| 质粒名称: | pYLCRISPR/Cas9P35S-N |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66191 |
| 存储实验室: | Yao-Guang Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCAMBIA1300 |
| 载体出品公司: | CAMBIA |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pYLCRISPR/Cas9P35S-N was a gift from Yao-Guang Liu (Addgene plasmid # 66191 ; http://n2t.net/addgene:66191 ; RRID:Addgene_66191) |
| 参考文献部分: | A robust CRISPR/Cas9 system for convenient, high-efficiency multiplex genome editing in monocot and dicot plants. Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H, Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X, Dong Z, Liu YG. Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi: 10.1016/j.molp.2015.04.007. 10.1016/j.molp.2015.04.007 PubMed 25917172 |
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 pYLCRISPR/Cas9P35S-N 16211 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION expression of Cas9 in plants, cauliflower mosaic virus 35S promoter
(P35S), Neo selection.
ACCESSION .
VERSION .
KEYWORDS pYLCRISPR/Cas9P35S-N
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16211)
AUTHORS Ma X, Zhang Q, Zhu Q, Liu W, Chen Y, Qiu R, Wang B, Yang Z, Li H,
Lin Y, Xie Y, Shen R, Chen S, Wang Z, Chen Y, Guo J, Chen L, Zhao X,
Dong Z, Liu YG
TITLE A robust CRISPR/Cas9 system for convenient, high-efficiency
multiplex genome editing in monocot and dicot plants.
JOURNAL Mol Plant. 2015 Apr 24. pii: S1674-2052(15)00204-X. doi:
10.1016/j.molp.2015.04.007.
PUBMED 25917172
REFERENCE 2 (bases 1 to 16211)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..16211
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 260..604
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 753..773
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 798..4898
/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"
RBS 1984..1992
/label=Shine-Dalgarno sequence
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 4899..4946
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(5287..5303)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5287..5303)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5300..5322)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5311..5327
/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(5335..5365)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5380..5401
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5592..6268
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 6331..7128
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGR
PVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLL
SSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEH
QGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDI
ALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(6388..6407)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 6998..7017
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 7185..7359
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 7437..7461
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 9085..9879
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 9966..10554
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10455..10474
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10708..10725
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 10740..10880
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(10806..10825)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 10966..10988
