| 质粒名称: | pRGEB31 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 51295 |
| 存储实验室: | Yinong Yang |
|
载体骨架基本信息 |
|
| 载体名称: | pCAMBIA1300 |
| 载体出品公司: | CAMBIA |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 8958 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 1. Removed Bsa I sites. 2. Inserted the U3-gRNA-35S-Cas9 cassette from pRGE31. |
| 载体类型: | CRISPR;植物表达 |
| 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 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | 水稻snoRNA U3和双35S启动子 |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | M13R(-48) |
| 3' 测序引物 | M13F(-20) |
|
文献引用方法 |
|
| 材料和方法部分: | pRGEB31 was a gift from Yinong Yang (Addgene plasmid # 51295 ; http://n2t.net/addgene:51295 ; RRID:Addgene_51295) |
| 参考文献部分: | RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System. Xie K, Yang Y. Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013 Aug 17. 10.1093/mp/sst119 PubMed 23956122 |
买家导航
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 pRGEB31 15037 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Binary vector derived from pRGE31. Deliver sgRNA and Cas9 into
plant. by agrobacterium mediated transformation..
ACCESSION .
VERSION .
KEYWORDS pRGEB31
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15037)
AUTHORS Xie K, Yang Y
TITLE RNA-Guided Genome Editing in Plants Using a CRISPR-Cas System.
JOURNAL Mol Plant. 2013 Nov;6(6):1975-83. doi: 10.1093/mp/sst119. Epub 2013
Aug 17.
PUBMED 23956122
REFERENCE 2 (bases 1 to 15037)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..15037
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 6..386
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
misc_RNA 403..478
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 1044..1389
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
primer_bind 1332..1352
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
protein_bind 1411..1435
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 1480..1545
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1552..1572
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1597..5697
/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 5698..5745
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
protein_bind complement(5778..5802)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 5813..5836
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
terminator 5858..6110
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
primer_bind complement(6119..6135)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6136..6152)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6145..6167)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 6355..6379
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 7679..8308
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 8742..9809
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="GRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESWQA
AADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLSKR
DRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKPGR
VFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGEAL
ISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYRLA
RRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPILV
MRYRNLIEGEASAGS"
rep_origin 9875..10069
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(10305..10327)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 10413..10553
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 10468..10487
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(10568..10585)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10739..11327)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10819..10838)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(11414..12208)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 12633..12657
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 12735..12909
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(12949..13974)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(14042..14719)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
protein_bind 14910..14931
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 14946..14976
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 14984..15000
/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 14989..15011
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 15008..15024
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 15008..15024
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 agcttaagga atctttaaac atacgaacag atcacttaaa gttcttctga agcaacttaa
61 agttatcagg catgcatgga tcttggagga atcagatgtg cagtcaggga ccatagcaca
121 agacaggcgt cttctactgg tgctaccagc aaatgctgga agccgggaac actgggtacg
181 ttggaaacca cgtgatgtga agaagtaaga taaactgtag gagaaaagca tttcgtagtg
241 ggccatgaag cctttcagga catgtattgc agtatgggcc ggcccattac gcaattggac
301 gacaacaaag actagtatta gtaccacctc ggctatccac atagatcaaa gctgatttaa
361 aagagttgtg cagatgatcc gtggcaggag accgaggtct cggttttaga gctagaaata
421 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
481 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
541 atgcctgcag gtccccagat tagccttttc aatttcagaa agaatgctaa cccacagatg
601 gttagagagg cttacgcagc agcactcatc aagacgatct acccgagcaa taatctccag
661 gaaatcaaat accttcccaa gaaggttaaa gatgcagtca aaagattcag gactaactgc
721 atcaagaaca cagagaaaga tatatttctc aagatcagaa gtactattcc agtatggacg
781 attcaaggct tgcttcacaa accaaggcaa gtaatagaga ttggagtctc taaaaaggta
841 gttcccactg aatcaaaggc catggagtca aagattcaaa tagaggacct aacagaactc
901 gccgtaaaga ctggcgaaca gttcatacag agtctcttac gactcaatga caagaagaaa
961 atcttcgtca acatggtgga gcacgacaca cttgtctact ccaaaaatat caaagataca
1021 gtctcagaag accaaagggc aattgagact tttcaacaaa gggtaatatc cggaaacctc
1081 ctcggattcc attgcccagc tatctgtcac tttattgtga agatagtgga aaaggaaggt
1141 ggctcctaca aatgccatca ttgcgataaa ggaaaggcca tcgttgaaga tgcctctgcc
1201 gacagtggtc ccaaagatgg acccccaccc acgaggagca tcgtggaaaa agaagacgtt
1261 ccaaccacgt cttcaaagca agtggattga tgtgatatct ccactgacgt aagggatgac
1321 gcacaatccc actatccttc gcaagaccct tcctctatat aaggaagttc atttcatttg
1381 gagagaacac gggggactct agagttatca acaagtttgt acaaaaaagc aggcttcgaa
1441 ggagatagaa ccaattctct aaggaaatac ttaaccatgg actataagga ccacgacgga
1501 gactacaagg atcatgatat tgattacaaa gacgatgacg ataagatggc cccaaagaag
1561 aagcggaagg tcggtatcca cggagtccca gcagccgaca agaagtacag catcggcctg
