质粒名称: | R4pGWB601_UBQ10p-miniTurbo-YFP-NLS |
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出品公司: | 爱迪基因 |
目录编号: | 127370 |
存储实验室: | Dominique Bergmann |
载体骨架基本信息 |
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载体名称: | R4pGWB601 |
载体出品公司: | -- |
载体抗性: | 大观霉素 |
空载体大小: | -- |
完整质粒大小: | 12680 bp |
载体修饰: | -- |
载体类型: | 植物表达 |
筛选标记: | Basta |
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: | miniTurbo (BirA mutant) |
物种来源: | BirA from E. coli, codon optimized for human and mutated |
插入大小: | 771 bp |
基因突变: | aa1-63 deleted; Q65P, I87V, R118S, E140K, Q141R, S150G, L151P, V160A, T192A, K194I, M209V, I305V |
GenBank编号 | -- |
启动子 | Arabidopsis UBQ10 promoter |
标签/融合蛋白: |
V5(插入N端子)
GS链接器(插入时为C终端)
YFP(插入件上的C端子)
NLS(插入的C端子)
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克隆方法: | Gateway 克隆 |
5' 克隆位点: | -- |
3' 克隆位点: | -- |
5' 测序引物 | M13 fw |
3' 测序引物 | M13 rv |
文献引用方法 |
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材料和方法部分: | R4pGWB601_UBQ10p-miniTurbo-YFP-NLS was a gift from Dominique Bergmann (Addgene plasmid # 127370 ; http://n2t.net/addgene:127370 ; RRID:Addgene_127370) |
参考文献部分: | Proximity labeling of protein complexes and cell type-specific organellar proteomes in Arabidopsis enabled by TurboID. Mair A, Xu SL, Branon TC, Ting AY, Bergmann DC. Elife. 2019 Sep 19;8. pii: 47864. doi: 10.7554/eLife.47864. 10.7554/eLife.47864 PubMed 31535972 |
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载体基本信息
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载体简介
载体序列
LOCUS R4pGWB601_UBQ10p-miniTurbo-YFP-NLS 12680 bp ds-DNA circular SYN 07-JUN-2019 DEFINITION Binary vector for expressing nuclear miniTurbo-YFP under the UBQ10 promoter in plants. ACCESSION . VERSION . KEYWORDS R4pGWB601_UBQ10p-miniTurbo-YFP-NLS SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 12680) TITLE Plasmids for proximity labeling with TurboID and miniTurbo JOURNAL Unpublished REFERENCE 2 (bases 1 to 12680) AUTHORS . TITLE Direct Submission JOURNAL Exported Jun 7, 2019 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..12680 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind 188..210 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 202..219 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 203..219 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" protein_bind 244..264 /gene="mutant version of attB" /label=attB4 /bound_moiety="BP Clonase(TM)" /note="core recombination site for the Gateway(R) BP reaction" protein_bind 2283..2307 /gene="mutant version of attB" /label=attB1 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 2321..2362 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" CDS 3167..3883 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3212..3233) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3473..3492) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3820..3841 /label=EGFP-C /note="EGFP, forward primer" CDS 3893..3913 /codon_start=1 /product="nuclear localization signal of SV40 (simian virus 40) large T antigen" /label=SV40 NLS /translation="PKKKRKV" protein_bind complement(3933..3957) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" terminator 3989..4241 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" primer_bind complement(4274..4290) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4274..4290) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4287..4309) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4298..4314 /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(4322..4352) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4367..4388 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." terminator 4424..4676 /label=NOS terminator /note="nopaline synthase terminator and poly(A) signal" CDS complement(4836..5387) /codon_start=1 /gene="Streptomyces hygroscopicus bar" /product="phosphinothricin acetyltransferase" /label=BlpR /note="confers resistance to bialophos or phosphinothricin" /translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF WQLDFSLPVPPRPVLPVTEI" promoter complement(5439..5618) /label=NOS promoter /note="nopaline synthase promoter" misc_feature 6124..6148 /label=LB T-DNA repeat /note="left border repeat from nopaline C58 T-DNA" CDS 6669..7460 /codon_start=1 /gene="aadA" /product="aminoglycoside adenylyltransferase (Murphy, 1985)" /label=SmR /note="confers resistance to spectinomycin and streptomycin" /translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK" rep_origin 7706..8294 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 8195..8214 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 8448..8465 /label=L4440 /note="L4440 vector, forward primer" misc_feature 8480..8620 /label=bom /note="basis of mobility region from pBR322" primer_bind complement(8546..8565) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 8706..8728 /label=pGEX 3' /note="pGEX vectors, reverse primer" rep_origin 8964..9158 /label=pVS1 oriV /note="origin of replication for the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" CDS complement(9224..10297) /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" CDS complement(10726..11355) /codon_start=1 /product="stability protein from the Pseudomonas plasmid pVS1 (Heeb et al., 2000)" /label=pVS1 StaA /translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI" misc_feature 12656..12680 /label=RB T-DNA repeat /note="right border repeat from nopaline C58 T-DNA" ORIGIN 1 aacactgata gtttaaactg aaggcgggaa acgacaatct gatccaagct caagctgctc 61 tagcattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg 121 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca 181 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagcttgt ggatccccca 241 tcacaacttt gtatagaaaa gttggctccg aattcgccct tagtctagct caacagagct 301 tttaacccaa attggtacaa tagaatacaa ctttagatca taattctcaa aagaaagaga 361 ttccttagct attctatctg ccactccatt tccttctcgg cttgtatgca caagcataaa 421 atcctcaaac ttgctaagta gatactttat gtcttggata attggattga gacttgacaa 481 gcataacttt catgtaacca aagacacaag ttgctgagaa tccacctcaa aaatgatctt 541 cctataattg aatcgggata atgacagcac agcccatcta agagcctcca cttctacttc 601 cagcacgctt cttactttta ccacagctct tgcacctaac cataacacct tccctgtatg 661 atcgcgaagc acccacccta agccacattt taatccttct gttggccatg ccccatcaaa 721 gttgcactta acccaagatt gtggtggagc ttcccatgtt tctcgtctgt cccgacggtg 781 ttgtggttgg tgctttcctt acattctgag cctctttcct tctaatccac tcatctgcat 841 cttcttgtgt ccttactaat acctcattgg ttccaaattc cctcccttta agcaccagct 901 cgtttctgtt cttccacagc ctcccaagta tccaagggac taaagcctcc acattcttca 961 gatcaggata ttcttgttta agatgttgaa ctctatggag gtttgtatga actgatgatc 1021 taggaccgga taagttccct tcttcatagc gaacttattc aaagaatgtt ttgtgtatca 1081 ttcttgttac attgttatta atgaaaaaat attattggtc attggactga acacgagtgt 1141 taaatatgga ccaggcccca aataagatcc attgatatat gaattaaata acaagaataa 1201 atcgagtcac