| 质粒名称: | pUB-Cas9 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 86556 |
| 存储实验室: | Andreas Weber |
|
载体骨架基本信息 |
|
| 载体名称: | pKB65 |
| 载体出品公司: | Kristin Bernhardt |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 11630 bp |
| 完整质粒大小: | 14121 bp |
| 载体修饰: | -- |
| 载体类型: | 植物表达，CRISPR |
| 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: | Cas9 |
| 物种来源: | Synthetic |
| 插入大小: | 4137 bp |
| 基因突变: | Cas9 is codon optimized for C. reinhardtii |
| GenBank编号 | -- |
| 启动子 | UBIQUITIN10 |
| 标签/融合蛋白: | -- |
| 克隆方法: | Gateway克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | TTTCGTTCGATCCCAATTTC |
| 3' 测序引物 | aagaccggcaacaggattc |
|
文献引用方法 |
|
| 材料和方法部分: | pUB-Cas9 was a gift from Andreas Weber (Addgene plasmid # 86556 ; http://n2t.net/addgene:86556 ; RRID:Addgene_86556) |
| 参考文献部分: | An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis thaliana. Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP. Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039. eCollection 2017. 10.3389/fpls.2017.00039 PubMed 28174584 |
买家导航
pUB-Cas9载体 pUBCas9质粒基本信息
pUB-Cas9质粒图谱 pUBCas9载体图谱和pUB-Cas9载体序列 pUBCas9质粒序列多克隆位点信息
pUB-Cas9质粒 pUBCas9载体简介
pUB-Cas9质粒序列 pUBCas9载体序列
LOCUS pUB-Cas9 14128 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION For ubiquitous expression of Cas9 in planta. sgRNA can be introduced
as described in Hahn et al. (2017).
ACCESSION .
VERSION .
KEYWORDS pUB-Cas9
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14128)
AUTHORS Hahn F, Mantegazza O, Greiner A, Hegemann P, Eisenhut M, Weber AP
TITLE An Efficient Visual Screen for CRISPR/Cas9 Activity in Arabidopsis
thaliana.
JOURNAL Front Plant Sci. 2017 Jan 24;8:39. doi: 10.3389/fpls.2017.00039.
eCollection 2017.
PUBMED 28174584
REFERENCE 2 (bases 1 to 14128)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14128
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(12..28)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12..28)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(25..47)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 36..52
/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(60..90)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 105..126
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 317..993
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 930..950
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 1060..2085
/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"
polyA_signal 2126..2300
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 2378..2402
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 2827..3621
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 3708..4296
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4197..4216
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 4450..4467
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 4482..4622