1621 gacatcggca ccaactctgt gggctgggcc gtgatcaccg acgagtacaa ggtgcccagc
1681 aagaaattca aggtgctggg caacaccgac cggcacagca tcaagaagaa cctgatcgga
1741 gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc ggctgaagag aaccgccaga
1801 agaagataca ccagacggaa gaaccggatc tgctatctgc aagagatctt cagcaacgag
1861 atggccaagg tggacgacag cttcttccac agactggaag agtccttcct ggtggaagag
1921 gataagaagc acgagcggca ccccatcttc ggcaacatcg tggacgaggt ggcctaccac
1981 gagaagtacc ccaccatcta ccacctgaga aagaaactgg tggacagcac cgacaaggcc
2041 gacctgcggc tgatctatct ggccctggcc cacatgatca agttccgggg ccacttcctg
2101 atcgagggcg acctgaaccc cgacaacagc gacgtggaca agctgttcat ccagctggtg
2161 cagacctaca accagctgtt cgaggaaaac cccatcaacg ccagcggcgt ggacgccaag
2221 gccatcctgt ctgccagact gagcaagagc agacggctgg aaaatctgat cgcccagctg
2281 cccggcgaga agaagaatgg cctgttcgga aacctgattg ccctgagcct gggcctgacc
2341 cccaacttca agagcaactt cgacctggcc gaggatgcca aactgcagct gagcaaggac
2401 acctacgacg acgacctgga caacctgctg gcccagatcg gcgaccagta cgccgacctg
2461 tttctggccg ccaagaacct gtccgacgcc atcctgctga gcgacatcct gagagtgaac
2521 accgagatca ccaaggcccc cctgagcgcc tctatgatca agagatacga cgagcaccac
2581 caggacctga ccctgctgaa agctctcgtg cggcagcagc tgcctgagaa gtacaaagag
2641 attttcttcg accagagcaa gaacggctac gccggctaca ttgacggcgg agccagccag
2701 gaagagttct acaagttcat caagcccatc ctggaaaaga tggacggcac cgaggaactg
2761 ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc ggaccttcga caacggcagc
2821 atcccccacc agatccacct gggagagctg cacgccattc tgcggcggca ggaagatttt
2881 tacccattcc tgaaggacaa ccgggaaaag atcgagaaga tcctgacctt ccgcatcccc
2941 tactacgtgg gccctctggc caggggaaac agcagattcg cctggatgac cagaaagagc
3001 gaggaaacca tcaccccctg gaacttcgag gaagtggtgg acaagggcgc ttccgcccag
3061 agcttcatcg agcggatgac caacttcgat aagaacctgc ccaacgagaa ggtgctgccc
3121 aagcacagcc tgctgtacga gtacttcacc gtgtataacg agctgaccaa agtgaaatac
3181 gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg agcagaaaaa ggccatcgtg
3241 gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc agctgaaaga ggactacttc
3301 aagaaaatcg agtgcttcga ctccgtggaa atctccggcg tggaagatcg gttcaacgcc
3361 tccctgggca cataccacga tctgctgaaa attatcaagg acaaggactt cctggacaat
3421 gaggaaaacg aggacattct ggaagatatc gtgctgaccc tgacactgtt tgaggacaga
3481 gagatgatcg aggaacggct gaaaacctat gcccacctgt tcgacgacaa agtgatgaag
3541 cagctgaagc ggcggagata caccggctgg ggcaggctga gccggaagct gatcaacggc
3601 atccgggaca agcagtccgg caagacaatc ctggatttcc tgaagtccga cggcttcgcc
3661 aacagaaact tcatgcagct gatccacgac gacagcctga cctttaaaga ggacatccag
3721 aaagcccagg tgtccggcca gggcgatagc ctgcacgagc acattgccaa tctggccggc
3781 agccccgcca ttaagaaggg catcctgcag acagtgaagg tggtggacga gctcgtgaaa
3841 gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa tggccagaga gaaccagacc
3901 acccagaagg gacagaagaa cagccgcgag agaatgaagc ggatcgaaga gggcatcaaa
3961 gagctgggca gccagatcct gaaagaacac cccgtggaaa acacccagct gcagaacgag
4021 aagctgtacc tgtactacct gcagaatggg cgggatatgt acgtggacca ggaactggac
4081 atcaaccggc tgtccgacta cgatgtggac catatcgtgc ctcagagctt tctgaaggac
4141 gactccatcg acaacaaggt gctgaccaga agcgacaaga accggggcaa gagcgacaac
4201 gtgccctccg aagaggtcgt gaagaagatg aagaactact ggcggcagct gctgaacgcc