caaaccactt gcctttttta acgagacttg ttcaccaact tgatacaaaa 1261 gtcattatcc tatgcaaatc aataatcata caaaaatatc caataacact aaaaaattaa 1321 aagaaatgga taatttcaca atatgttata cgataaagaa gttacttttc caagaaattc 1381 actgatttta taagcccact tgcattagat aaatggcaaa aaaaaacaaa aaggaaaaga 1441 aataaagcac gaagaattct agaaaatacg aaatacgctt caatgcagtg ggacccacgg 1501 ttcaattatt gccaattttc agctccaccg tatatttaaa aaataaaacg ataatgctaa 1561 aaaaatataa atcgtaacga tcgttaaatc tcaacggctg gatcttatga cgaccgttag 1621 aaattgtggt tgtcgacgag tcagtaataa acggcgtcaa agtggttgca gccggcacac 1681 acgagtcgtg tttatcaact caaagcacaa atacttttcc tcaacctaaa aataaggcaa 1741 ttagccaaaa acaactttgc gtgtaaacaa cgctcaatac acgtgtcatt ttattattag 1801 ctattgcttc accgccttag ctttctcgtg acctagtcgt cctcgtcttt tcttcttctt 1861 cttctataaa acaataccca aagagctctt cttcttcaca attcagattt caatttctca 1921 aaatcttaaa aactttctct caattctctc taccgtgatc aaggtaaatt tctgtgttcc 1981 ttattctctc aaaatcttcg attttgtttt cgttcgatcc caatttcgta tatgttcttt 2041 ggtttagatt ctgttaatct tagatcgaag acgattttct gggtttgatc gttagatatc 2101 atcttaattc tcgattaggg tttcatagat atcatccgat ttgttcaaat aatttgagtt 2161 ttgtcgaata attactcttc gatttgtgat ttctatctag atctggtgtt agtttctagt 2221 ttgtgcgatc gaatttgtcg attaatctga gtttttctga ttaacagaag ggcgaattcg 2281 acccaagttt gtacaaaaaa gcaggctccg cggccgcatg ggcaagccca tccccaaccc 2341 cctgctgggc ctggacagca ccgctagcat cccgctgctg aacgctaaac agattctggg 2401 acagctggac ggcgggagcg tggcagtcct gcctgtggtc gactccacca atcagtacct 2461 gctggatcga atcggcgagc tgaagagtgg ggatgcttgc attgcagaat atcagcaggc 2521 agggagagga agcagaggga ggaaatggtt ctctcctttt ggagctaacc tgtacctgag 2581 tatgttttgg cgcctgaagc ggggaccagc agcaatcggc ctgggcccgg tcatcggaat 2641 tgtcatggca gaagcgctgc gaaagctggg agcagacaag gtgcgagtca aatggcccaa 2701 tgacctgtat ctgcaggata gaaagctggc aggcatcctg gtggagctgg ccggaataac 2761 aggcgatgct gcacagatcg tcattggcgc cgggattaac gtggctatga ggcgcgtgga 2821 ggaaagcgtg gtcaatcagg gctggatcac actgcaggaa gcagggatta acctggacag 2881 gaatactctg gccgctatgc tgatccgaga gctgcgggca gccctggaac tgttcgagca 2941 ggaaggcctg gctccatatc tgtcacggtg ggagaagctg gataacttca tcaatagacc 3001 cgtgaagctg atcattgggg acaaagagat tttcgggatt agccggggga ttgataaaca 3061 gggagccctg ctgctggaac aggacggagt tatcaaaccc tggatgggcg gagaaatcag 3121 tctgcggtct gccgaaaagg gaggcggtgg aagcggcgga ggttccatgg tgagcaaggg 3181 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg 3241 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct 3301 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccacctt 3361 cggctacggc ctgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt 3421 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg 3481 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga 3541 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa 3601 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa 3661 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca 3721 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagctacca 3781 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt 3841 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aaggcttcgg atccaaaaaa 3901 gaagagaaag gtatagaagg gtgggcgcgc cgacccagct ttcttgtaca aagtggttga 3961 taacagcgct tagagctcga atttccccga tcgttcaaac atttggcaat aaagtttctt 4021 aagattgaat cctgttgccg gtcttgcgat gattatcata taatttctgt tgaattacgt 4081 taagcatgta ataattaaca tgtaatgcat gacgttattt atgagatggg tttttatgat 4141 tagagtcccg caattataca