| 质粒名称: | pYLCRISPR/Cas9Pubi-H |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 66187 |
| 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: | 卡那霉素 |
| 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: | 植物表达 |
| 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: | -- |
| 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/Cas9Pubi-H was a gift from Yao-Guang Liu (Addgene plasmid # 66187 ; http://n2t.net/addgene:66187 ; RRID:Addgene_66187) |
| 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/Cas9Pubi-H 17619 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION expression of Cas9 in plants, maize ubiquitin promoter (Pubi), hygro
selection.
ACCESSION .
VERSION .
KEYWORDS pYLCRISPR/Cas9Pubi-H
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17619)
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 17619)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..17619
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 730..750
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 775..4875
/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 1961..1969
/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 4876..4923
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
terminator 4936..5188
/label=NOS terminator
/note="nopaline synthase terminator and poly(A) signal"
protein_bind 5306..5327
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 5342..5372
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5380..5396
/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 5438..5456
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind 5438..5455
/label=SP6
/note="SP6 promoter, forward primer"
CDS 5530..5832
/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 5711..5730
/label=ccdB-fwd
/note="ccdB gene, forward primer"
protein_bind complement(6042..6075)
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
primer_bind complement(6119..6136)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(6119..6135)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6128..6150)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_feature 6338..6362
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
CDS 7663..8292
/codon_start=1
/product="stability protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 StaA
/translation="MKVIAVLNQKGGSGKTTIATHLARALQLAGADVLLVDSDPQGSAR
DWAAVREDQPLTVVGIDRPTIDRDVKAIGRRDFVVIDGAPQAADLAVSAIKAADFVLIP
VQPSPYDIWATADLVELVKQRIEVTDGRLQAAFVVSRAIKGTRIGGEVAEALAGYELPI
LESRITQRVSYPGTAAAGTTVLESEPEGDAAREVQALAAEIKSKLI"
CDS 8721..9794
/codon_start=1
/product="replication protein from the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
/label=pVS1 RepA
/translation="MSGRKPSGPVQIGAALGDDLVEKLKAAQAAQRQRIEAEARPGESW
QAAADRIRKESRQPPAAGAPSIRKPPKGDEQPDFFVPMLYDVGTRDSRSIMDVAVFRLS
KRDRRAGEVIRYELPDGHVEVSAGPAGMASVWDYDLVLMAVSHLTESMNRYREGKGDKP
GRVFRPHVADVLKFCRRADGGKQKDDLVETCIRLNTTHVAMQRTKKAKNGRLVTVSEGE
ALISRYKIVKSETGRPEYIEIELADWMYREITEGKNPDVLTVHPDYFLIDPGIGRFLYR
LARRAAGKAEARWLFKTIYERSGSAGEFKKFCFTVRKLIGSNDLPEYDLKEEAGQAGPI
LVMRYRNLIEGEASAGS"
rep_origin 9860..10054
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
primer_bind complement(10290..10312)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
misc_feature 10398..10538
/label=bom
/note="basis of mobility region from pBR322"
primer_bind 10453..10472
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(10553..10570)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10724..11312)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(10804..10823)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(11399..12193)
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
misc_feature 13817..13841
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
polyA_signal 13919..14093
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
CDS complement(14134..15159)
/codon_start=1
/gene="aph(4)-Ia"
/product="aminoglycoside phosphotransferase from E. coli"
/label=HygR
/note="confers resistance to hygromycin"
/translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRSQGVTLQDLP
ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
K"
promoter complement(15226..15902)
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind complement(15269..15289)
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
protein_bind 16093..16114
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 16129..16159
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 16167..16183
/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 16172..16194
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 16191..16207
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 16191..16207
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
ORIGIN
1 ttgccctttt cctttatttc aatatatgcc gtgcacttgt ttgtcgggtc atcttttcat
