质粒名称: | pGFPGUSPlus |
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出品公司: | Addgene |
目录编号: | 64401 |
存储实验室: | Claudia Vickers |
载体骨架基本信息 |
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载体名称: | pCAMBIA1305.1 |
载体出品公司: | CAMBIA |
载体抗性: | 卡那霉素 |
空载体大小: | -- |
完整质粒大小: | -- |
载体修饰: | -- |
载体类型: | 植物表达;植物转化的二元载体 |
筛选标记: | 潮霉素 |
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: | GFP |
物种来源: | -- |
插入大小: | -- |
基因突变: | S65T |
GenBank编号 | ABQ22954.1 |
启动子 | 35S |
标签/融合蛋白: | -- |
克隆方法: | 限制性内切酶 |
5' 克隆位点: | HindIII(未破坏) |
3' 克隆位点: | EcoRI(未破坏) |
5' 测序引物 | M13-F20 |
3' 测序引物 | M13-R |
文献引用方法 |
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材料和方法部分: | pGFPGUSPlus was a gift from Claudia Vickers (Addgene plasmid # 64401 ; http://n2t.net/addgene:64401 ; RRID:Addgene_64401) |
参考文献部分: | pGFPGUSPlus, a new binary vector for gene expression studies and optimising transformation systems in plants. Vickers CE, Schenk PM, Li D, Mullineaux PM, Gresshoff PM. Biotechnol Lett. 2007 Nov;29(11):1793-6. Epub 2007 Aug 9. 10.1007/s10529-007-9467-6 PubMed 17687623 |
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载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS pGFPGUSPlus 13702 bp ds-DNA circular SYN 07-DEC-2018 DEFINITION binary plant vector expressing GFP and GUS, both driven by a 35S promoter. ACCESSION . VERSION . KEYWORDS pGFPGUSPlus SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 13702) AUTHORS Vickers CE, Schenk PM, Li D, Mullineaux PM, Gresshoff PM TITLE pGFPGUSPlus, a new binary vector for gene expression studies and optimising transformation systems in plants. JOURNAL Biotechnol Lett. 2007 Nov;29(11):1793-6. Epub 2007 Aug 9. PUBMED 17687623 REFERENCE 2 (bases 1 to 13702) AUTHORS . TITLE Direct Submission JOURNAL Exported Dec 7, 2018 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..13702 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(85..102) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(85..101) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(94..116) /label=M13/pUC Forward /note="In lacZ gene" promoter 478..823 /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" CDS join(840..854,1045..2862) /codon_start=1 /product="beta-glucuronidase" /label=GUSPlus(TM) /note="codon-optimized Staphylococcus gusA gene with a catalase intron to ensure expression in plants but not bacteria" /translation="MVDLRNRRTSLYPINTETRGVFDLNGVWNFKLDYGKGLEEKWYES KLTDTISMAVPSSYNDIGVTKEIRNHIGYVWYEREFTVPAYLKDQRIVLRFGSATHKAI VYVNGELVVEHKGGFLPFEAEINNSLRDGMNRVTVAVDNILDDSTLPVGLYSERHEEGL GKVIRNKPNFDFFNYAGLHRPVKIYTTPFTYVEDISVVTDFNGPTGTVTYTVDFQGKAE TVKVSVVDEEGKVVASTEGLSGNVEIPNVILWEPLNTYLYQIKVELVNDGLTIDVYEEP FGVRTVEVNDGKFLINNKPFYFKGFGKHEDTPINGRGFNEASNVMDFNILKWIGANSFR TAHYPYSEELMRLADREGLVVIDETPAVGVHLNFMATTGLGEGSERVSTWEKIRTFEHH QDVLRELVSRDKNHPSVVMWSIANEAATEEEGAYEYFKPLVELTKELDPQKRPVTIVLF VMATPETDKVAELIDVIALNRYNGWYFDGGDLEAAKVHLRQEFHAWNKRCPGKPIMITE YGADTVAGFHDIDPVMFTEEYQVEYYQANHVVFDEFENFVGEQAWNFADFATSQGVMRV QGNKKGVFTRDRKPKLAAHVFRERWTNIPDFGYKN" intron 855..1044 /label=cat1 intron /note="castor bean catalase intron, modified" CDS 2869..2886 