质粒名称: | pLVX-puro_G-CEPIA1er |
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出品公司: | 爱迪基因 |
目录编号: | 164590 |
存储实验室: | Joseph Hyser |
载体骨架基本信息 |
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载体名称: | pLVX-puro |
载体出品公司: | Clontech |
载体抗性: | 氨苄青霉素 |
空载体大小: | 8037 bp |
完整质粒大小: | 9535 bp |
载体修饰: | -- |
载体类型: | 哺乳动物表达,慢病毒 |
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: | G-CEPIA1er |
物种来源: | Synthetic |
插入大小: | 1497 bp |
基因突变: | G-GECO1.1 E299D M304L D328N N345K F360W G369S E372D V395E D401E |
GenBank编号 | -- |
启动子 | CMV |
标签/融合蛋白: | Myc(插入件上的C端子) |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | XhoI(在克隆过程中被破坏) |
3' 克隆位点: | XbaI (未破坏) |
5' 测序引物 | CMV Forward |
3' 测序引物 | -- |
文献引用方法 |
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材料和方法部分: | pLVX-puro_G-CEPIA1er was a gift from Joseph Hyser (Addgene plasmid # 164590 ; http://n2t.net/addgene:164590 ; RRID:Addgene_164590) |
参考文献部分: | Rotavirus Calcium Dysregulation Manifests as Dynamic Calcium Signaling in the Cytoplasm and Endoplasmic Reticulum. Chang-Graham AL, Perry JL, Strtak AC, Ramachandran NK, Criglar JM, Philip AA, Patton JT, Estes MK, Hyser JM. Sci Rep. 2019 Jul 25;9(1):10822. doi: 10.1038/s41598-019-46856-8. 10.1038/s41598-019-46856-8 PubMed 31346185 |
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载体基本信息
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载体简介
载体序列
LOCUS pLVX-puro_G-CEPIA1er 9488 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Stable integration and expression of G-CEPIA1er in mammalian cells. ACCESSION . VERSION . KEYWORDS pLVX-puro_G-CEPIA1er SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 9488) AUTHORS Chang-Graham AL, Perry JL, Strtak AC, Ramachandran NK, Criglar JM, Philip AA, Patton JT, Estes MK, Hyser JM TITLE Rotavirus Calcium Dysregulation Manifests as Dynamic Calcium Signaling in the Cytoplasm and Endoplasmic Reticulum. JOURNAL Sci Rep. 2019 Jul 25;9(1):10822. doi: 10.1038/s41598-019-46856-8. PUBMED 31346185 REFERENCE 2 (bases 1 to 9488) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..9488 /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" regulatory 2821..2830 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" sig_peptide join(2827..2871,2954..2965) /product="modified signal sequence from the mouse immunoglobulin heavy chain variable region" /label=IgH signal sequence CDS 3002..3058 /codon_start=1 /product="calmodulin-binding peptide from avian smooth muscle myosin light chain kinase" /label=calmodulin-binding peptide /translation="RRKWNKTGHAVRAIGRLSS" primer_bind 3271..3292 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3659..3678) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" CDS 3794..4234 /codon_start=1 /product="calcium-binding protein" /label=calmodulin /translation="DQLTEEQIAEFKEAFSLFDKDGDGTITTKDLGTVLRSLGQNPTEA ELQDMINEVDADGNGTIDFPEFLTMMARKMKDTDSEEEIREAFRVWDKDGNGYISAADL RHVMTNLGEKLTDEEVDEMIREADIDGEGQVNYEEFVQMMTAK" CDS 4244..4273 /codon_start=1 /product="Myc (human c-Myc proto-oncogene) epitope tag" /label=Myc /translation="EQKLISEEDL" promoter 4323..4822 /label=PGK promoter /note="mouse phosphoglycerate kinase 1 promoter" primer_bind complement(4358..4381) /label=MSCV-rev /note="Murine stem cell virus, reverse primer" primer_bind 4755..4774 /label=mPGK-F /note="Mouse PGK promoter, forward primer" CDS 4843..5442 /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(4843..4862) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 5339..5359 /label=Puro-F /note="Puromycin resistance gene, forward primer" misc_feature 5456..6044 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5509..5529) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(5927..5938) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" LTR 6251..6884 /label=3' LTR /note="3' long terminal repeat (LTR) from HIV-1" primer_bind complement(7013..7029) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(7013..7029) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(7026..7048) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 7037..7053 /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(7061..7091) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 7106..7127 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(7244..7261) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7415..8003) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7495..7514) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8174..9034) /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 8797..8816 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(9035..9139) /gene="bla" /label=AmpR promoter polyA_signal 9187..9321 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(9224..9243) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 9278..9297 /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 taccggactc agatctcgag 2821 gccaccatgg gatggagctg tatcatcctc ttcttggtag caacagctac aggtaagggg 2881 ttaacagtag caggcttgag gtctggacat atatatgggt gacaatgaca tccactttgc 2941 ctttctctcc acaggcgcgc actcccaggt ccaactgcag ggatccatgg tcgactcttc 3001 acgtcgtaag tggaataaga caggtcacgc agtcagagct ataggtcggc tgagctcact 3061 cgagaacgtc tatatcaagg ccgacgagca gaagaacggc atcaaggcgt acttcaagat 3121 ccgccacaac atcgagggcg gcggcgtgca gctcgcctac cactaccagc agaacacccc 3181 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcgtgc agtccatact 3241 ttcgaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc 3301 cgggatcact ctcggcatgg acgagctgta caagggcggt accggaggga gcatggtgag 3361 caagggcgag gagctgttca ccggggtggt gcccatccag gtcgagctgg acggcgacgt 3421 aaacggccac aagttcagcg tgtccggcga gggtgagggc gatgccacct acggcaagct 3481 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac 3541 caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga 3601 cttcttcaag tccgccatgc ccgaaggcta catccaggag cgcaccatct tcttcaagga 3661 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg 3721 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga 3781 gtacaacacg cgtgaccaac tgactgaaga gcagatcgca gaatttaaag aggctttctc 3841 cctatttgac aaggacgggg atgggacgat aacaaccaag gatctgggga cggtgctgcg 3901 gtctctgggg cagaacccca cagaagcaga