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(4548..4567)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 4708..4730
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 4966..5160
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(5226..6299)
/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(6728..7357)
/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 8658..8682
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 8870..8892
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 8884..8901
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 8885..8901
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 9610..9634
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 9655..9675
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 9676..13779
/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"
protein_bind complement(13783..13807)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
terminator 13871..14123
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
ORIGIN
1 ttcgtaatca tgtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
61 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
121 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
181 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattggcta gagcagcttg
241 ccaacatggt ggagcacgac actctcgtct actccaagaa tatcaaagat acagtctcag
301 aagaccaaag ggctattgag acttttcaac aaagggtaat atcgggaaac ctcctcggat
361 tccattgccc agctatctgt cacttcatca aaaggacagt agaaaaggaa ggtggcacct
421 acaaatgcca tcattgcgat aaaggaaagg ctatcgttca agatgcctct gccgacagtg
481 gtcccaaaga tggaccccca cccacgagga gcatcgtgga aaaagaagac gttccaacca
541 cgtcttcaaa gcaagtggat tgatgtgaac atggtggagc acgacactct cgtctactcc
601 aagaatatca aagatacagt ctcagaagac caaagggcta ttgagacttt tcaacaaagg
661 gtaatatcgg gaaacctcct cggattccat tgcccagcta tctgtcactt catcaaaagg
721 acagtagaaa aggaaggtgg cacctacaaa tgccatcatt gcgataaagg aaaggctatc
781 gttcaagatg cctctgccga cagtggtccc aaagatggac ccccacccac gaggagcatc
841 gtggaaaaag aagacgttcc aaccacgtct tcaaagcaag tggattgatg tgatatctcc
901 actgacgtaa gggatgacgc acaatcccac tatccttcgc aagacccttc ctctatataa
961 ggaagttcat ttcatttgga gaggacacgc tgaaatcacc agtctctctc tacaaatcta
1021 tctctctcga gctttcgcag atccgggggg caatgagata tgaaaaagcc tgaactcacc
1081 gcgacgtctg tcgagaagtt tctgatcgaa aagttcgaca gcgtctccga cctgatgcag
1141 ctctcggagg gcgaagaatc tcgtgctttc agcttcgatg taggagggcg tggatatgtc
1201 ctgcgggtaa atagctgcgc cgatggtttc tacaaagatc gttatgttta tcggcacttt
1261 gcatcggccg cgctcccgat tccggaagtg cttgacattg gggagtttag cgagagcctg
1321 acctattgca tctcccgccg ttcacagggt gtcacgttgc aagacctgcc tgaaaccgaa
1381 ctgcccgctg ttctacaacc ggtcgcggag gctatggatg cgatcgctgc ggccgatctt
1441 agccagacga gcgggttcgg cccattcgga ccgcaaggaa tcggtcaata cactacatgg
1501 cgtgatttca tatgcgcgat tgctgatccc catgtgtatc actggcaaac tgtgatggac
1561 gacaccgtca gtgcgtccgt cgcgcaggct ctcgatgagc tgatgctttg ggccgaggac
1621 tgccccgaag tccggcacct cgtgcacgcg gatttcggct ccaacaatgt cctgacggac