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 11224..11418
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(11484..12557)
/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(12986..13615)
/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 14916..14940
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 15128..15150
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 15142..15159
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 15143..15159
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 15189..15222
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
CDS complement(15432..15734)
/codon_start=1
/gene="ccdB"
/product="CcdB, a bacterial toxin that poisons DNA gyrase"
/label=ccdB
/note="Plasmids containing the ccdB gene cannot be
propagated in standard E. coli strains."
/translation="QFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDKV
SRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
primer_bind complement(15534..15553)
/label=ccdB-fwd
/note="ccdB gene, forward primer"
promoter complement(15808..15826)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(15809..15826)
/label=SP6
/note="SP6 promoter, forward primer"
protein_bind 15868..15884
/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(15892..15922)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 15937..15958
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 ttattgtgaa gatagtggaa aaggaaggtg gctcctacaa atgccatcat tgcgataaag
61 gaaaggccat cgttgaagat gcctctgccg acagtggtcc caaagatgga cccccaccca
121 cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa gtggattgat
181 gtgataacat ggtggagcac gacacacttg tctactccaa aaatatcaaa gatacagtct
241 cagaagacca aagggcaatt gagacttttc aacaaagggt aatatccgga aacctcctcg
301 gattccattg cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct
361 cctacaaatg ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca
421 gtggtcccaa agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa
481 ccacgtcttc aaagcaagtg gattgatgtg atatctccac tgacgtaagg gatgacgcac
541 aatcccacta tccttcgcaa gacccttcct ctatataagg aagttcattt catttggaga
601 ggacgtcgag agttctcaac acaacatata caaaacaaac gaatctcaag caatcaagca
661 ttctacttct attgcagcaa tttaaatcat ttcttttaaa gcaaaagcaa ttttctgaaa
721 attttcacca tttacgaacg atagccatgg ctcctaagaa gaagcggaag gttggtattc
781 acggggtgcc tgcggctgac aagaagtact ccatcggcct cgacatcggc accaacagcg
841 tcggctgggc ggtgatcacc gacgagtaca aggtcccgtc caagaagttc aaggtcctgg
901 gcaacaccga ccgccactcc atcaagaaga acctcatcgg cgccctcctc ttcgactccg
961 gcgagacggc ggaggcgacc cgcctcaagc gcaccgcccg ccgccgctac acccgccgca
1021 agaaccgcat ctgctacctc caggagatct tctccaacga gatggcgaag gtcgacgact
1081 ccttcttcca ccgcctcgag gagtccttcc tcgtggagga ggacaagaag cacgagcgcc
1141 accccatctt cggcaacatc gtcgacgagg tcgcctacca cgagaagtac cccactatct
1201 accaccttcg taagaagctt gttgactcta ctgataaggc tgatcttcgt ctcatctacc
1261 ttgctctcgc tcacatgatc aagttccgtg gtcacttcct tatcgagggt gaccttaacc
1321 ctgataactc cgacgtggac aagctcttca tccagctcgt ccagacctac aaccagctct
1381 tcgaggagaa ccctatcaac gcttccggtg tcgacgctaa ggcgatcctt tccgctaggc
1441 tctccaagtc caggcgtctc gagaacctca tcgcccagct ccctggtgag aagaagaacg
1501 gtcttttcgg taacctcatc gctctctccc tcggtctgac ccctaacttc aagtccaact
1561 tcgacctcgc tgaggacgct aagcttcagc tctccaagga tacctacgac gatgatctcg
1621 acaacctcct cgctcagatt ggagatcagt acgctgatct cttccttgct gctaagaacc