tttaatacgc gatagaaaac aaaatatagc gcgcaaacta 4201 ggataaatta tcgcgcgcgg tgtcatctat gttactagat cgggaattgg ttccggaacc 4261 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac 4321 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac 4381 tcacattagg ctgaattagg cgcgcctatt tctattaatt cccgatctag taacatagat 4441 gacaccgcgc gcgataattt atcctagttt gcgcgctata ttttgttttc tatcgcgtat 4501 taaatgtata attgcgggac tctaatcata aaaacccatc tcataaataa cgtcatgcat 4561 tacatgttaa ttattacatg cttaacgtaa ttcaacagaa attatatgat aatcatcgca 4621 agaccggcaa caggattcaa tcttaagaaa ctttattgcc aaatgtttga acgatcgggg 4681 aaattcgagc tggggacgtc cccagctcgg taccccctcc tgggcacggt cgtacgcgat 4741 ccgccgcggt tccacctcac cattgcgggt atgccagttg agttcgatga gttcccgtac 4801 cagctccaac tcgggcgatg ccacgcgcac tccgttcaga tctcggtgac gggcaggacc 4861 ggacggggcg gtaccggcag gctgaagtcc agctgccaga aacccacgtc atgccagttc 4921 ccgtgcttga agccggccgc ccgcagcatg ccgcgggggg catatccgag cgcctcgtgc 4981 atgcgcacgc tcgggtcgtt gggcagcccg atgacagcga ccacgctctt gaagccctgt 5041 gcctccaggg acttcagcag gtgggtgtag agcgtggagc ccagtcccgt ccgctggtgg 5101 cggggggaga cgtacacggt cgactcggcc gtccagtcgt aggcgttgcg tgccttccag 5161 gggcccgcgt aggcgatgcc ggcgacctcg ccgtccacct cggcgacgag ccagggatag 5221 cgctcccgca gacggacgag gtcgtccgtc cactcctgcg gttcctgcgg ctcggtacgg 5281 aagttgaccg tgcttgtctc gatgtagtgg ttgacgatgg tgcagaccgc cggcatgtcc 5341 gcctcggtgg cacggcggat gtcggccggg cgtcgttctg ggctcatatc ttattgcccc 5401 ccgggatcag atctggattg agagtgaata tgagactcta attggatacc gaggggaatt 5461 tatggaacgt cagtggagca tttttgacaa gaaatatttg ctagctgata gtgaccttag 5521 gcgacttttg aacgcgcaat aatggtttct gacgtatgtg cttagctcat taaactccag 5581 aaacccgcgg ctgagtggct ccttcaacgt tgcggttctg tcagttccaa acgtaaaacg 5641 gcttgtcccg cgtcatcggc gggggtcata acgtgactcc cttaattctc cgctcatgat 5701 cttgatcccc tgcgccatca gatccttggc ggcaagaaag ccatccagtt tactttgcag 5761 ggcttcccaa ccttaccaga gggcgcccca gctggcaatt ccggttcgct tgctgtccat 5821 aaaaccgccc agtctagcta tcgccatgta agcccactgc aagctacctg ctttctcttt 5881 gcgcttgcgt tttcccttgt ccagatagcc cagtagctga cattcatccg gggtcagcac 5941 cgtttctgcg gactggcttt ctacgtgttc cgcttccttt agcagccctt gcgccctgag 6001 tgcttgcggc agcgtgaagc tgatccgtcg agattaatag aaataaattc agcctaattc 6061 ggcgttaatt cagtacatta aaaacgtccg caatgtgtta ttaagttgtc taagcgtcaa 6121 tttgtttaca ccacaatata tcctgccacc agccagccaa cagctccccg accggcagct 6181 cggcacaaaa tcaccactcg atacaggcag cccatcagtc cgggacggcg tcagcgggag 6241 agccgttgta aggcggcaga ctttgctcat gttaccgatg ctattcggaa gaacggcaac 6301 taagctgccg ggtttgaaac acggatgatc tcgcggaggg tagcatgttg attgtaacga 6361 tgacagagcg ttgctgcctg tgatcaattc gggcacgaac ccagtggaca taagcctgtt 6421 cggttcgtaa gctgtaatgc aagtagcgta tgcgctcacg caactggtcc agaaccttga 6481 ccgaacgcag cggtggtaac ggcgcagtgg cggttttcat ggcttgttat gactgttttt 6541 ttggggtaca gtctatgcct cgggcatcca agcagcaagc gcgttacgcc gtgggtcgat 6601 gtttgatgtt atggagcagc aacgatgtta cgcagcaggg cagtcgccct aaaacaaagt 6661 taaacatcat gggggaagcg gtgatcgccg aagtatcgac tcaactatca gaggtagttg 6721 gcgtcatcga gcgccatctc gaaccgacgt tgctggccgt acatttgtac ggctccgcag 6781 tggatggcgg cctgaagcca cacagtgata ttgatttgct ggttacggtg accgtaaggc 6841 ttgatgaaac aacgcggcga gctttgatca acgacctttt ggaaacttcg gcttcccctg 6901 gagagagcga gattctccgc gctgtagaag tcaccattgt tgtgcacgac gacatcattc 6961 cgtggcgtta tccagctaag cgcgaactgc aatttggaga atggcagcgc aatgacattc 7021 ttgcaggtat cttcgagcca gccacgatcg acattgatct