4261 aagctgatta cccagagaaa gttcgacaat ctgaccaagg ccgagagagg cggcctgagc
4321 gaactggata aggccggctt catcaagaga cagctggtgg aaacccggca gatcacaaag
4381 cacgtggcac agatcctgga ctcccggatg aacactaagt acgacgagaa tgacaagctg
4441 atccgggaag tgaaagtgat caccctgaag tccaagctgg tgtccgattt ccggaaggat
4501 ttccagtttt acaaagtgcg cgagatcaac aactaccacc acgcccacga cgcctacctg
4561 aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta agctggaaag cgagttcgtg
4621 tacggcgact acaaggtgta cgacgtgcgg aagatgatcg ccaagagcga gcaggaaatc
4681 ggcaaggcta ccgccaagta cttcttctac agcaacatca tgaacttttt caagaccgag
4741 attaccctgg ccaacggcga gatccggaag cggcctctga tcgagacaaa cggcgaaacc
4801 ggggagatcg tgtgggataa gggccgggat tttgccaccg tgcggaaagt gctgagcatg
4861 ccccaagtga atatcgtgaa aaagaccgag gtgcagacag gcggcttcag caaagagtct
4921 atcctgccca agaggaacag cgataagctg atcgccagaa agaaggactg ggaccctaag
4981 aagtacggcg gcttcgacag ccccaccgtg gcctattctg tgctggtggt ggccaaagtg
5041 gaaaagggca agtccaagaa actgaagagt gtgaaagagc tgctggggat caccatcatg
5101 gaaagaagca gcttcgagaa gaatcccatc gactttctgg aagccaaggg ctacaaagaa
5161 gtgaaaaagg acctgatcat caagctgcct aagtactccc tgttcgagct ggaaaacggc
5221 cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg gaaacgaact ggccctgccc
5281 tccaaatatg tgaacttcct gtacctggcc agccactatg agaagctgaa gggctccccc
5341 gaggataatg agcagaaaca gctgtttgtg gaacagcaca agcactacct ggacgagatc
5401 atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg ccgacgctaa tctggacaaa
5461 gtgctgtccg cctacaacaa gcaccgggat aagcccatca gagagcaggc cgagaatatc
5521 atccacctgt ttaccctgac caatctggga gcccctgccg ccttcaagta ctttgacacc
5581 accatcgacc ggaagaggta caccagcacc aaagaggtgc tggacgccac cctgatccac
5641 cagagcatca ccggcctgta cgagacacgg atcgacctgt ctcagctggg aggcgacaaa
5701 aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga aaaagtaaga attcgcggcc
5761 gcactcgaga tatctagacc cagctttctt gtacaaagtg gttgataaca gcgactacaa
5821 ggatgacgat gacaaggctt agagctcgaa tttccccgat cgttcaaaca tttggcaata
5881 aagtttctta agattgaatc ctgttgccgg tcttgcgatg attatcatat aatttctgtt
5941 gaattacgtt aagcatgtaa taattaacat gtaatgcatg acgttattta tgagatgggt
6001 ttttatgatt agagtcccgc aattatacat ttaatacgcg atagaaaaca aaatatagcg
6061 cgcaaactag gataaattat cgcgcgcggt gtcatctatg ttactagatc gggaattcac
6121 tggccgtcgt tttacactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt
6181 tacccaactt aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga
6241 ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gctagagcag
6301 cttgagcttg gatcagattg tcgtttcccg ccttcagttt aaactatcag tgtttgacag
6361 gatatattgg cgggtaaacc taagagaaaa gagcgtttat tagaataacg gatatttaaa
6421 agggcgtgaa aaggtttatc cgttcgtcca tttgtatgtg catgccaacc acagggttcc
6481 cctcgggatc aaagtacttt gatccaaccc ctccgctgct atagtgcagt cggcttctga
6541 cgttcagtgc agccgtcttc tgaaaacgac atgtcgcaca agtcctaagt tacgcgacag
6601 gctgccgccc tgcccttttc ctggcgtttt cttgtcgcgt gttttagtcg cataaagtag
6661 aatacttgcg actagaaccg gagacattac gccatgaaca agagcgccgc cgctggcctg
6721 ctgggctatg cccgcgtcag caccgacgac caggacttga ccaaccaacg ggccgaactg
6781 cacgcggccg gctgcaccaa gctgttttcc gagaagatca ccggcaccag gcgcgaccgc
6841 ccggagctgg ccaggatgct tgaccaccta cgccctggcg acgttgtgac agtgaccagg