1681 tctccgatgc tatcctcctt tcggatatcc ttagggttaa cactgagatc actaaggctc
1741 ctctttctgc ttccatgatc aagcgctacg acgagcacca ccaggacctc accctcctca
1801 aggctcttgt tcgtcagcag ctccccgaga agtacaagga gatcttcttc gaccagtcca
1861 agaacggcta cgccggttac attgacggtg gagctagcca ggaggagttc tacaagttca
1921 tcaagccaat ccttgagaag atggatggta ctgaggagct tctcgttaag cttaaccgtg
1981 aggacctcct taggaagcag aggactttcg ataacggctc tatccctcac cagatccacc
2041 ttggtgagct tcacgccatc cttcgtaggc aggaggactt ctaccctttc ctcaaggaca
2101 accgtgagaa gatcgagaag atccttactt tccgtattcc ttactacgtt ggtcctcttg
2161 ctcgtggtaa ctcccgtttc gcttggatga ctaggaagtc cgaggagact atcacccctt
2221 ggaacttcga ggaggttgtt gacaagggtg cttccgccca gtccttcatc gagcgcatga
2281 ccaacttcga caagaacctc cccaacgaga aggtcctccc caagcactcc ctcctctacg
2341 agtacttcac ggtctacaac gagctcacca aggtcaagta cgtcaccgag ggtatgcgca
2401 agcctgcctt cctctccggc gagcagaaga aggctatcgt tgacctcctc ttcaagacca
2461 accgcaaggt caccgtcaag cagctcaagg aggactactt caagaagatc gagtgcttcg
2521 actccgtcga gatcagcggc gttgaggacc gtttcaacgc ttctctcggt acctaccacg
2581 atctcctcaa gatcatcaag gacaaggact tcctcgacaa cgaggagaac gaggacatcc
2641 tcgaggacat cgtcctcact cttactctct tcgaggatag ggagatgatc gaggagaggc
2701 tcaagactta cgctcatctc ttcgatgaca aggttatgaa gcagctcaag cgtcgccgtt
2761 acaccggttg gggtaggctc tcccgcaagc tcatcaacgg tatcagggat aagcagagcg
2821 gcaagactat cctcgacttc ctcaagtctg atggtttcgc taacaggaac ttcatgcagc
2881 tcatccacga tgactctctt accttcaagg aggatattca gaaggctcag gtgtccggtc
2941 agggcgactc tctccacgag cacattgcta accttgctgg ttcccctgct atcaagaagg
3001 gcatccttca gactgttaag gttgtcgatg agcttgtcaa ggttatgggt cgtcacaagc
3061 ctgagaacat cgtcatcgag atggctcgtg agaaccagac tacccagaag ggtcagaaga
3121 actcgaggga gcgcatgaag aggattgagg agggtatcaa ggagcttggt tctcagatcc
3181 ttaaggagca ccctgtcgag aacacccagc tccagaacga gaagctctac ctctactacc
3241 tccagaacgg tagggatatg tacgttgacc aggagctcga catcaacagg ctttctgact
3301 acgacgtcga ccacattgtt cctcagtctt tccttaagga tgactccatc gacaacaagg
3361 tcctcacgag gtccgacaag aacaggggta agtcggacaa cgtcccttcc gaggaggttg
3421 tcaagaagat gaagaactac tggaggcagc ttctcaacgc taagctcatt acccagagga
3481 agttcgacaa cctcacgaag gctgagaggg gtggcctttc cgagcttgac aaggctggtt
3541 tcatcaagag gcagcttgtt gagacgaggc agattaccaa gcacgttgct cagatcctcg
3601 attctaggat gaacaccaag tacgacgaga acgacaagct catccgcgag gtcaaggtga
3661 tcaccctcaa gtccaagctc gtctccgact tccgcaagga cttccagttc tacaaggtcc
3721 gcgagatcaa caactaccac cacgctcacg atgcttacct taacgctgtc gttggtaccg
3781 ctcttatcaa gaagtaccct aagcttgagt ccgagttcgt ctacggtgac tacaaggtct
3841 acgacgttcg taagatgatc gccaagtccg agcaggagat cggcaaggcc accgccaagt
3901 acttcttcta ctccaacatc atgaacttct tcaagaccga gatcaccctc gccaacggcg
3961 agatccgcaa gcgccctctt atcgagacga acggtgagac tggtgagatc gtttgggaca
4021 agggtcgcga cttcgctact gttcgcaagg tcctttctat gcctcaggtt aacatcgtca
4081 agaagaccga ggtccagacc ggtggcttct ccaaggagtc tatccttcca aagagaaact
4141 cggacaagct catcgctagg aagaaggatt gggaccctaa gaagtacggt ggtttcgact
4201 cccctactgt cgcctactcc gtcctcgtgg tcgccaaggt ggagaagggt aagtcgaaga
4261 agctcaagtc cgtcaaggag ctcctcggca tcaccatcat ggagcgctcc tccttcgaga
4321 agaacccgat cgacttcctc gaggccaagg gctacaagga ggtcaagaag gacctcatca
4381 tcaagctccc caagtactct cttttcgagc tcgagaacgg tcgtaagagg atgctggctt
4441 ccgctggtga gctccagaag ggtaacgagc ttgctcttcc ttccaagtac gtgaacttcc
4501 tctacctcgc ctcccactac gagaagctca agggttcccc tgaggataac gagcagaagc