ggctatcttg ctgacaaaag 7081 caagagaaca tagcgttgcc ttggtaggtc cagcggcgga ggaactcttt gatccggttc 7141 ctgaacagga tctatttgag gcgctaaatg aaaccttaac gctatggaac tcgccgcccg 7201 actgggctgg cgatgagcga aatgtagtgc ttacgttgtc ccgcatttgg tacagcgcag 7261 taaccggcaa aatcgcgccg aaggatgtcg ctgccgactg ggcaatggag cgcctgccgg 7321 cccagtatca gcccgtcata cttgaagcta gacaggctta tcttggacaa gaagaagatc 7381 gcttggcctc gcgcgcagat cagttggaag aatttgtcca ctacgtgaaa ggcgagatca 7441 ccaaggtagt cggcaaataa tgtctagcta gaaattcgtt caagccgacg ccgcttcgcg 7501 gcgcggctta actcaagcgt tagatgcact aagcacataa ttgctcacag ccaaactatc 7561 aggtcaagtc tgcttttatt atttttaagc gtgcataata agccctacac aaattgggag 7621 atatatcatg catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc 7681 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct 7741 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa 7801 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag 7861 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc 7921 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg 7981 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca 8041 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat 8101 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg 8161 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc 8221 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc 8281 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc 8341 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg 8401 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga 8461 gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt 8521 cacaccgcat atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagt 8581 atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc cgccaacacc 8641 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac 8701 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca 8761 gggtgccttg atgtgggcgc cggcggtcga gtggcgacgg cgcggcttgt ccgcgccctg 8821 gtagattgcc tggccgtagg ccagccattt ttgagcggcc agcggccgcg ataggccgac 8881 gcgaagcggc ggggcgtagg gagcgcagcg accgaagggt aggcgctttt tgcagctctt 8941 cggctgtgcg ctggccagac agttatgcac aggccaggcg ggttttaaga gttttaataa 9001 gttttaaaga gttttaggcg gaaaaatcgc cttttttctc ttttatatca gtcacttaca 9061 tgtgtgaccg gttcccaatg tacggctttg ggttcccaat gtacgggttc cggttcccaa 9121 tgtacggctt tgggttccca atgtacgtgc tatccacagg aaagagacct tttcgacctt 9181 tttcccctgc tagggcaatt tgccctagca tctgctccgt acattaggaa ccggcggatg 9241 cttcgccctc gatcaggttg cggtagcgca tgactaggat cgggccagcc tgccccgcct 9301 cctccttcaa atcgtactcc ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac 9361 agaacttctt gaactctccg gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc 9421 tggcttctgc cttgcctgcg gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg 9481 gatcgatcaa aaagtaatcg gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga 9541 tctcgcggta catccaatca gctagctcga tctcgatgta ctccggccgc ccggtttcgc 9601 tctttacgat cttgtagcgg ctaatcaagg cttcaccctc ggataccgtc accaggcggc 9661 cgttcttggc cttcttcgta cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg 9721 tttctaccag gtcgtctttc tgctttccgc catcggctcg ccggcagaac ttgagtacgt 9781 ccgcaacgtg tggacggaac acgcggccgg gcttgtctcc cttcccttcc cggtatcggt 9841 tcatggattc ggttagatgg gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg 9901 ccatgccggc cggccctgcg gaaacctcta cgtgcccgtc tggaagctcg tagcggatca 9961 cctcgccagc tcgtcggtca cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc 10021 gactatcgcg ggtgcccacg tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc 10081 ccttgggcgg cttcctaatc gacggcgcac cggctgccgg cggttgccgg gattctttgc 10141 ggattcgatc agcggccgct tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt 10201 gccgctgggc ggcctgcgcg gccttcaact tctccaccag gtcatcaccc agcgccgcgc 10261 cgatttgtac cgggccggat ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt 10321 tccagtgcca ttgcagggcc ggcagacaac ccagccgctt acgcctggcc aaccgcccgt 10381 tcctccacac atggggcatt ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc 10441 gcctccttta gccgctaaaa ttcatctact catttattca tttgctcatt tactctggta 10501 gctgcgcgat gtattcagat agcagctcgg taatggtctt gccttggcgt accgcgtaca 10561 tcttcagctt ggtgtgatcc tccgccggca actgaaagtt gacccgcttc atggctggcg 10621 tgtctgccag gctggccaac gttgcagcct tgctgctgcg tgcgctcgga cggccggcac 10681 ttagcgtgtt tgtgcttttg ctcattttct ctttacctca ttaactcaaa tgagttttga 10741 tttaatttca gcggccagcg cctggacctc gcgggcagcg tcgccctcgg gttctgattc 10801 aagaacggtt gtgccggcgg cggcagtgcc tgggtagctc acgcgctgcg tgatacggga 10861 ctcaagaatg ggcagctcgt acccggccag cgcctcggca acctcaccgc cgatgcgcgt 10921 gcctttgatc gcccgcgaca cgacaaaggc cgcttgtagc cttccatccg tgacctcaat 10981 gcgctgctta accagctcca ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg 11041 caccggaatc agcacgaagt cggctgcctt gatcgcggac acagccaagt ccgccgcctg 11101 gggcgctccg tcgatcacta cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat 11161 cgtcgggcgg tcgatgccga caacggttag cggttgatct tcccgcacgg ccgcccaatc 11221 gcgggcactg ccctggggat cggaatcgac taacagaaca tcggccccgg cgagttgcag 11281 ggcgcgggct agatgggttg cgatggtcgt cttgcctgac ccgcctttct ggttaagtac 11341 agcgataacc ttcatgcgtt ccccttgcgt atttgtttat ttactcatcg catcatatac 11401 gcagcgaccg catgacgcaa gctgttttac tcaaatacac atcacctttt tagacggcgg 11461 cgctcggttt cttcagcggc caagctggcc ggccaggccg ccagcttggc atcagacaaa 11521 ccggccagga tttcatgcag ccgcacggtt gagacgtgcg cgggcggctc gaacacgtac 11581 ccggccgcga tcatctccgc ctcgatctct tcggtaatga aaaacggttc gtcctggccg 11641 tcctggtgcg gtttcatgct tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt 11701 cggcctcggt caatgcgtcc tcacggaagg caccgcgccg cctggcctcg gtgggcgtca 11761 cttcctcgct gcgctcaagt gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag 11821 ccgcctcttt cacggtgcgg ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg 11881 ccggggtgag ggtagggcgg gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc 11941 cgctccgggt gcggtcgatg attagggaac gctcgaactc ggcaatgccg gcgaacacgg 12001 tcaacaccat gcggccggcc ggcgtggtgg tgtcggccca cggctctgcc aggctacgca 12061 ggcccgcgcc ggcctcctgg atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg 12121 ccaggcggtc tagcctggtc actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc 12181 tggccagctc cgggcggtcg cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc 12241 agccggccgc gtgcagttcg gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc 12301 gggcatagcc cagcaggcca gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct 12361 agtcgcaagt attctacttt atgcgactaa aacacgcgac aagaaaacgc caggaaaagg 12421 gcagggcggc agcctgtcgc gtaacttagg acttgtgcga catgtcgttt tcagaagacg 12481 gctgcactga acgtcagaag ccgactgcac tatagcagcg gaggggttgg atcaaagtac 12541 tttgatcccg aggggaaccc tgtggttggc atgcacatac aaatggacga acggataaac 12601 cttttcacgc ccttttaaat atccgattat tctaataaac gctcttttct cttaggttta 12661 cccgccaata tatcctgtca //