6901 ctagaccgcc tggcccgcag cacccgcgac ctactggaca ttgccgagcg catccaggag
6961 gccggcgcgg gcctgcgtag cctggcagag ccgtgggccg acaccaccac gccggccggc
7021 cgcatggtgt tgaccgtgtt cgccggcatt gccgagttcg agcgttccct aatcatcgac
7081 cgcacccgga gcgggcgcga ggccgccaag gcccgaggcg tgaagtttgg cccccgccct
7141 accctcaccc cggcacagat cgcgcacgcc cgcgagctga tcgaccagga aggccgcacc
7201 gtgaaagagg cggctgcact gcttggcgtg catcgctcga ccctgtaccg cgcacttgag
7261 cgcagcgagg aagtgacgcc caccgaggcc aggcggcgcg gtgccttccg tgaggacgca
7321 ttgaccgagg ccgacgccct ggcggccgcc gagaatgaac gccaagagga acaagcatga
7381 aaccgcacca ggacggccag gacgaaccgt ttttcattac cgaagagatc gaggcggaga
7441 tgatcgcggc cgggtacgtg ttcgagccgc ccgcgcacgt ctcaaccgtg cggctgcatg
7501 aaatcctggc cggtttgtct gatgccaagc tggcggcctg gccggccagc ttggccgctg
7561 aagaaaccga gcgccgccgt ctaaaaaggt gatgtgtatt tgagtaaaac agcttgcgtc
7621 atgcggtcgc tgcgtatatg atgcgatgag taaataaaca aatacgcaag gggaacgcat
7681 gaaggttatc gctgtactta accagaaagg cgggtcaggc aagacgacca tcgcaaccca
7741 tctagcccgc gccctgcaac tcgccggggc cgatgttctg ttagtcgatt ccgatcccca
7801 gggcagtgcc cgcgattggg cggccgtgcg ggaagatcaa ccgctaaccg ttgtcggcat
7861 cgaccgcccg acgattgacc gcgacgtgaa ggccatcggc cggcgcgact tcgtagtgat
7921 cgacggagcg ccccaggcgg cggacttggc tgtgtccgcg atcaaggcag ccgacttcgt
7981 gctgattccg gtgcagccaa gcccttacga catatgggcc accgccgacc tggtggagct
8041 ggttaagcag cgcattgagg tcacggatgg aaggctacaa gcggcctttg tcgtgtcgcg
8101 ggcgatcaaa ggcacgcgca tcggcggtga ggttgccgag gcgctggccg ggtacgagct
8161 gcccattctt gagtcccgta tcacgcagcg cgtgagctac ccaggcactg ccgccgccgg
8221 cacaaccgtt cttgaatcag aacccgaggg cgacgctgcc cgcgaggtcc aggcgctggc
8281 cgctgaaatt aaatcaaaac tcatttgagt taatgaggta aagagaaaat gagcaaaagc
8341 acaaacacgc taagtgccgg ccgtccgagc gcacgcagca gcaaggctgc aacgttggcc
8401 agcctggcag acacgccagc catgaagcgg gtcaactttc agttgccggc ggaggatcac
8461 accaagctga agatgtacgc ggtacgccaa ggcaagacca ttaccgagct gctatctgaa
8521 tacatcgcgc agctaccaga gtaaatgagc aaatgaataa atgagtagat gaattttagc
8581 ggctaaagga ggcggcatgg aaaatcaaga acaaccaggc accgacgccg tggaatgccc
8641 catgtgtgga ggaacgggcg gttggccagg cgtaagcggc tgggttgtct gccggccctg
8701 caatggcact ggaaccccca agcccgagga atcggcgtga cggtcgcaaa ccatccggcc
8761 cggtacaaat cggcgcggcg ctgggtgatg acctggtgga gaagttgaag gccgcgcagg
8821 ccgcccagcg gcaacgcatc gaggcagaag cacgccccgg tgaatcgtgg caagcggccg
8881 ctgatcgaat ccgcaaagaa tcccggcaac cgccggcagc cggtgcgccg tcgattagga
8941 agccgcccaa gggcgacgag caaccagatt ttttcgttcc gatgctctat gacgtgggca
9001 cccgcgatag tcgcagcatc atggacgtgg ccgttttccg tctgtcgaag cgtgaccgac
9061 gagctggcga ggtgatccgc tacgagcttc cagacgggca cgtagaggtt tccgcagggc
9121 cggccggcat ggccagtgtg tgggattacg acctggtact gatggcggtt tcccatctaa
9181 ccgaatccat gaaccgatac cgggaaggga agggagacaa gcccggccgc gtgttccgtc
9241 cacacgttgc ggacgtactc aagttctgcc ggcgagccga tggcggaaag cagaaagacg
9301 acctggtaga aacctgcatt cggttaaaca ccacgcacgt tgccatgcag cgtacgaaga
9361 aggccaagaa cggccgcctg gtgacggtat ccgagggtga agccttgatt agccgctaca
9421 agatcgtaaa gagcgaaacc gggcggccgg agtacatcga gatcgagcta gctgattgga
9481 tgtaccgcga gatcacagaa ggcaagaacc cggacgtgct gacggttcac cccgattact
9541 ttttgatcga tcccggcatc ggccgttttc tctaccgcct ggcacgccgc gccgcaggca