4561 agctcttcgt ggagcagcac aagcactacc tcgacgagat catcgagcag atctccgagt
4621 tctccaagcg cgtcatcctc gctgacgcta acctcgacaa ggtcctctcc gcctacaaca
4681 agcaccgcga caagcccatc cgcgagcagg ccgagaacat catccacctc ttcacgctca
4741 cgaacctcgg cgcccctgct gctttcaagt acttcgacac caccatcgac aggaagcgtt
4801 acacgtccac caaggaggtt ctcgacgcta ctctcatcca ccagtccatc accggtcttt
4861 acgagactcg tatcgacctt tcccagcttg gtggtgataa gcgtcctgct gccaccaaaa
4921 aggccggaca ggctaagaaa aagaagtagg atccacctga tctagagtcc gcaaaaatca
4981 ccagtctctc tctacaaatc tatctctctc tatttttctc cagaataatg tgtgagtagt
5041 tcccagataa gggaattagg gttcttatag ggtttcgctc atgtgttgag catataagaa
5101 acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc taattcctaa
5161 aaccaaaatc cagtgacgcg gccgcataag gccggccttg ccctagttcc tatagcctac
5221 attataggat ggagggatat cctctcttaa ggtagcgagc ggtaccgagc tcgaattcgt
5281 aatcatgtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
5341 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
5401 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
5461 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt ggctagagca gcttgccaac
5521 atggtggagc acgacactct cgtctactcc aagaatatca aagatacagt ctcagaagac
5581 caaagggcta ttgagacttt tcaacaaagg gtaatatcgg gaaacctcct cggattccat
5641 tgcccagcta tctgtcactt catcaaaagg acagtagaaa aggaaggtgg cacctacaaa
5701 tgccatcatt gcgataaagg aaaggctatc gttcaagatg cctctgccga cagtggtccc
5761 aaagatggac ccccacccac gaggagcatc gtggaaaaag aagacgttcc aaccacgtct
5821 tcaaagcaag tggattgatg tgaacatggt ggagcacgac actctcgtct actccaagaa
5881 tatcaaagat acagtctcag aagaccaaag ggctattgag acttttcaac aaagggtaat
5941 atcgggaaac ctcctcggat tccattgccc agctatctgt cacttcatca aaaggacagt
6001 agaaaaggaa ggtggcacct acaaatgcca tcattgcgat aaaggaaagg ctatcgttca
6061 agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgaaga gcatcgtgga
6121 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
6181 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
6241 ttcatttcat ttggagagga cacgctgaaa tcaccagtct ctctctacaa atctatctct
6301 ctcgagcttt cgcagatctg tcgatcgacc atggggattg aacaagatgg attgcacgca
6361 ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc
6421 ggctgctctg atgccgccgt gtttcggctg tcagcgcagg ggcgcccggt tctttttgtc
6481 aagaccgacc tgtccggtgc cctgaatgaa ctccaggacg aggcagcgcg gctatcgtgg
6541 ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg
6601 gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct
6661 gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct
6721 acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa
6781 gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa
6841 ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt gacacatggc
6901 gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt
6961 ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct
7021 gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc
7081 gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg
7141 ggttcggatc gatcctctag ctagagtcga tcgacaagct cgagtttctc cataataatg
7201 tgtgagtagt tcccagataa gggaattagg gttcctatag ggtttcgctc atgtgttgag
7261 catataagaa acccttagta tgtatttgta tttgtaaaat acttctatca ataaaatttc
7321 taattcctaa aaccaaaatc cagtactaaa atccagatcc cccgaattaa ttcggcgtta
7381 attcagtaca ttaaaaacgt ccgcaatgtg ttattaagtt gtctaagcgt caatttgttt
7441 acaccacaat atatcctgcc accagccagc caacagctcc ccgaccggca gctcggcaca
7501 aaatcaccac tcgatacagg cagcccatca gtccgggacg gcgtcagcgg gagagccgtt