1681 aatggccgca taacagcggt cattgactgg agcgaggcga tgttcgggga ttcccaatac
1741 gaggtcgcca acatcttctt ctggaggccg tggttggctt gtatggagca gcagacgcgc
1801 tacttcgagc ggaggcatcc ggagcttgca ggatcgccac gactccgggc gtatatgctc
1861 cgcattggtc ttgaccaact ctatcagagc ttggttgacg gcaatttcga tgatgcagct
1921 tgggcgcagg gtcgatgcga cgcaatcgtc cgatccggag ccgggactgt cgggcgtaca
1981 caaatcgccc gcagaagcgc ggccgtctgg accgatggct gtgtagaagt actcgccgat
2041 agtggaaacc gacgccccag cactcgtccg agggcaaaga aatagagtag atgccgaccg
2101 ggatctgtcg atcgacaagc tcgagtttct ccataataat gtgtgagtag ttcccagata
2161 agggaattag ggttcctata gggtttcgct catgtgttga gcatataaga aacccttagt
2221 atgtatttgt atttgtaaaa tacttctatc aataaaattt ctaattccta aaaccaaaat
2281 ccagtactaa aatccagatc ccccgaatta attcggcgtt aattcagtac attaaaaacg
2341 tccgcaatgt gttattaagt tgtctaagcg tcaatttgtt tacaccacaa tatatcctgc
2401 caccagccag ccaacagctc cccgaccggc agctcggcac aaaatcacca ctcgatacag
2461 gcagcccatc agtccgggac ggcgtcagcg ggagagccgt tgtaaggcgg cagactttgc
2521 tcatgttacc gatgctattc ggaagaacgg caactaagct gccgggtttg aaacacggat
2581 gatctcgcgg agggtagcat gttgattgta acgatgacag agcgttgctg cctgtgatca
2641 ccgcggtttc aaaatcggct ccgtcgatac tatgttatac gccaactttg aaaacaactt
2701 tgaaaaagct gttttctggt atttaaggtt ttagaatgca aggaacagtg aattggagtt
2761 cgtcttgtta taattagctt cttggggtat ctttaaatac tgtagaaaag aggaaggaaa
2821 taataaatgg ctaaaatgag aatatcaccg gaattgaaaa aactgatcga aaaataccgc
2881 tgcgtaaaag atacggaagg aatgtctcct gctaaggtat ataagctggt gggagaaaat
2941 gaaaacctat atttaaaaat gacggacagc cggtataaag ggaccaccta tgatgtggaa
3001 cgggaaaagg acatgatgct atggctggaa ggaaagctgc ctgttccaaa ggtcctgcac
3061 tttgaacggc atgatggctg gagcaatctg ctcatgagtg aggccgatgg cgtcctttgc
3121 tcggaagagt atgaagatga acaaagccct gaaaagatta tcgagctgta tgcggagtgc
3181 atcaggctct ttcactccat cgacatatcg gattgtccct atacgaatag cttagacagc
3241 cgcttagccg aattggatta cttactgaat aacgatctgg ccgatgtgga ttgcgaaaac
3301 tgggaagaag acactccatt taaagatccg cgcgagctgt atgatttttt aaagacggaa
3361 aagcccgaag aggaacttgt cttttcccac ggcgacctgg gagacagcaa catctttgtg
3421 aaagatggca aagtaagtgg ctttattgat cttgggagaa gcggcagggc ggacaagtgg
3481 tatgacattg ccttctgcgt ccggtcgatc agggaggata tcggggaaga acagtatgtc
3541 gagctatttt ttgacttact ggggatcaag cctgattggg agaaaataaa atattatatt
3601 ttactggatg aattgtttta gtacctagaa tgcatgacca aaatccctta acgtgagttt
3661 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
3721 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
3781 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
3841 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
3901 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
3961 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
4021 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
4081 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
4141 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
4201 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