9601 aggcagaagc cagatggttg ttcaagacga tctacgaacg cagtggcagc gccggagagt
9661 tcaagaagtt ctgtttcacc gtgcgcaagc tgatcgggtc aaatgacctg ccggagtacg
9721 atttgaagga ggaggcgggg caggctggcc cgatcctagt catgcgctac cgcaacctga
9781 tcgagggcga agcatccgcc ggttcctaat gtacggagca gatgctaggg caaattgccc
9841 tagcagggga aaaaggtcga aaagcactct ttcctgtgga tagcacgtac attgggaacc
9901 caaagccgta cattgggaac cggaacccgt acattgggaa cccaaagccg tacattggga
9961 accggtcaca catgtaagtg actgatataa aagagaaaaa aggcgatttt tccgcctaaa
10021 actctttaaa acttattaaa actcttaaaa cccgcctggc ctgtgcataa ctgtctggcc
10081 agcgcacagc cgaagagctg caaaaagcgc ctacccttcg gtcgctgcgc tccctacgcc
10141 ccgccgcttc gcgtcggcct atcgcggccg ctggccgctc aaaaatggct ggcctacggc
10201 caggcaatct accagggcgc ggacaagccg cgccgtcgcc actcgaccgc cggcgcccac
10261 atcaaggcac cctgcctcgc gcgtttcggt gatgacggtg aaaacctctg acacatgcag
10321 ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag
10381 ggcgcgtcag cgggtgttgg cgggtgtcgg ggcgcagcca tgacccagtc acgtagcgat
10441 agcggagtgt atactggctt aactatgcgg catcagagca gattgtactg agagtgcacc
10501 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggcgctctt
10561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
10621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
10681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
10741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
10801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
10861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
10921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
10981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
11041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
11101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
11161 actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct
11221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
11281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
11341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
11401 tgcattctag gtactaaaac aattcatcca gtaaaatata atattttatt ttctcccaat
11461 caggcttgat ccccagtaag tcaaaaaata gctcgacata ctgttcttcc ccgatatcct
11521 ccctgatcga ccggacgcag aaggcaatgt cataccactt gtccgccctg ccgcttctcc
11581 caagatcaat aaagccactt actttgccat ctttcacaaa gatgttgctg tctcccaggt
11641 cgccgtggga aaagacaagt tcctcttcgg gcttttccgt ctttaaaaaa tcatacagct
11701 cgcgcggatc tttaaatgga gtgtcttctt cccagttttc gcaatccaca tcggccagat
11761 cgttattcag taagtaatcc aattcggcta agcggctgtc taagctattc gtatagggac
11821 aatccgatat gtcgatggag tgaaagagcc tgatgcactc cgcatacagc tcgataatct
11881 tttcagggct ttgttcatct tcatactctt ccgagcaaag gacgccatcg gcctcactca
11941 tgagcagatt gctccagcca tcatgccgtt caaagtgcag gacctttgga acaggcagct
12001 ttccttccag ccatagcatc atgtcctttt cccgttccac atcataggtg gtccctttat
12061 accggctgtc cgtcattttt aaatataggt tttcattttc tcccaccagc ttatatacct
12121 tagcaggaga cattccttcc gtatctttta cgcagcggta tttttcgatc agttttttca
12181 attccggtga tattctcatt ttagccattt attatttcct tcctcttttc tacagtattt
12241 aaagataccc caagaagcta attataacaa gacgaactcc aattcactgt tccttgcatt
12301 ctaaaacctt aaataccaga aaacagcttt ttcaaagttg ttttcaaagt tggcgtataa
12361 catagtatcg acggagccga ttttgaaacc gcggtgatca caggcagcaa cgctctgtca