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" terminator 2918..3170 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" misc_feature 3192..3216 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" CDS 4517..5146 /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" CDS 5575..6648 /codon_start=1 /product="replication protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 RepA /translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI LVMRYRNLIEGEASAGS" rep_origin 6714..6908 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" primer_bind complement(7144..7166) /label=pGEX 3' /note="pGEX vectors, reverse primer" misc_feature 7252..7392 /label=bom /note="basis of mobility region from pBR322" primer_bind 7307..7326 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind complement(7407..7424) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7578..8166) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7658..7677) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8253..9047) /codon_start=1 /gene="aphA-3" /product="aminoglycoside phosphotransferase" /label=KanR /note="confers resistance to kanamycin" /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF" misc_feature 9472..9496 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" polyA_signal 9574..9748 /label=CaMV poly(A) signal /note="cauliflower mosaic virus polyadenylation signal" CDS complement(9789..10814) /codon_start=1 /gene="aph(4)-Ia" /product="aminoglycoside phosphotransferase from E. coli" /label=HygR /note="confers resistance to hygromycin" /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK K" promoter complement(10881..11557) /label=CaMV 35S promoter (enhanced) /note="cauliflower mosaic virus 35S promoter with a duplicated enhancer region" protein_bind 11748..11769 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 11784..11814 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 11822..11838 /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 11827..11849 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 11846..11862 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 11846..11862 /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" terminator 11879..12131 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(12177..12896) /codon_start=1 /product="enhanced GFP" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITHGMDELYK" primer_bind complement(12222..12243) /label=EGFP-C /note="EGFP, forward primer" primer_bind 12571..12590 /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 12830..12851 /label=EGFP-N /note="EGFP, reverse primer" promoter complement(12927..13272) /label=CaMV 35S promoter /note="strong constitutive promoter from cauliflower mosaic virus" ORIGIN 1 aagcctctct aaccatctgt gggttagcat tctttctgaa attgaaaagg ctaatctggg 61 gacctgcagg catgcaagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa 121 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa 181 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg 241 ctagagcagc ttgagcttgg atcagattgt cgtttcccgc cttcagttta gcttcatgga 301 gtcaaagatt caaatagagg acctaacaga actcgccgta aagactggcg aacagttcat 361 acagagtctc ttacgactca