4261 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
4321 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
4381 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
4441 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
4501 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
4561 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
4621 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
4681 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
4741 catcaccgaa acgcgcgagg cagggtgcct tgatgtgggc gccggcggtc gagtggcgac
4801 ggcgcggctt gtccgcgccc tggtagattg cctggccgta ggccagccat ttttgagcgg
4861 ccagcggccg cgataggccg acgcgaagcg gcggggcgta gggagcgcag cgaccgaagg
4921 gtaggcgctt tttgcagctc ttcggctgtg cgctggccag acagttatgc acaggccagg
4981 cgggttttaa gagttttaat aagttttaaa gagttttagg cggaaaaatc gccttttttc
5041 tcttttatat cagtcactta catgtgtgac cggttcccaa tgtacggctt tgggttccca
5101 atgtacgggt tccggttccc aatgtacggc tttgggttcc caatgtacgt gctatccaca
5161 ggaaagagac cttttcgacc tttttcccct gctagggcaa tttgccctag catctgctcc
5221 gtacattagg aaccggcgga tgcttcgccc tcgatcaggt tgcggtagcg catgactagg
5281 atcgggccag cctgccccgc ctcctccttc aaatcgtact ccggcaggtc atttgacccg
5341 atcagcttgc gcacggtgaa acagaacttc ttgaactctc cggcgctgcc actgcgttcg
5401 tagatcgtct tgaacaacca tctggcttct gccttgcctg cggcgcggcg tgccaggcgg
5461 tagagaaaac ggccgatgcc gggatcgatc aaaaagtaat cggggtgaac cgtcagcacg
5521 tccgggttct tgccttctgt gatctcgcgg tacatccaat cagctagctc gatctcgatg
5581 tactccggcc gcccggtttc gctctttacg atcttgtagc ggctaatcaa ggcttcaccc
5641 tcggataccg tcaccaggcg gccgttcttg gccttcttcg tacgctgcat ggcaacgtgc
5701 gtggtgttta accgaatgca ggtttctacc aggtcgtctt tctgctttcc gccatcggct
5761 cgccggcaga acttgagtac gtccgcaacg tgtggacgga acacgcggcc gggcttgtct
5821 cccttccctt cccggtatcg gttcatggat tcggttagat gggaaaccgc catcagtacc
5881 aggtcgtaat cccacacact ggccatgccg gccggccctg cggaaacctc tacgtgcccg
5941 tctggaagct cgtagcggat cacctcgcca gctcgtcggt cacgcttcga cagacggaaa
6001 acggccacgt ccatgatgct gcgactatcg cgggtgccca cgtcatagag catcggaacg
6061 aaaaaatctg gttgctcgtc gcccttgggc ggcttcctaa tcgacggcgc accggctgcc
6121 ggcggttgcc gggattcttt gcggattcga tcagcggccg cttgccacga ttcaccgggg
6181 cgtgcttctg cctcgatgcg ttgccgctgg gcggcctgcg cggccttcaa cttctccacc
6241 aggtcatcac ccagcgccgc gccgatttgt accgggccgg atggtttgcg accgctcacg
6301 ccgattcctc gggcttgggg gttccagtgc cattgcaggg ccggcaggca acccagccgc
6361 ttacgcctgg ccaaccgccc gttcctccac acatggggca ttccacggcg tcggtgcctg
6421 gttgttcttg attttccatg ccgcctcctt tagccgctaa aattcatcta ctcatttatt
6481 catttgctca tttactctgg tagctgcgcg atgtattcag atagcagctc ggtaatggtc
6541 ttgccttggc gtaccgcgta catcttcagc ttggtgtgat cctccgccgg caactgaaag
6601 ttgacccgct tcatggctgg cgtgtctgcc aggctggcca acgttgcagc cttgctgctg
6661 cgtgcgctcg gacggccggc acttagcgtg tttgtgcttt tgctcatttt ctctttacct
6721 cattaactca aatgagtttt gatttaattt cagcggccag cgcctggacc tcgcgggcag
