2t.net/addgene:80323 ; RRID:Addgene_80323) |
参考文献部分: | A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P, Saeij JP, Carruthers VB, Niles JC, Lourido S. Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2. 10.1016/j.cell.2016.08.019 PubMed 27594426 |
买家导航
pCas9CAT载体基本信息
pCas9CAT载体图谱质粒图谱和多克隆位点信息
pCas9CAT载体简介
pCas9CAT载体序列
LOCUS pCas9-CAT 9301 bp ds-DNA circular SYN 06-AUG-2018 DEFINITION Encodes Cas9 and chloramphenicol acetyltransferase (CAT). ACCESSION . VERSION . KEYWORDS pCas9-CAT SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9301) AUTHORS Sidik SM, Huet D, Ganesan SM, Huynh MH, Wang T, Nasamu AS, Thiru P, Saeij JP, Carruthers VB, Niles JC, Lourido S TITLE A Genome-wide CRISPR Screen in Toxoplasma Identifies Essential Apicomplexan Genes. JOURNAL Cell. 2016 Sep 8;166(6):1423-1435.e12. doi: 10.1016/j.cell.2016.08.019. Epub 2016 Sep 2. PUBMED 27594426 REFERENCE 2 (bases 1 to 9301) AUTHORS . TITLE Direct Submission JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9301 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 509..574 /codon_start=1 /product="three tandem FLAG(R) epitope tags, followed by an enterokinase cleavage site" /label=3xFLAG /translation="DYKDHDGDYKDHDIDYKDDDDK" CDS 581..601 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" CDS 626..4726 /codon_start=1 /product="Cas9 (Csn1) endonuclease from the Streptococcus pyogenes Type II CRISPR/Cas system" /label=Cas9 /note="generates RNA-guided double strand breaks in DNA" /translation="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD ATLIHQSITGLYETRIDLSQLGGD" CDS 4727..4774 /codon_start=1 /product="bipartite nuclear localization signal from nucleoplasmin" /label=nucleoplasmin NLS /translation="KRPAATKKAGQAKKKK" primer_bind complement(4802..4819) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4808..5015 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 5024..5164 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 5024..5153 /label=AAV2 ITR rep_origin 5239..5694 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(5326..5345) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 5536..5557 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(5711..5730) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 5830..5852 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(5890..5908) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 5976..6080 /gene="bla" /label=AmpR promoter CDS 6081..6941 /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 complement(6299..6318) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 7112..7700 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 7601..7620 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 7773..7789 /label=pBluescriptKS /note="For pBluescript vector" primer_bind 7774..7790 /label=KS primer /note="common sequencing primer, one of multiple similar variants" CDS 8305..8967 /codon_start=1 /gene="cat" /product="chloramphenicol acetyltransferase" /label=CmR /note="confers resistance to chloramphenicol" /translation="MHEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFS SLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVAN MDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA" primer_bind complement(8374..8393) /label=CAT-R /note="Chloramphenicol resistance gene, reverse primer" primer_bind complement(9285..9301) /label=SK primer /note="common sequencing primer, one of multiple similar variants" primer_bind complement(9285..9301) /label=pBluescriptSK /note="For pBluescript vector" ORIGIN 1 