7561 gtaaggcggc agactttgct catgttaccg atgctattcg gaagaacggc aactaagctg
7621 ccgggtttga aacacggatg atctcgcgga gggtagcatg ttgattgtaa cgatgacaga
7681 gcgttgctgc ctgtgatcac cgcggtttca aaatcggctc cgtcgatact atgttatacg
7741 ccaactttga aaacaacttt gaaaaagctg ttttctggta tttaaggttt tagaatgcaa
7801 ggaacagtga attggagttc gtcttgttat aattagggaa ggtgcgaata agcggggaaa
7861 ttcttctcgg ctgactcagt catttcattt cttcatgttt gagccgattt tttctcccgt
7921 aaatgccttg aatcagccta tttagaccgt ttcttcgcca tttaaggcgt tatccccagt
7981 ttttagtgag atctctccca ctgacgtatc atttggtccg cccgaaacag gttggccagc
8041 gtgaataaca tcgccagttg gttatcgttt ttcagcaacc ccttgtatct ggctttcacg
8101 aagccgaact gtcgcttgat gatgcgaaat gggtgctcca ccctggcccg gatgctggct
8161 ttcatgtatt cgatgttgat ggccgttttg ttcttgcgtg gatgctgttt caaggttctt
8221 accttgccgg ggcgctcggc gatcagccag tccacatcca cctcggccag ctcctcgcgc
8281 tgtggcgccc cttggtagcc ggcatcggct gagacaaatt gctcctctcc atgcagcaga
8341 ttacccagct gattgaggtc atgctcgttg gccgcggtgg tgaccaggct gtgggtcagg
8401 ccactcttgg catcgacacc aatgtgggcc ttcatgccaa agtgccactg attgcctttc
8461 ttggtctgat gcatctccgg atcgcgttgc tgctctttgt tcttggtcga gctgggtgcc
8521 tcaatgatgg tggcatcgac caaggtgcct tgagtcatca tgacgcctgc ttcggccagc
8581 cagcgattga tggtcttgaa caattggcgg gccagttgat gctgctccag caggtggcgg
8641 aaattcatga tggtggtgcg gtccggcaag gcgctatcca gggataaccg ggcaaacaga
8701 cgcatggagg cgatttcgta cagagcatct tccatcgcgc catcgctcag gttgtaccaa
8761 tgctgcatgc agtgaatgcg tagcatggtt tccagcggat aaggtcgccg gccattacca
8821 gccttggggt aaaacggctc gatgacttcc accatgtttt gccatggcag aatctgctcc
8881 atgcgggaca agaaaatctc ttttctggtc tgacggcgct tactgctgaa ttcactgtcg
8941 gcgaaggtaa gttgatgact catgatgaac cctgttctat ggctccagat gacaaacatg
9001 atctcatatc agggacttgt tcgcaccttc cttagcttct tggggtatct ttaaatactg
9061 tagaaaagag gaaggaaata ataaatggct aaaatgagaa tatcaccgga attgaaaaaa
9121 ctgatcgaaa aataccgctg cgtaaaagat acggaaggaa tgtctcctgc taaggtatat
9181 aagctggtgg gagaaaatga aaacctatat ttaaaaatga cggacagccg gtataaaggg
9241 accacctatg atgtggaacg ggaaaaggac atgatgctat ggctggaagg aaagctgcct
9301 gttccaaagg tcctgcactt tgaacggcat gatggctgga gcaatctgct catgagtgag
9361 gccgatggcg tcctttgctc ggaagagtat gaagatgaac aaagccctga aaagattatc
9421 gagctgtatg cggagtgcat caggctcttt cactccatcg acatatcgga ttgtccctat
9481 acgaatagct tagacagccg cttagccgaa ttggattact tactgaataa cgatctggcc
9541 gatgtggatt gcgaaaactg ggaagaagac actccattta aagatccgcg cgagctgtat
9601 gattttttaa agacggaaaa gcccgaagag gaacttgtct tttcccacgg cgacctggga
9661 gacagcaaca tctttgtgaa agatggcaaa gtaagtggct ttattgatct tgggagaagc
9721 ggcagggcgg acaagtggta tgacattgcc ttctgcgtcc ggtcgatcag ggaggatatc
9781 ggggaagaac agtatgtcga gctatttttt gacttactgg ggatcaagcc tgattgggag
9841 aaaataaaat attatatttt actggatgaa ttgttttagt acctagaatg catgaccaaa
9901 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
9961 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
10021 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
10081 ggcttcagca gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac
10141 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
10201 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg
10261 gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga
10321 acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc
10381 gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg
10441 agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc
10501 tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc
10561 agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt
10621 cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc
10681 gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc
10741 ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact
10801 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atacactccg ctatcgctac
10861 gtgactgggt catggctgcg ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
10921 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
10981 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgaggca gggtgccttg atgtgggcgc
11041 cggcggtcga gtggcgacgg cgcggcttgt ccgcgccctg gtagattgcc tggccgtagg
11101 ccagccattt ttgagcggcc agcggccgcg ataggccgac gcgaagcggc ggggcgtagg
11161 gagcgcagcg accgaagggt aggcgctttt tgcagctctt cggctgtgcg ctggccagac
11221 agttatgcac aggccaggcg ggttttaaga gttttaataa gttttaaaga gttttaggcg
11281 gaaaaatcgc cttttttctc ttttatatca gtcacttaca tgtgtgaccg gttcccaatg
11341 tacggctttg ggttcccaat gtacgggttc cggttcccaa tgtacggctt tgggttccca
11401 atgtacgtgc tatccacagg aaagagaact tttcgacctt tttcccctgc tagggcaatt
11461 tgccctagca tctgctccgt acattaggaa ccggcggatg cttcgccctc gatcaggttg
11521 cggtagcgca tgactaggat cgggccagcc tgccccgcct cctccttcaa atcgtactcc
11581 ggcaggtcat ttgacccgat cagcttgcgc acggtgaaac agaacttctt gaactctccg
11641 gcgctgccac tgcgttcgta gatcgtcttg aacaaccatc tggcttctgc cttgcctgcg
11701 gcgcggcgtg ccaggcggta gagaaaacgg ccgatgccgg gatcgatcaa aaagtaatcg
11761 gggtgaaccg tcagcacgtc cgggttcttg ccttctgtga tctcgcggta catccaatca
11821 gctagctcga tctcgatgta ctccggccgc ccggtttcgc tctttacgat cttgtagcgg
11881 ctaatcaagg cttcaccctc ggataccgtc accaggcggc cgttcttggc cttcttcgta
11941 cgctgcatgg caacgtgcgt ggtgtttaac cgaatgcagg tttctaccag gtcgtctttc
12001 tgctttccgc catcggctcg ccggcagaac ttgagtacgt ccgcaacgtg tggacggaac
12061 acgcggccgg gcttgtctcc cttcccttcc cggtatcggt tcatggattc ggttagatgg
12121 gaaaccgcca tcagtaccag gtcgtaatcc cacacactgg ccatgccggc cggccctgcg
12181 gaaacctcta cgtgcccgtc tggaagctcg tagcggatca cctcgccagc tcgtcggtca
12241 cgcttcgaca gacggaaaac ggccacgtcc atgatgctgc gactatcgcg ggtgcccacg
12301 tcatagagca tcggaacgaa aaaatctggt tgctcgtcgc ccttgggcgg cttcctaatc
12361 gacggcgcac cggctgccgg cggttgccgg gattctttgc ggattcgatc agcggccgct
12421 tgccacgatt caccggggcg tgcttctgcc tcgatgcgtt gccgctgggc ggcctgcgcg
12481 gccttcaact tctccaccag gtcatcaccc agcgccgcgc cgatttgtac cgggccggat
12541 ggtttgcgac cgctcacgcc gattcctcgg gcttgggggt tccagtgcca ttgcagggcc
12601 ggcaggcaac ccagccgctt acgcctggcc aaccgcccgt tcctccacac atggggcatt
12661 ccacggcgtc ggtgcctggt tgttcttgat tttccatgcc gcctccttta gccgctaaaa
12721 ttcatctact catttattca tttgctcatt tactctggta gctgcgcgat gtattcagat
12781 agcagctcgg taatggtctt gccttggcgt accgcgtaca tcttcagctt ggtgtgatcc
12841 tccgccggca actgaaagtt gacccgcttc atggctggcg tgtctgccag gctggccaac
12901 gttgcagcct tgctgctgcg tgcgctcgga cggccggcac ttagcgtgtt tgtgcttttg
12961 ctcattttct ctttacctca ttaactcaaa tgagttttga tttaatttca gcggccagcg
13021 cctggacctc gcgggcagcg tcgccctcgg gttctgattc aagaacggtt gtgccggcgg
13081 cggcagtgcc tgggtagctc acgcgctgcg tgatacggga ctcaagaatg ggcagctcgt
13141 acccggccag cgcctcggca acctcaccgc cgatgcgcgt gcctttgatc gcccgcgaca
13201 cgacaaaggc cgcttgtagc cttccatccg tgacctcaat gcgctgctta accagctcca
13261 ccaggtcggc ggtggcccat atgtcgtaag ggcttggctg caccggaatc agcacgaagt
13321 cggctgcctt gatcgcggac acagccaagt ccgccgcctg gggcgctccg tcgatcacta