6781 cgtcgccctc gggttctgat tcaagaacgg ttgtgccggc ggcggcagtg cctgggtagc
6841 tcacgcgctg cgtgatacgg gactcaagaa tgggcagctc gtacccggcc agcgcctcgg
6901 caacctcacc gccgatgcgc gtgcctttga tcgcccgcga cacgacaaag gccgcttgta
6961 gccttccatc cgtgacctca atgcgctgct taaccagctc caccaggtcg gcggtggccc
7021 atatgtcgta agggcttggc tgcaccggaa tcagcacgaa gtcggctgcc ttgatcgcgg
7081 acacagccaa gtccgccgcc tggggcgctc cgtcgatcac tacgaagtcg cgccggccga
7141 tggccttcac gtcgcggtca atcgtcgggc ggtcgatgcc gacaacggtt agcggttgat
7201 cttcccgcac ggccgcccaa tcgcgggcac tgccctgggg atcggaatcg actaacagaa
7261 catcggcccc ggcgagttgc agggcgcggg ctagatgggt tgcgatggtc gtcttgcctg
7321 acccgccttt ctggttaagt acagcgataa ccttcatgcg ttccccttgc gtatttgttt
7381 atttactcat cgcatcatat acgcagcgac cgcatgacgc aagctgtttt actcaaatac
7441 acatcacctt tttagacggc ggcgctcggt ttcttcagcg gccaagctgg ccggccaggc
7501 cgccagcttg gcatcagaca aaccggccag gatttcatgc agccgcacgg ttgagacgtg
7561 cgcgggcggc tcgaacacgt acccggccgc gatcatctcc gcctcgatct cttcggtaat
7621 gaaaaacggt tcgtcctggc cgtcctggtg cggtttcatg cttgttcctc ttggcgttca
7681 ttctcggcgg ccgccagggc gtcggcctcg gtcaatgcgt cctcacggaa ggcaccgcgc
7741 cgcctggcct cggtgggcgt cacttcctcg ctgcgctcaa gtgcgcggta cagggtcgag
7801 cgatgcacgc caagcagtgc agccgcctct ttcacggtgc ggccttcctg gtcgatcagc
7861 tcgcgggcgt gcgcgatctg tgccggggtg agggtagggc gggggccaaa cttcacgcct
7921 cgggccttgg cggcctcgcg cccgctccgg gtgcggtcga tgattaggga acgctcgaac
7981 tcggcaatgc cggcgaacac ggtcaacacc atgcggccgg ccggcgtggt ggtgtcggcc
8041 cacggctctg ccaggctacg caggcccgcg ccggcctcct ggatgcgctc ggcaatgtcc
8101 agtaggtcgc gggtgctgcg ggccaggcgg tctagcctgg tcactgtcac aacgtcgcca
8161 gggcgtaggt ggtcaagcat cctggccagc tccgggcggt cgcgcctggt gccggtgatc
8221 ttctcggaaa acagcttggt gcagccggcc gcgtgcagtt cggcccgttg gttggtcaag
8281 tcctggtcgt cggtgctgac gcgggcatag cccagcaggc cagcggcggc gctcttgttc
8341 atggcgtaat gtctccggtt ctagtcgcaa gtattctact ttatgcgact aaaacacgcg
8401 acaagaaaac gccaggaaaa gggcagggcg gcagcctgtc gcgtaactta ggacttgtgc
8461 gacatgtcgt tttcagaaga cggctgcact gaacgtcaga agccgactgc actatagcag
8521 cggaggggtt ggatcaaagt actttgatcc cgaggggaac cctgtggttg gcatgcacat
8581 acaaatggac gaacggataa accttttcac gcccttttaa atatccgatt attctaataa
8641 acgctctttt ctcttaggtt tacccgccaa tatatcctgt caaacactga tagtttaaac
8701 tgaaggcggg aaacgacaat ctgatccaag ctcaagctgc tctagcattc gccattcagg
8761 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
8821 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
8881 cgttgtaaaa cgacggccag tgccaagctt gattacgaat tcgagctcgg tacccgacga
8941 gtcagtaata aacggcgtca aagtggttgc agccggcaca cacgagtcgt gtttatcaac
9001 tcaaagcaca aatacttttc ctcaacctaa aaataaggca attagccaaa aacaactttg
9061 cgtgtaaaca acgctcaata cacgtgtcat tttattatta gctattgctt caccgcctta
9121 gctttctcgt gacctagtcg tcctcgtctt ttcttcttct tcttctataa aacaataccc
9181 aaagagctct tcttcttcac aattcagatt tcaatttctc aaaatcttaa aaactttctc
9241 tcaattctct ctaccgtgat caaggtaaat ttctgtgttc cttattctct caaaatcttc