ggtaccatgc atgtcccgcg ttcgtgaaat tctctgcatc agcggagtga tcaggaatca 61 tcgtctcagc gggatgacgt tgcggagcag gccggctcgc ggtgggcagt cagatgccga 121 aggcgtaact caggacggct tgcgctcatc gcagaacagg ggtggtgcct gcattgggtg 181 cggttggtga tcctggttgg accggtggag atgcgcgcgc acgaagggga tgtgtcagaa 241 acattttgtt tgttctctgt gaacttttag atgtgttaaa ggcggcgaat attancagag 301 agtcctcctt gttggattct ctcttgaatt tcgccctttc cttctctttg cgagtctcgt 361 agagaacaag cactcgttcg ccgtccctga cgacgcaacc cgcgcagaag acatccacca 421 aacggtgtta cacaatcacc ttgtgtgaag ttcttgcgga aaactactcg ttggcatttt 481 ttcttgaatt ccctttttcg acaccatgga ctataaggac cacgacggag actacaagga 541 tcatgatatt gattacaaag acgatgacga taagatggcc ccaaagaaga agcggaaggt 601 cggtatccac ggagtcccag cagccgacaa gaagtacagc atcggcctgg acatcggcac 661 caactctgtg ggctgggccg tgatcaccga cgagtacaag gtgcccagca agaaattcaa 721 ggtgctgggc aacaccgacc ggcacagcat caagaagaac ctgatcggag ccctgctgtt 781 cgacagcggc gaaacagccg aggccacccg gctgaagaga accgccagaa gaagatacac 841 cagacggaag aaccggatct gctatctgca agagatcttc agcaacgaga tggccaaggt 901 ggacgacagc ttcttccaca gactggaaga gtccttcctg gtggaagagg ataagaagca 961 cgagcggcac cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc 1021 caccatctac cacctgagaa agaaactggt ggacagcacc gacaaggccg acctgcggct 1081 gatctatctg gccctggccc acatgatcaa gttccggggc cacttcctga tcgagggcga 1141 cctgaacccc gacaacagcg acgtggacaa gctgttcatc cagctggtgc agacctacaa 1201 ccagctgttc gaggaaaacc ccatcaacgc cagcggcgtg gacgccaagg ccatcctgtc 1261 tgccagactg agcaagagca gacggctgga aaatctgatc gcccagctgc ccggcgagaa 1321 gaagaatggc ctgttcggaa acctgattgc cctgagcctg ggcctgaccc ccaacttcaa 1381 gagcaacttc gacctggccg aggatgccaa actgcagctg agcaaggaca cctacgacga 1441 cgacctggac aacctgctgg cccagatcgg cgaccagtac gccgacctgt ttctggccgc 1501 caagaacctg tccgacgcca tcctgctgag cgacatcctg agagtgaaca ccgagatcac 1561 caaggccccc ctgagcgcct ctatgatcaa gagatacgac gagcaccacc aggacctgac 1621 cctgctgaaa gctctcgtgc ggcagcagct gcctgagaag tacaaagaga ttttcttcga 1681 ccagagcaag aacggctacg ccggctacat tgacggcgga gccagccagg aagagttcta 1741 caagttcatc aagcccatcc tggaaaagat ggacggcacc gaggaactgc tcgtgaagct 1801 gaacagagag gacctgctgc ggaagcagcg gaccttcgac aacggcagca tcccccacca 1861 gatccacctg ggagagctgc acgccattct gcggcggcag gaagattttt acccattcct 1921 gaaggacaac cgggaaaaga tcgagaagat cctgaccttc cgcatcccct actacgtggg 1981 ccctctggcc aggggaaaca gcagattcgc ctggatgacc agaaagagcg aggaaaccat 2041 caccccctgg aacttcgagg aagtggtgga caagggcgct tccgcccaga gcttcatcga 2101 gcggatgacc aacttcgata agaacctgcc caacgagaag gtgctgccca agcacagcct 2161 gctgtacgag tacttcaccg tgtataacga gctgaccaaa gtgaaatacg tgaccgaggg 2221 aatgagaaag cccgccttcc tgagcggcga gcagaaaaag gccatcgtgg acctgctgtt 2281 caagaccaac cggaaagtga ccgtgaagca gctgaaagag gactacttca agaaaatcga 2341 gtgcttcgac tccgtggaaa tctccggcgt ggaagatcgg ttcaacgcct ccctgggcac 2401 ataccacgat ctgctgaaaa ttatcaagga caaggacttc ctggacaatg aggaaaacga 2461 ggacattctg gaagatatcg tgctgaccct gacactgttt gaggacagag agatgatcga 2521 ggaacggctg aaaacctatg cccacctgtt cgacgacaaa gtgatgaagc agctgaagcg 2581 gcggagatac accggctggg gcaggctgag ccggaagctg atcaacggca tccgggacaa 2641 gcagtccggc aagacaatcc tggatttcct gaagtccgac ggcttcgcca acagaaactt 2701 catgcagctg atccacgacg acagcctgac ctttaaagag gacatccaga aagcccaggt 2761 gtccggccag ggcgatagcc tgcacgagca cattgccaat ctggccggca gccccgccat 2821 taagaagggc atcctgcaga cagtgaaggt ggtggacgag ctcgtgaaag tgatgggccg 2881 gcacaagccc gagaacatcg tgatcgaaat ggccagagag aaccagacca cccagaaggg 2941 acagaagaac agccgcgaga gaatgaagcg gatcgaagag ggcatcaaag