13381 cgaagtcgcg ccggccgatg gccttcacgt cgcggtcaat cgtcgggcgg tcgatgccga
13441 caacggttag cggttgatct tcccgcacgg ccgcccaatc gcgggcactg ccctggggat
13501 cggaatcgac taacagaaca tcggccccgg cgagttgcag ggcgcgggct agatgggttg
13561 cgatggtcgt cttgcctgac ccgcctttct ggttaagtac agcgataacc ttcatgcgtt
13621 ccccttgcgt atttgtttat ttactcatcg catcatatac gcagcgaccg catgacgcaa
13681 gctgttttac tcaaatacac atcacctttt tagacggcgg cgctcggttt cttcagcggc
13741 caagctggcc ggccaggccg ccagcttggc atcagacaaa ccggccagga tttcatgcag
13801 ccgcacggtt gagacgtgcg cgggcggctc gaacacgtac ccggccgcga tcatctccgc
13861 ctcgatctct tcggtaatga aaaacggttc gtcctggccg tcctggtgcg gtttcatgct
13921 tgttcctctt ggcgttcatt ctcggcggcc gccagggcgt cggcctcggt caatgcgtcc
13981 tcacggaagg caccgcgccg cctggcctcg gtgggcgtca cttcctcgct gcgctcaagt
14041 gcgcggtaca gggtcgagcg atgcacgcca agcagtgcag ccgcctcttt cacggtgcgg
14101 ccttcctggt cgatcagctc gcgggcgtgc gcgatctgtg ccggggtgag ggtagggcgg
14161 gggccaaact tcacgcctcg ggccttggcg gcctcgcgcc cgctccgggt gcggtcgatg
14221 attagggaac gctcgaactc ggcaatgccg gcgaacacgg tcaacaccat gcggccggcc
14281 ggcgtggtgg tgtcggccca cggctctgcc aggctacgca ggcccgcgcc ggcctcctgg
14341 atgcgctcgg caatgtccag taggtcgcgg gtgctgcggg ccaggcggtc tagcctggtc
14401 actgtcacaa cgtcgccagg gcgtaggtgg tcaagcatcc tggccagctc cgggcggtcg
14461 cgcctggtgc cggtgatctt ctcggaaaac agcttggtgc agccggccgc gtgcagttcg
14521 gcccgttggt tggtcaagtc ctggtcgtcg gtgctgacgc gggcatagcc cagcaggcca
14581 gcggcggcgc tcttgttcat ggcgtaatgt ctccggttct agtcgcaagt attctacttt
14641 atgcgactaa aacacgcgac aagaaaacgc caggaaaagg gcagggcggc agcctgtcgc
14701 gtaacttagg acttgtgcga catgtcgttt tcagaagacg gctgcactga acgtcagaag
14761 ccgactgcac tatagcagcg gaggggttgg atcaaagtac tttgatcccg aggggaaccc
14821 tgtggttggc atgcacatac aaatggacga acggataaac cttttcacgc ccttttaaat
14881 atccgattat tctaataaac gctcttttct cttaggttta cccgccaata tatcctgtca
14941 aacactgata gtttaaactg aaggcgggaa acgacaatct gatccaagct caagctgctc
15001 tagcattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
15061 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
15121 gggttttccc agtcacgacg ttgtaaaacg acggccagtg ccaagcttat gggtgtcgac
15181 ttgcagtcat aacttcgtat agcatacatt atacgaagtt atcagatcta atactgccat
15241 ttcattacct ctttctccgc acccgacata gatgcaataa cttcgtatag gctaatttat
15301 acgatgtagg agatcgatgc atgcggccgc tagctcgaga ggcgcgccaa tgataccgcg
15361 agacccacgc tcacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
15421 aggcctgaca tttatattcc ccagaacatc aggttaatgg cgtttttgat gtcattttcg
15481 cggtggctga gatcagccac ttcttccccg ataacggaga caggcacact ggccatatcg
15541 gtggtcatca tgcgccagct ttcatccccg atatgcacca ccgggtaaag ttcacgggag
15601 actttatctg acagcagacg tgcactggcc agggggatca ccatccgtcg ccccggcgtg
15661 tcaataatat cactctgtac atccacaaac agacgataac ggctctctct tttataggtg
15721 taaaccttaa actgatgttc gagcggtcgc cagtgtgatg ggtatctaca gactagaccc
15781 attgagcttg atgcatagct tgagtattct atagtgtcac ctaaatagct tggcgtaatc
15841 atggtcatag ctgtttcttg tgtgaaattg ttatccgctc acaattccac acaacatacg
15901 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
15961 cgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
16021 aatcggccac ttcagaggtc tctcgagcac tacggcgcgc cttaccggtc aacatgtgga
16081 gcacgacaca cttgtctact ccaaaaatat caaagataca gtctcagaag accaaagggc
16141 aattgagact tttcaacaaa gggtaatatc cggaaacctc ctcggattcc attgcccagc
16201 tatctgtcac t
//