gctccaggac atgatcaatg aagtagatgc 3961 cgacggtaat ggcacaatcg acttccctga gttcctgaca atgatggcaa gaaaaatgaa 4021 agacacagac agtgaagaag aaattcgcga agcgttccgt gtgtgggata aggatggcaa 4081 tggctacatc tctgcagcag accttcgcca cgtgatgaca aaccttggag agaagttaac 4141 agatgaagag gttgatgaaa tgatcaggga agcagatatc gatggagaag gtcaggtaaa 4201 ctacgaagag tttgtacaaa tgatgacagc gaaggcggcc gcagaacaaa aactcatctc 4261 agaagaggat ctgaatgggg ccgcaagcga gaaggacgag ctgtagtcta gataattcta 4321 ccgggtaggg gaggcgcttt tcccaaggca gtctggagca tgcgctttag cagccccgct 4381 gggcacttgg cgctacacaa gtggcctctg gcctcgcaca cattccacat ccaccggtag 4441 gcgccaaccg gctccgttct ttggtggccc cttcgcgcca ccttctactc ctcccctagt 4501 caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg acgtgacaaa tggaagtagc 4561 acgtctcact agtctcgtgc agatggacag caccgctgag caatggaagc gggtaggcct 4621 ttggggcagc ggccaatagc agctttgctc cttcgctttc tgggctcaga ggctgggaag 4681 gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc ggggcgggcg cccgaaggtc 4741 ctccggaggc ccggcattct gcacgcttca aaagcgcacg tctgccgcgc tgttctcctc 4801 ttcctcatct ccgggccttt cgacctgcag cccaagctta ccatgaccga gtacaagccc 4861 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg 4921 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc 4981 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc 5041 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg 5101 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag 5161 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc 5221 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc 5281 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc 5341 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa 5401 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gaccgcgtct ggaacaatca 5461 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt 5521 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc 5581 tttcattttc tcctccttgt ataaatcctg gttgctgtct ctttatgagg agttgtggcc 5641 cgttgtcagg caacgtggcg tggtgtgcac tgtgtttgct gacgcaaccc ccactggttg 5701 gggcattgcc accacctgtc agctcctttc cgggactttc gctttccccc tccctattgc 5761 cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg acaggggctc ggctgttggg 5821 cactgacaat tccgtggtgt tgtcggggaa gctgacgtcc tttccatggc tgctcgcctg 5881 tgttgccacc tggattctgc gcgggacgtc cttctgctac gtcccttcgg ccctcaatcc 5941 agcggacctt ccttcccgcg gcctgctgcc ggctctgcgg cctcttccgc gtcttcgcct 6001 tcgccctcag acgagtcgga tctccctttg ggccgcctcc ccgcctggaa ttaattctgc 6061 agtcgagacc tagaaaaaca tggagcaatc acaagtagca atacagcagc taccaatgct 6121 gattgtgcct ggctagaagc acaagaggag gaggaggtgg gttttccagt cacacctcag 6181 gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt tttaaaagaa 6241 aagaggggac tggaagggct aattcactcc caacgaagac aagatatcct tgatctgtgg 6301 atctaccaca cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc 6361 agatatccac tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta 6421 gaagaggcca ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg 6481 gatgacccgg agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac 6541 gtggcccgag agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag 6601 ggactttccg ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga 6661 gccctcagat cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga 6721 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata 6781 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta 6841 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca 6901 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggccttg 6961 acattgctag cgttttaccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc 7021 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca 7081 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct 7141 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac 7201 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc 7261 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt 7321 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg 7381 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg 7441 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat 7501 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta 7561 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct 7621 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc 7681 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa 7741 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg 7801 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag 7861 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt 7921 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta 7981 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc 8041 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca 8101 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa 8161 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat 8221 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct 8281 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt 8341 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat 8401 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta 8461 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg 8521 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt 8581 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg 8641 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg 8701 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc 8761 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa 8821 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac 8881 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt 8941 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg 9001 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa 9061 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata 9121 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc gacggatcgg 9181 gagatcaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 9241 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 9301 gtatcttatc atgtctggat caactggata actcaagcta accaaaatca tcccaaactt 9361 cccaccccat accctattac cactgccaat tacctgtggt ttcatttact ctaaacctgt 9421 gattcctctg aattattttc attttaaaga aattgtattt gttaaatatg tactacaaac 9481 ttagtagt //