atgacaagaa gaaaatcttc gtcaacatgg tggagcacga 421 cacacttgtc tactccaaaa atatcaaaga tacagtctca gaagaccaaa gggcaattga 481 gacttttcaa caaagggtaa tatccggaaa cctcctcgga ttccattgcc cagctatctg 541 tcactttatt gtgaagatag tggaaaagga aggtggctcc tacaaatgcc atcattgcga 601 taaaggaaag gccatcgttg aagatgcctc tgccgacagt ggtcccaaag atggaccccc 661 acccacgagg agcatcgtgg aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga 721 ttgatgtgat atctccactg acgtaaggga tgacgcacaa tcccactatc cttcgcaaga 781 cccttcctct atataaggaa gttcatttca tttggagaga acacggggga ctcttgacca 841 tggtagatct gagggtaaat ttctagtttt tctccttcat tttcttggtt aggacccttt 901 tctcttttta tttttttgag ctttgatctt tctttaaact gatctatttt ttaattgatt 961 ggttatggtg taaatattac atagctttaa ctgataatct gattacttta tttcgtgtgt 1021 ctatgatgat gatgatagtt acagaaccga cgaactagtc tgtacccgat caacaccgag 1081 acccgtggcg tcttcgacct caatggcgtc tggaacttca agctggacta cgggaaagga 1141 ctggaagaga agtggtacga aagcaagctg accgacacta ttagtatggc cgtcccaagc 1201 agttacaatg acattggcgt gaccaaggaa atccgcaacc atatcggata tgtctggtac 1261 gaacgtgagt tcacggtgcc ggcctatctg aaggatcagc gtatcgtgct ccgcttcggc 1321 tctgcaactc acaaagcaat tgtctatgtc aatggtgagc tggtcgtgga gcacaagggc 1381 ggattcctgc cattcgaagc ggaaatcaac aactcgctgc gtgatggcat gaatcgcgtc 1441 accgtcgccg tggacaacat cctcgacgat agcaccctcc cggtggggct gtacagcgag 1501 cgccacgaag agggcctcgg aaaagtcatt cgtaacaagc cgaacttcga cttcttcaac 1561 tatgcaggcc tgcaccgtcc ggtgaaaatc tacacgaccc cgtttacgta cgtcgaggac 1621 atctcggttg tgaccgactt caatggccca accgggactg tgacctatac ggtggacttt 1681 caaggcaaag ccgagaccgt gaaagtgtcg gtcgtggatg aggaaggcaa agtggtcgca 1741 agcaccgagg gcctgagcgg taacgtggag attccgaatg tcatcctctg ggaaccactg 1801 aacacgtatc tctaccagat caaagtggaa ctggtgaacg acggactgac catcgatgtc 1861 tatgaagagc cgttcggcgt gcggaccgtg gaagtcaacg acggcaagtt cctcatcaac 1921 aacaaaccgt tctacttcaa gggctttggc aaacatgagg acactcctat caacggccgt 1981 ggctttaacg aagcgagcaa tgtgatggat ttcaatatcc tcaaatggat cggcgccaac 2041 agcttccgga ccgcacacta tccgtactct gaagagttga tgcgtcttgc ggatcgcgag 2101 ggtctggtcg tgatcgacga gactccggca gttggcgtgc acctcaactt catggccacc 2161 acgggactcg gcgaaggcag cgagcgcgtc agtacctggg agaagattcg gacgtttgag 2221 caccatcaag acgttctccg tgaactggtg tctcgtgaca agaaccatcc aagcgtcgtg 2281 atgtggagca tcgccaacga ggcggcgact gaggaagagg gcgcgtacga gtacttcaag 2341 ccgttggtgg agctgaccaa ggaactcgac ccacagaagc gtccggtcac gatcgtgctg 2401 tttgtgatgg ctaccccgga gacggacaaa gtcgccgaac tgattgacgt catcgcgctc 2461 aatcgctata acggatggta cttcgatggc ggtgatctcg aagcggccaa agtccatctc 2521 cgccaggaat ttcacgcgtg gaacaagcgt tgcccaggaa agccgatcat gatcactgag 2581 tacggcgcag acaccgttgc gggctttcac gacattgatc cagtgatgtt caccgaggaa 2641 tatcaagtcg agtactacca ggcgaaccac gtcgtgttcg atgagtttga gaacttcgtg 2701 ggtgagcaag cgtggaactt cgcggacttc gcgacctctc agggcgtgat gcgcgtccaa 2761 ggaaacaaga agggcgtgtt cactcgtgac cgcaagccga agctcgccgc gcacgtcttt 2821 cgcgagcgct ggaccaacat tccagatttc ggctacaaga acgctagcca tcaccatcac 2881 catcacgtgt gaattggtga ccagctcgaa tttccccgat cgttcaaaca tttggcaata 2941 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt 3001 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt 3061 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg 3121 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattaaa 3181 ctatcagtgt ttgacaggat atattggcgg gtaaacctaa gagaaaagag cgtttattag 3241 aataatcgga tatttaaaag ggcgtgaaaa ggtttatccg ttcgtccatt tgtatgtgca 3301 tgccaaccac agggttcccc tcgggatcaa agtactttga tccaacccct ccgctgctat 3361 agtgcagtcg gcttctgacg ttcagtgcag ccgtcttctg aaaacgacat gtcgcacaag 3421 tcctaagtta cgcgacaggc tgccgccctg cccttttcct ggcgttttct tgtcgcgtgt 3481 tttagtcgca taaagtagaa tacttgcgac tagaaccgga gacattacgc catgaacaag 3541 agcgccgccg ctggcctgct gggctatgcc cgcgtcagca ccgacgacca ggacttgacc 3601 aaccaacggg ccgaactgca cgcggccggc tgcaccaagc tgttttccga gaagatcacc 3661 ggcaccaggc gcgaccgccc ggagctggcc aggatgcttg accacctacg ccctggcgac 3721 gttgtgacag tgaccaggct agaccgcctg gcccgcagca cccgcgacct actggacatt 3781 gccgagcgca tccaggaggc cggcgcgggc ctgcgtagcc tggcagagcc gtgggccgac 3841 accaccacgc cggccggccg catggtgttg accgtgttcg ccggcattgc cgagttcgag 3901 cgttccctaa tcatcgaccg cacccggagc gggcgcgagg ccgccaaggc ccgaggcgtg 3961 aagtttggcc cccgccctac cctcaccccg gcacagatcg cgcacgcccg cgagctgatc 4021 gaccaggaag gccgcaccgt gaaagaggcg gctgcactgc ttggcgtgca tcgctcgacc 4081 ctgtaccgcg cacttgagcg cagcgaggaa gtgacgccca ccgaggccag gcggcgcggt 4141 gccttccgtg aggacgcatt gaccgaggcc gacgccctgg cggccgccga gaatgaacgc 4201 caagaggaac aagcatgaaa ccgcaccagg acggccagga cgaaccgttt ttcattaccg 4261 aagagatcga ggcggagatg atcgcggccg ggtacgtgtt cgagccgccc gcgcacgtct 4321 caaccgtgcg gctgcatgaa atcctggccg gtttgtctga tgccaagctg gcggcctggc 4381 cggccagctt ggccgctgaa gaaaccgagc gccgccgtct aaaaaggtga tgtgtatttg 4441 agtaaaacag cttgcgtcat gcggtcgctg cgtatatgat gcgatgagta aataaacaaa 4501 tacgcaaggg gaacgcatga aggttatcgc tgtacttaac cagaaaggcg ggtcaggcaa 4561 gacgaccatc gcaacccatc tagcccgcgc cctgcaactc gccggggccg atgttctgtt 4621 agtcgattcc gatccccagg gcagtgcccg cgattgggcg gccgtgcggg aagatcaacc 4681 gctaaccgtt gtcggcatcg accgcccgac gattgaccgc gacgtgaagg ccatcggccg 4741 gcgcgacttc gtagtgatcg acggagcgcc ccaggcggcg gacttggctg tgtccgcgat 4801 caaggcagcc gacttcgtgc tgattccggt gcagccaagc ccttacgaca tatgggccac 4861 cgccgacctg gtggagctgg ttaagcagcg cattgaggtc acggatggaa ggctacaagc 4921 ggcctttgtc gtgtcgcggg cgatcaaagg cacgcgcatc ggcggtgagg ttgccgaggc 4981 gctggccggg tacgagctgc ccattcttga gtcccgtatc acgcagcgcg tgagctaccc 5041 aggcactgcc gccgccggca caaccgttct tgaatcagaa cccgagggcg acgctgcccg 5101 cgaggtccag gcgctggccg ctgaaattaa atcaaaactc atttgagtta atgaggtaaa 5161 gagaaaatga gcaaaagcac aaacacgcta agtgccggcc gtccgagcgc acgcagcagc 5221 aaggctgcaa cgttggccag cctggcagac acgccagcca tgaagcgggt caactttcag 5281 ttgccggcgg aggatcacac caagctgaag atgtacgcgg tacgccaagg caagaccatt 5341 accgagctgc tatctgaata catcgcgcag ctaccagagt aaatgagcaa atgaataaat 5401 gagtagatga attttagcgg ctaaaggagg cggcatggaa aatcaagaac aaccaggcac 5461 cgacgccgtg gaatgcccca tgtgtggagg aacgggcggt tggccaggcg taagcggctg 5521 ggttgcctgc cggccctgca atggcactgg aacccccaag cccgaggaat cggcgtgagc 5581 ggtcgcaaac catccggccc ggtacaaatc ggcgcggcgc tgggtgatga cctggtggag 5641 aagttgaagg ccgcgcaggc cgcccagcgg caacgcatcg aggcagaagc acgccccggt 