9301 gattttgttt tcgttcgatc ccaatttcgt atatgttctt tggtttagat tctgttaatc
9361 ttagatcgaa gacgattttc tgggtttgat cgttagatat catcttaatt ctcgattagg
9421 gtttcataga tatcatccga tttgttcaaa taatttgagt tttgtcgaat aattactctt
9481 cgatttgtga tttctatcta gatctggtgt tagtttctag tttgtgcgat cgaatttgta
9541 gattaatctg agtttttctg attaacagct cgagtgcggg ccccccctcg aggcgcgcca
9601 agctatcaaa caagtttgta caaaaaagca ggctaccata tggccagcag cccccccaag
9661 aagaagcgca aggtcgacaa gaagtactcc atcggcctgg acatcggcac gaacagcgtg
9721 ggctgggccg tgatcaccga cgagtacaag gtgccgtcca agaagttcaa ggtgctgggc
9781 aacaccgacc gccacagcat caagaagaac ctgatcggcg ccctgctgtt cgactcgggc
9841 gagaccgccg aggcgacgcg cctgaagcgc accgcgcgcc gccgctacac gcgccgcaag
9901 aaccgcatct gctacctgca ggagatcttc agcaacgaga tggccaaggt ggacgactcg
9961 ttcttccacc gcctggagga gtccttcctg gtggaggagg acaagaagca cgagcgccac
10021 cccatcttcg gcaacatcgt ggacgaggtg gcctaccacg agaagtaccc gacgatctac
10081 cacctgcgca agaagctggt ggacagcacc gacaaggcgg acctgcgcct gatctacctg
10141 gccctggcgc acatgatcaa gttccgcggc cacttcctga tcgagggcga cctgaacccc
10201 gacaactcgg acgtggacaa gctgttcatc cagctggtgc agacctacaa ccagctgttc
10261 gaggagaacc cgatcaacgc ctcgggcgtg gacgccaagg cgatcctgtc cgcgcgcctg
10321 tccaagagcc gccgcttgga gaacctgatc gcccagctgc ccggcgagaa gaagaacggc
10381 ctgttcggca acctgatcgc gctgagcctg ggcctgacgc cgaacttcaa gtcgaacttc
10441 gacctggccg aggacgcgaa gctgcagctg agcaaggaca cctacgacga cgacctggac
10501 aacctgctgg cccagatcgg cgaccagtac gcggacctgt tcctggccgc gaagaacctg
10561 tcggacgcca tcctgctgtc cgacatcctg cgcgtgaaca ccgagatcac gaaggccccc
10621 ctgtcggcgt ccatgatcaa gcgctacgac gagcaccacc aggacctgac cctgctgaag
10681 gcgctggtgc gccagcagct gccggagaag tacaaggaga tcttcttcga ccagtccaag
10741 aacggctacg ccggctacat cgacggcggc gccagccagg aggagttcta caagttcatc
10801 aagcccatcc tggagaagat ggacggcacg gaggagctgc tggtgaagct gaaccgcgag
10861 gacctgctgc gcaagcagcg caccttcgac aacggcagca tcccccacca gatccacctg
10921 ggcgagctgc acgccatcct gcgccgccag gaggacttct acccgttcct gaaggacaac
10981 cgcgagaaga tcgagaagat cctgacgttc cgcatcccct actacgtggg cccgctggct
11041 cgcggcaact cccgcttcgc gtggatgacc cgcaagagcg aggagaccat cacgccctgg
11101 aacttcgagg aggtggtgga caagggcgcc agcgcgcagt cgttcatcga gcgcatgacc
11161 aacttcgaca agaacctgcc caacgagaag gtgctgccga agcactccct gctgtacgag
11221 tacttcaccg tgtacaacga gctgacgaag gtgaagtacg tgaccgaggg catgcgcaag
11281 cccgccttcc tgagcggcga gcagaagaag gcgatcgtgg acctgctgtt caagaccaac
11341 cgcaaggtga cggtgaagca gctgaaggag gactacttca agaagatcga gtgcttcgac
11401 agcgtggaga tctcgggcgt ggaggaccgc ttcaacgcca gcctgggcac ctaccacgac
11461 ctgctgaaga tcatcaagga caaggacttc ctggacaacg aggagaacga ggacatcctg
11521 gaggacatcg tgctgaccct gacgctgttc gaggaccgcg agatgatcga ggagcgcctg
11581 aagacgtacg cccacctgtt cgacgacaag gtgatgaagc agctgaagcg ccgccgctac
11641 accggctggg gccgcctgag ccgcaagctg atcaacggca tccgcgacaa gcagtcgggc