12421 tcgttacaat caacatgcta ccctccgcga gatcatccgt gtttcaaacc cggcagctta
12481 gttgccgttc ttccgaatag catcggtaac atgagcaaag tctgccgcct tacaacggct
12541 ctcccgctga cgccgtcccg gactgatggg ctgcctgtat cgagtggtga ttttgtgccg
12601 agctgccggt cggggagctg ttggctggct ggtggcagga tatattgtgg tgtaaacaaa
12661 ttgacgctta gacaacttaa taacacattg cggacgtttt taatgtactg aattaacgcc
12721 gaattaattc gggggatctg gattttagta ctggattttg gttttaggaa ttagaaattt
12781 tattgataga agtattttac aaatacaaat acatactaag ggtttcttat atgctcaaca
12841 catgagcgaa accctatagg aaccctaatt cccttatctg ggaactactc acacattatt
12901 atggagaaac tcgagcttgt cgatcgacag atccggtcgg catctactct atttctttgc
12961 cctcggacga gtgctggggc gtcggtttcc actatcggcg agtacttcta cacagccatc
13021 ggtccagacg gccgcgcttc tgcgggcgat ttgtgtacgc ccgacagtcc cggctccgga
13081 tcggacgatt gcgtcgcatc gaccctgcgc ccaagctgca tcatcgaaat tgccgtcaac
13141 caagctctga tagagttggt caagaccaat gcggagcata tacgcccgga gtcgtggcga
13201 tcctgcaagc tccggatgcc tccgctcgaa gtagcgcgtc tgctgctcca tacaagccaa
13261 ccacggcctc cagaagaaga tgttggcgac ctcgtattgg gaatccccga acatcgcctc
13321 gctccagtca atgaccgctg ttatgcggcc attgtccgtc aggacattgt tggagccgaa
13381 atccgcgtgc acgaggtgcc ggacttcggg gcagtcctcg gcccaaagca tcagctcatc
13441 gagagcctgc gcgacggacg cactgacggt gtcgtccatc acagtttgcc agtgatacac
13501 atggggatca gcaatcgcgc atatgaaatc acgccatgta gtgtattgac cgattccttg
13561 cggtccgaat gggccgaacc cgctcgtctg gctaagatcg gccgcagcga tcgcatccat
13621 agcctccgcg accggttgta gaacagcggg cagttcggtt tcaggcaggt cttgcaacgt
13681 gacaccctgt gcacggcggg agatgcaata ggtcaggctc tcgctaaact ccccaatgtc
13741 aagcacttcc ggaatcggga gcgcggccga tgcaaagtgc cgataaacat aacgatcttt
13801 gtagaaacca tcggcgcagc tatttacccg caggacatat ccacgccctc ctacatcgaa
13861 gctgaaagca cgagattctt cgccctccga gagctgcatc aggtcggaga cgctgtcgaa
13921 cttttcgatc agaaacttct cgacagacgt cgcggtgagt tcaggctttt tcatatctca
13981 ttgccccccg ggatctgcga aagctcgaga gagatagatt tgtagagaga gactggtgat
14041 ttcagcgtgt cctctccaaa tgaaatgaac ttccttatat agaggaaggt cttgcgaagg
14101 atagtgggat tgtgcgtcat cccttacgtc agtggagata tcacatcaat ccacttgctt
14161 tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg ggggtccatc
14221 tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat cgcaatgatg
14281 gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac agatagctgg
14341 gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct caatagccct
14401 ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt cgtgctccac
14461 catgttatca catcaatcca cttgctttga agacgtggtt ggaacgtctt ctttttccac
14521 gatgctcctc gtgggtgggg gtccatcttt gggaccactg tcggcagagg catcttgaac
14581 gatagccttt cctttatcgc aatgatggca tttgtaggtg ccaccttcct tttctactgt
14641 ccttttgatg aagtgacaga tagctgggca atggaatccg aggaggtttc ccgatattac
14701 cctttgttga aaagtctcaa tagccctttg gtcttctgag actgtatctt tgatattctt
14761 ggagtagacg agagtgtcgt gctccaccat gttggcaagc tgctctagcc aatacgcaaa
14821 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
14881 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
14941 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
15001 tttcacacag gaaacagcta tgaccatgat tacgcca
//
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