5701 gaatcgtggc aagcggccgc tgatcgaatc cgcaaagaat cccggcaacc gccggcagcc 5761 ggtgcgccgt cgattaggaa gccgcccaag ggcgacgagc aaccagattt tttcgttccg 5821 atgctctatg acgtgggcac ccgcgatagt cgcagcatca tggacgtggc cgttttccgt 5881 ctgtcgaagc gtgaccgacg agctggcgag gtgatccgct acgagcttcc agacgggcac 5941 gtagaggttt ccgcagggcc ggccggcatg gccagtgtgt gggattacga cctggtactg 6001 atggcggttt cccatctaac cgaatccatg aaccgatacc gggaagggaa gggagacaag 6061 cccggccgcg tgttccgtcc acacgttgcg gacgtactca agttctgccg gcgagccgat 6121 ggcggaaagc agaaagacga cctggtagaa acctgcattc ggttaaacac cacgcacgtt 6181 gccatgcagc gtacgaagaa ggccaagaac ggccgcctgg tgacggtatc cgagggtgaa 6241 gccttgatta gccgctacaa gatcgtaaag agcgaaaccg ggcggccgga gtacatcgag 6301 atcgagctag ctgattggat gtaccgcgag atcacagaag gcaagaaccc ggacgtgctg 6361 acggttcacc ccgattactt tttgatcgat cccggcatcg gccgttttct ctaccgcctg 6421 gcacgccgcg ccgcaggcaa ggcagaagcc agatggttgt tcaagacgat ctacgaacgc 6481 agtggcagcg ccggagagtt caagaagttc tgtttcaccg tgcgcaagct gatcgggtca 6541 aatgacctgc cggagtacga tttgaaggag gaggcggggc aggctggccc gatcctagtc 6601 atgcgctacc gcaacctgat cgagggcgaa gcatccgccg gttcctaatg tacggagcag 6661 atgctagggc aaattgccct agcaggggaa aaaggtcgaa aaggtctctt tcctgtggat 6721 agcacgtaca ttgggaaccc aaagccgtac attgggaacc ggaacccgta cattgggaac 6781 ccaaagccgt acattgggaa ccggtcacac atgtaagtga ctgatataaa agagaaaaaa 6841 ggcgattttt ccgcctaaaa ctctttaaaa cttattaaaa ctcttaaaac ccgcctggcc 6901 tgtgcataac tgtctggcca gcgcacagcc gaagagctgc aaaaagcgcc tacccttcgg 6961 tcgctgcgct ccctacgccc cgccgcttcg cgtcggccta tcgcggccgc tggccgctca 7021 aaaatggctg gcctacggcc aggcaatcta ccagggcgcg gacaagccgc gccgtcgcca 7081 ctcgaccgcc ggcgcccaca tcaaggcacc ctgcctcgcg cgtttcggtg atgacggtga 7141 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg 7201 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gcgcagccat 7261 gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc atcagagcag 7321 attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa 7381 taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg 7441 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg 7501 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag 7561 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga 7621 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct 7681 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc 7741 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg 7801 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc 7861 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca 7921 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag 7981 ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct 8041 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc 8101 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga 8161 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca 8221 cgttaaggga ttttggtcat gcattctagg tactaaaaca attcatccag taaaatataa 8281 tattttattt tctcccaatc aggcttgatc cccagtaagt caaaaaatag ctcgacatac 8341 tgttcttccc cgatatcctc cctgatcgac cggacgcaga aggcaatgtc ataccacttg 8401 tccgccctgc cgcttctccc aagatcaata aagccactta ctttgccatc tttcacaaag 8461 atgttgctgt ctcccaggtc