agctgggcag 3001 ccagatcctg aaagaacacc ccgtggaaaa cacccagctg cagaacgaga agctgtacct 3061 gtactacctg cagaatgggc gggatatgta cgtggaccag gaactggaca tcaaccggct 3121 gtccgactac gatgtggacc atatcgtgcc tcagagcttt ctgaaggacg actccatcga 3181 caacaaggtg ctgaccagaa gcgacaagaa ccggggcaag agcgacaacg tgccctccga 3241 agaggtcgtg aagaagatga agaactactg gcggcagctg ctgaacgcca agctgattac 3301 ccagagaaag ttcgacaatc tgaccaaggc cgagagaggc ggcctgagcg aactggataa 3361 ggccggcttc atcaagagac agctggtgga aacccggcag atcacaaagc acgtggcaca 3421 gatcctggac tcccggatga acactaagta cgacgagaat gacaagctga tccgggaagt 3481 gaaagtgatc accctgaagt ccaagctggt gtccgatttc cggaaggatt tccagtttta 3541 caaagtgcgc gagatcaaca actaccacca cgcccacgac gcctacctga acgccgtcgt 3601 gggaaccgcc ctgatcaaaa agtaccctaa gctggaaagc gagttcgtgt acggcgacta 3661 caaggtgtac gacgtgcgga agatgatcgc caagagcgag caggaaatcg gcaaggctac 3721 cgccaagtac ttcttctaca gcaacatcat gaactttttc aagaccgaga ttaccctggc 3781 caacggcgag atccggaagc ggcctctgat cgagacaaac ggcgaaaccg gggagatcgt 3841 gtgggataag ggccgggatt ttgccaccgt gcggaaagtg ctgagcatgc cccaagtgaa 3901 tatcgtgaaa aagaccgagg tgcagacagg cggcttcagc aaagagtcta tcctgcccaa 3961 gaggaacagc gataagctga tcgccagaaa gaaggactgg gaccctaaga agtacggcgg 4021 cttcgacagc cccaccgtgg cctattctgt gctggtggtg gccaaagtgg aaaagggcaa 4081 gtccaagaaa ctgaagagtg tgaaagagct gctggggatc accatcatgg aaagaagcag 4141 cttcgagaag aatcccatcg actttctgga agccaagggc tacaaagaag tgaaaaagga 4201 cctgatcatc aagctgccta agtactccct gttcgagctg gaaaacggcc ggaagagaat 4261 gctggcctct gccggcgaac tgcagaaggg aaacgaactg gccctgccct ccaaatatgt 4321 gaacttcctg tacctggcca gccactatga gaagctgaag ggctcccccg aggataatga 4381 gcagaaacag ctgtttgtgg aacagcacaa gcactacctg gacgagatca tcgagcagat 4441 cagcgagttc tccaagagag tgatcctggc cgacgctaat ctggacaaag tgctgtccgc 4501 ctacaacaag caccgggata agcccatcag agagcaggcc gagaatatca tccacctgtt 4561 taccctgacc aatctgggag cccctgccgc cttcaagtac tttgacacca ccatcgaccg 4621 gaagaggtac accagcacca aagaggtgct ggacgccacc ctgatccacc agagcatcac 4681 cggcctgtac gagacacgga tcgacctgtc tcagctggga ggcgacaaaa ggccggcggc 4741 cacgaaaaag gccggccagg caaaaaagaa aaagtaagaa ttcctagagc tcgctgatca 4801 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc 4861 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg 4921 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg 4981 gaggattggg aagagaatag caggcatgct ggggagcggc cgcaggaacc cctagtgatg 5041 gagttggcca ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc 5101 gcccgacgcc cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg 5161 caggggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata 5221 cgtcaaagca accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg 5281 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct 5341 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc 5401 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg 5461 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt 5521 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg 5581 gctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc 5641 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt 5701 gcactctcag tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa 5761 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg 5821 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga 5881 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt 5941 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt 6001 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat 6061 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt 6121 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg 6181 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga 6241 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc 6301 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac 6361 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg 6421 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca 6481 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg 6541 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg 6601 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg 6661 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag 6721 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg 6781 gagccggtga gcgtggaagc cgcggtatca ttgcagcact ggggccagat ggtaagccct 6841 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac 6901 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact 6961 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga 7021 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt 7081 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct 7141 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc 7201 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc 7261 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc 7321 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg 7381 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt 7441 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg 7501 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg 7561 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt 7621 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag 7681 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt 7741 gctggccttt tgctcacatg tcgggccccc cctcgaggtc gacggtatcg ataagcttga 7801 tggcgatgca tgtcccgcgt tcgtgaaatt ctctgcatca gcggagtgat caggaatcat 7861 cgtctcagcg ggatgacgtt gcggagcagg ccggctcgcg gtgggcagtc agatgccgaa 7921 ggcgtaactc aggacggctt gcgctcatcg cagaacaggg gtggtgcctg cattgggtgc 7981 ggttggtgat cctggttgga ccggtggaga tgcgcgcgca cgaaggggat gtgtcagaaa 8041 cattttgttt gttctctgtg aacttttaga tgtgttaaag gcggcgaata ttagcagaga 8101 gtcctccttg ttggattctc tcttgaattt cgccctttcc ttctctttgc gagtctcgta 8161 gagaacaagc actcgttcgc cgtccctgac gacgcaaccc gcgcagaaga catccaccaa 8221 acggtgttac acaatcacct tgtgtgaagt tcttgcggaa aactactcgt tggcattttt 8281 tcttgaattc cctttttcga caaaatgcat gagaaaaaaa tcactggata taccaccgtt 8341 gatatatccc aatggcatcg taaagaacat tttgaggcat ttcagtcagt tgctcaatgt 8401 acctataacc agaccgttca gctggatatt acggcctttt taaagaccgt aaagaaaaat 8461 aagcacaagt tttatccggc atttattcac attcttgccc gcctgatgaa tgctcatccg 8521 gaattccgta tggcaatgaa agacggtgag ctggtgatat gggatagtgt tcacccttgt 8581 tacaccgttt tccatgagca aactgaaacg ttttcatcgc tctggagtga ataccacgac 8641 gatttccggc agtttctaca catatattcg caagatgtgg cgtgttacgg tgaaaacctg 8701 gcctatttcc ctaaagggtt tattgagaat atgtttttcg tctcagccaa tccctgggtg 8761 agtttcacca gttttgattt aaacgtggcc aatatggaca acttcttcgc ccccgttttc 8821 accatgggca aatattatac gcaaggcgac aaggtgctga tgccgctggc gattcaggtt 8881 catcatgccg tttgtgatgg cttccatgtc ggcagaatgc ttaatgaatt acaacagtac 8941 tgcgatgagt ggcagggcgg ggcttaatta atcaccgttg tgctcacttc tcaaatcgac 9001 aaaggaaaca cacttcgtgc agcatgtgcc ccattataaa gaaactgagt tgttccgttg 9061 tggcttgcag gtgtcacatc cacaaaaacc ggccgactct aaataggagt gtttcgcagc 9121 aagcagcgaa agtttatgac tgggtccgaa tctctgaacg gatgtgtggc ggacctggct 9181 gatgttgatc gccgtcgaca cacgcgccac atgggtcaat acacaagaca gctatcagtt 9241 gttttagtcg aaccggttaa cacaattctt gcccccccga gggggatcca ctagttctag 9301 a //