11701 aagaccatcc tggacttcct gaagtccgac ggcttcgcga accgcaactt catgcagctg
11761 atccacgacg actcgctgac cttcaaggag gacatccaga aggcccaggt gtcgggccag
11821 ggcgactccc tgcacgagca catcgccaac ctggcgggct cccccgcgat caagaagggc
11881 atcctgcaga ccgtgaaggt ggtggacgag ctggtgaagg tgatgggccg ccacaagccg
11941 gagaacatcg tgatcgagat ggcccgcgag aaccagacca cgcagaaggg ccagaagaac
12001 agccgcgagc gcatgaagcg catcgaggag ggcatcaagg agctgggctc gcagatcctg
12061 aaggagcacc ccgtggagaa cacccagctg cagaacgaga agctgtacct gtactacctg
12121 cagaacggcc gcgacatgta cgtggaccag gagctggaca tcaaccgcct gtccgactac
12181 gacgtggacc acatcgtgcc ccagagcttc ctgaaggacg actcgatcga caacaaggtg
12241 ctgacccgca gcgacaagaa ccgcggcaag agcgacaacg tgccgtcgga ggaggtggtg
12301 aagaagatga agaactactg gcgccagctg ctgaacgcca agctgatcac gcagcgcaag
12361 ttcgacaacc tgaccaaggc cgagcgcggc ggcctgtcgg agctggacaa ggcgggcttc
12421 atcaagcgcc agctggtgga gacccgccag atcacgaagc acgtggcgca gatcctggac
12481 tcccgcatga acacgaagta cgacgagaac gacaagctga tccgcgaggt gaaggtgatc
12541 accctgaagt ccaagctggt cagcgacttc cgcaaggact tccagttcta caaggtgcgc
12601 gagatcaaca actaccacca cgcccacgac gcgtacctga acgccgtggt gggcaccgcg
12661 ctgatcaaga agtaccccaa gctggagagc gagttcgtgt acggcgacta caaggtgtac
12721 gacgtgcgca agatgatcgc caagtcggag caggagatcg gcaaggccac cgcgaagtac
12781 ttcttctact ccaacatcat gaacttcttc aagaccgaga tcacgctggc caacggcgag
12841 atccgcaagc gccccctgat cgagaccaac ggcgagacgg gcgagatcgt gtgggacaag
12901 ggccgcgact tcgcgaccgt gcgcaaggtg ctgagcatgc cccaggtgaa catcgtgaag
12961 aagaccgagg tgcagacggg cggcttctcc aaggagagca tcctgccgaa gcgcaactcg
13021 gacaagctga tcgcccgcaa gaaggactgg gaccccaaga agtacggcgg cttcgactcc
13081 ccgaccgtgg cctacagcgt gctggtggtg gcgaaggtgg agaagggcaa gtccaagaag
13141 ctgaagagcg tgaaggagct gctgggcatc accatcatgg agcgcagctc gttcgagaag
13201 aaccccatcg acttcctgga ggcgaagggc tacaaggagg tgaagaagga cctgatcatc
13261 aagctgccga agtactcgct gttcgagctg gagaacggcc gcaagcgcat gctggcctcc
13321 gcgggcgagc tgcagaaggg caacgagctg gccctgccca gcaagtacgt gaacttcctg
13381 tacctggcgt cccactacga gaagctgaag ggctcgccgg aggacaacga gcagaagcag
13441 ctgttcgtgg agcagcacaa gcactacctg gacgagatca tcgagcagat ctcggagttc
13501 tccaagcgcg tgatcctggc cgacgcgaac ctggacaagg tgctgagcgc ctacaacaag
13561 caccgcgaca agcccatccg cgagcaggcg gagaacatca tccacctgtt caccctgacg
13621 aacctgggcg ccccggccgc gttcaagtac ttcgacacca cgatcgaccg caagcgctac
13681 accagcacga aggaggtgct ggacgcgacc ctgatccacc agagcatcac cggcctgtac
13741 gagacgcgca tcgacctgtc gcagctgggc ggcgactaat gtacccagct ttcttgtaca
13801 aagtggttcg ataattctta attaactagt tctagagcgg ccgccaccgc ggtggagctc
13861 gaatttcccc gatcgttcaa acatttggca ataaagtttc ttaagattga atcctgttgc
13921 cggtcttgcg atgattatca tataatttct gttgaattac gttaagcatg taataattaa
13981 catgtaatgc atgacgttat ttatgagatg ggtttttatg attagagtcc cgcaattata
14041 catttaatac gcgatagaaa acaaaatata gcgcgcaaac taggataaat tatcgcgcgc
14101 ggtgtcatct atgttactag atcgggaa
//