gccgtgggaa aagacaagtt cctcttcggg cttttccgtc 8521 tttaaaaaat catacagctc gcgcggatct ttaaatggag tgtcttcttc ccagttttcg 8581 caatccacat cggccagatc gttattcagt aagtaatcca attcggctaa gcggctgtct 8641 aagctattcg tatagggaca atccgatatg tcgatggagt gaaagagcct gatgcactcc 8701 gcatacagct cgataatctt ttcagggctt tgttcatctt catactcttc cgagcaaagg 8761 acgccatcgg cctcactcat gagcagattg ctccagccat catgccgttc aaagtgcagg 8821 acctttggaa caggcagctt tccttccagc catagcatca tgtccttttc ccgttccaca 8881 tcataggtgg tccctttata ccggctgtcc gtcattttta aatataggtt ttcattttct 8941 cccaccagct tatatacctt agcaggagac attccttccg tatcttttac gcagcggtat 9001 ttttcgatca gttttttcaa ttccggtgat attctcattt tagccattta ttatttcctt 9061 cctcttttct acagtattta aagatacccc aagaagctaa ttataacaag acgaactcca 9121 attcactgtt ccttgcattc taaaacctta aataccagaa aacagctttt tcaaagttgt 9181 tttcaaagtt ggcgtataac atagtatcga cggagccgat tttgaaaccg cggtgatcac 9241 aggcagcaac gctctgtcat cgttacaatc aacatgctac cctccgcgag atcatccgtg 9301 tttcaaaccc ggcagcttag ttgccgttct tccgaatagc atcggtaaca tgagcaaagt 9361 ctgccgcctt acaacggctc tcccgctgac gccgtcccgg actgatgggc tgcctgtatc 9421 gagtggtgat tttgtgccga gctgccggtc ggggagctgt tggctggctg gtggcaggat 9481 atattgtggt gtaaacaaat tgacgcttag acaacttaat aacacattgc ggacgttttt 9541 aatgtactga attaacgccg aattaattcg ggggatctgg attttagtac tggattttgg 9601 ttttaggaat tagaaatttt attgatagaa gtattttaca aatacaaata catactaagg 9661 gtttcttata tgctcaacac atgagcgaaa ccctatagga accctaattc ccttatctgg 9721 gaactactca cacattatta tggagaaact cgagcttgtc gatcgacaga tcccggtcgg 9781 catctactct atttctttgc cctcggacga gtgctggggc gtcggtttcc actatcggcg 9841 agtacttcta cacagccatc ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc 9901 ccgacagtcc cggctccgga tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca 9961 tcatcgaaat tgccgtcaac caagctctga tagagttggt caagaccaat gcggagcata 10021 tacgcccgga gtcgtggcga tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc 10081 tgctgctcca tacaagccaa ccacggcctc cagaagaaga tgttggcgac ctcgtattgg 10141 gaatccccga acatcgcctc gctccagtca atgaccgctg ttatgcggcc attgtccgtc 10201 aggacattgt tggagccgaa atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg 10261 gcccaaagca tcagctcatc gagagcctgc gcgacggacg cactgacggt gtcgtccatc 10321 acagtttgcc agtgatacac atggggatca gcaatcgcgc atatgaaatc acgccatgta 10381 gtgtattgac cgattccttg cggtccgaat gggccgaacc cgctcgtctg gctaagatcg 10441 gccgcagcga tcgcatccat agcctccgcg accggttgta gaacagcggg cagttcggtt 10501 tcaggcaggt cttgcaacgt gacaccctgt gaacggcggg agatgcaata ggtcaggctc 10561 tcgctaaact ccccaatgtc aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc 10621 cgataaacat aacgatcttt gtagaaacca tcggcgcagc tatttacccg caggacatat 10681 ccacgccctc ctacatcgaa gctgaaagca cgagattctt cgccctccga gagctgcatc 10741 aggtcggaga cgctgtcgaa cttttcgatc agaaacttct cgacagacgt cgcggtgagt 10801 tcaggctttt tcatatctca ttgccccccg gatctgcgaa agctcgagag agatagattt 10861 gtagagagag actggtgatt tcagcgtgtc ctctccaaat gaaatgaact tccttatata 10921 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata 10981 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 11041 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc 11101 tttcctttat cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg 11161 atgaagtgac agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt 11221 tgaaaagtct caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag 11281 acgagagtgt cgtgctccac catgttcaca tcaatccact tgctttgaag acgtggttgg 11341 aacgtcttct ttttccacga tgctcctcgt gggtgggggt ccatctttgg gaccactgtc 11401 ggcagaggca tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc 11461 accttccttt tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag 11521 gaggtttccc gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac 11581 tgtatctttg atattcttgg agtagacgag agtgtcgtgc tccaccatgt tggcaagctg 11641 ctctagccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg 11701 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag 11761 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga 11821 attgtgagcg gataacaatt tcacacagga aacagctatg acatgattac gaattcccga 11881 tctagtaaca tagatgacac cgcgcgcgat aatttatcct agtttgcgcg ctatattttg 11941 ttttctatcg cgtattaaat gtataattgc gggactctaa tcataaaaac ccatctcata 12001 aataacgtca tgcattacat gttaattatt acatgcttaa cgtaattcaa cagaaattat 12061 atgataatca tcgcaagacc ggcaacagga ttcaatctta agaaacttta ttgccaaatg 12121 tttgaacgat cggggaaatt cgagctcgga tcaattcctg cagccgggcg gccgctttac 12181 ttgtacagct cgtccatgcc gtgagtgatc ccggcggcgg tcacgaactc cagcaggacc 12241 atgtgatcgc gcttctcgtt ggggtctttg ctcagggcgg actgggtgct caggtagtgg 12301 ttgtcgggca gcagcacggg gccgtcgccg atgggggtgt tctgctggta gtggtcggcg 12361 agctgcacgc tgccgtcctc gatgttgtgg cggatcttga agttcacctt gatgccgttc 12421 ttctgcttgt cggccatgat atagacgttg tggctgttgt agttgtactc cagcttgtgc 12481 cccaggatgt tgccgtcctc cttgaagtcg atgcccttca gctcgatgcg gttcaccagg 12541 gtgtcgccct cgaacttcac ctcggcgcgg gtcttgtagt tgccgtcgtc cttgaagaag 12601 atggtgcgct cctggacgta gccttcgggc atggcggact tgaagaagtc gtgctgcttc 12661 atgtggtcgg ggtagcggct gaagcactgc acgccgtagg tgaaggtggt cacgagggtg 12721 ggccagggca cgggcagctt gccggtggtg cagatgaact tcagggtcag cttgccgtag 12781 gtggcatcgc cctcgccctc gccggacacg ctgaacttgt ggccgtttac gtcgccgtcc 12841 agctcgacca ggatgggcac caccccggtg aacagctcct cgcccttgct caccatggga 12901 tcgaattgat cctctagagt cccccgtgtt ctctccaaat gaaatgaact tccttatata 12961 gaggaagggt cttgcgaagg atagtgggat tgtgcgtcat cccttacgtc agtggagata 13021 tcacatcaat ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc 13081 ctcgtgggtg ggggtccatc tttgggacca ctgtcggcag aggcatcttc aacgatggcc 13141 tttcctttat cgcaatgatg gcatttgtag gagccacctt ccttttccac tatcttcaca 13201 ataaagtgac agatagctgg gcaatggaat ccgaggaggt ttccggatat taccctttgt 13261 tgaaaagtct caattgccct ttggtcttct gagactgtat ctttgatatt tttggagtag 13321 acaagtgtgt cgtgctccac catgttgacg aagattttct tcttgtcatt gagtcgtaag 13381 agactctgta tgaactgttc gccagtcttt acggcgagtt ctgttaggtc ctctatttga 13441 atctttgact ccatggcctt tgattcagtg ggaactacct ttttagagac tccaatctct 13501 attacttgcc ttggtttgtg aagcaagcct tgaatcgtcc atactggaat agtacttctg 13561 atcttgagaa atatatcttt ctctgtgttc ttgatgcagt tagtcctgaa tcttttgact 13621 gcatctttaa ccttcttggg aaggtatttg atttcctgga gattattgct cgggtagatc 13681 gtcttgatga gacctgctgc gt //