61 gctttttttt gtcttggttg tgatgatgtg gtctggttgg gcggtcgttc tagatcggag
121 tagaattctg tttcaaacta cctggtggat ttattaattt tggatctgta tgtgtgtgcc
181 atacatattc atagttacga attgaagatg atggatggaa atatcgatct aggataggta
241 tacatgttga tgcgggtttt actgatgcat atacagagat gctttttgtt cgcttggttg
301 tgatgatgtg gtgtggttgg gcggtcgttc attcgttcta gatcggagta gaatactgtt
361 tcaaactacc tggtgtattt attaattttg gaactgtatg tgtgtgtcat acatcttcat
421 agttacgagt ttaagatgga tggaaatatc gatctaggat aggtatacat gttgatgtgg
481 gttttactga tgcatataca tgatggcata tgcagcatct attcatatgc tctaaccttg
541 agtacctatc tattataata aacaagtatg ttttataatt attttgatct tgatatactt
601 ggatgatggc atatgcagca gctatatgtg gattttttta gccctgcctt catacgctat
661 ttatttgctt ggtactgttt cttttgtcga tgctcaccct gttgtttggt gttacttctg
721 cagatggctc ctaagaagaa gcggaaggtt ggtattcacg gggtgcctgc ggctgacaag
781 aagtactcca tcggcctcga catcggcacc aacagcgtcg gctgggcggt gatcaccgac
841 gagtacaagg tcccgtccaa gaagttcaag gtcctgggca acaccgaccg ccactccatc
901 aagaagaacc tcatcggcgc cctcctcttc gactccggcg agacggcgga ggcgacccgc
961 ctcaagcgca ccgcccgccg ccgctacacc cgccgcaaga accgcatctg ctacctccag
1021 gagatcttct ccaacgagat ggcgaaggtc gacgactcct tcttccaccg cctcgaggag
1081 tccttcctcg tggaggagga caagaagcac gagcgccacc ccatcttcgg caacatcgtc
1141 gacgaggtcg cctaccacga gaagtacccc actatctacc accttcgtaa gaagcttgtt
1201 gactctactg ataaggctga tcttcgtctc atctaccttg ctctcgctca catgatcaag
1261 ttccgtggtc acttccttat cgagggtgac cttaaccctg ataactccga cgtggacaag
1321 ctcttcatcc agctcgtcca gacctacaac cagctcttcg aggagaaccc tatcaacgct
1381 tccggtgtcg acgctaaggc gatcctttcc gctaggctct ccaagtccag gcgtctcgag
1441 aacctcatcg cccagctccc tggtgagaag aagaacggtc ttttcggtaa cctcatcgct
1501 ctctccctcg gtctgacccc taacttcaag tccaacttcg acctcgctga ggacgctaag
1561 cttcagctct ccaaggatac ctacgacgat gatctcgaca acctcctcgc tcagattgga
1621 gatcagtacg ctgatctctt ccttgctgct aagaacctct ccgatgctat cctcctttcg
1681 gatatcctta gggttaacac tgagatcact aaggctcctc tttctgcttc catgatcaag
1741 cgctacgacg agcaccacca ggacctcacc ctcctcaagg ctcttgttcg tcagcagctc
1801 cccgagaagt acaaggagat cttcttcgac cagtccaaga acggctacgc cggttacatt
1861 gacggtggag ctagccagga ggagttctac aagttcatca agccaatcct tgagaagatg
1921 gatggtactg aggagcttct cgttaagctt aaccgtgagg acctccttag gaagcagagg
1981 actttcgata acggctctat ccctcaccag atccaccttg gtgagcttca cgccatcctt
2041 cgtaggcagg aggacttcta ccctttcctc aaggacaacc gtgagaagat cgagaagatc
2101 cttactttcc gtattcctta ctacgttggt cctcttgctc gtggtaactc ccgtttcgct
2161 tggatgacta ggaagtccga ggagactatc accccttgga acttcgagga ggttgttgac
2221 aagggtgctt ccgcccagtc cttcatcgag cgcatgacca acttcgacaa gaacctcccc
2281 aacgagaagg tcctccccaa gcactccctc ctctacgagt acttcacggt ctacaacgag
2341 ctcaccaagg tcaagtacgt caccgagggt atgcgcaagc ctgccttcct ctccggcgag
2401 cagaagaagg ctatcgttga cctcctcttc aagaccaacc gcaaggtcac cgtcaagcag
2461 ctcaaggagg actacttcaa gaagatcgag tgcttcgact ccgtcgagat cagcggcgtt
2521 gaggaccgtt tcaacgcttc tctcggtacc taccacgatc tcctcaagat catcaaggac
2581 aaggacttcc tcgacaacga ggagaacgag gacatcctcg aggacatcgt cctcactctt
2641 actctcttcg aggataggga gatgatcgag gagaggctca agacttacgc tcatctcttc
2701 gatgacaagg ttatgaagca gctcaagcgt cgccgttaca ccggttgggg taggctctcc
2761 cgcaagctca tcaacggtat cagggataag cagagcggca agactatcct cgacttcctc
2821 aagtctgatg gtttcgctaa caggaacttc atgcagctca tccacgatga ctctcttacc
2881 ttcaaggagg atattcagaa ggctcaggtg tccggtcagg gcgactctct ccacgagcac
2941 attgctaacc ttgctggttc ccctgctatc aagaagggca tccttcagac tgttaaggtt
3001 gtcgatgagc ttgtcaaggt tatgggtcgt cacaagcctg agaacatcgt catcgagatg
3061 gctcgtgaga accagactac ccagaagggt cagaagaact cgagggagcg catgaagagg
3121 attgaggagg gtatcaagga gcttggttct cagatcctta aggagcaccc tgtcgagaac
3181 acccagctcc agaacgagaa gctctacctc tactacctcc agaacggtag ggatatgtac
3241 gttgaccagg agctcgacat caacaggctt tctgactacg acgtcgacca cattgttcct
3301 cagtctttcc ttaaggatga ctccatcgac aacaaggtcc tcacgaggtc cgacaagaac
3361 aggggtaagt cggacaacgt cccttccgag gaggttgtca agaagatgaa gaactactgg
3421 aggcagcttc tcaacgctaa gctcattacc cagaggaagt tcgacaacct cacgaaggct
3481 gagaggggtg gcctttccga gcttgacaag gctggtttca tcaagaggca gcttgttgag
3541 acgaggcaga ttaccaagca cgttgctcag atcctcgatt ctaggatgaa caccaagtac
3601 gacgagaacg acaagctcat ccgcgaggtc aaggtgatca ccctcaagtc caagctcgtc
3661 tccgacttcc gcaaggactt ccagttctac aaggtccgcg agatcaacaa ctaccaccac
3721 gctcacgatg cttaccttaa cgctgtcgtt ggtaccgctc ttatcaagaa gtaccctaag
3781 cttgagtccg agttcgtcta cggtgactac aaggtctacg acgttcgtaa gatgatcgcc
3841 aagtccgagc aggagatcgg caaggccacc gccaagtact tcttctactc caacatcatg
3901 aacttcttca agaccgagat caccctcgcc aacggcgaga tccgcaagcg ccctcttatc
3961 gagacgaacg gtgagactgg tgagatcgtt tgggacaagg gtcgcgactt cgctactgtt
4021 cgcaaggtcc tttctatgcc tcaggttaac atcgtcaaga agaccgaggt ccagaccggt
4081 ggcttctcca aggagtctat ccttccaaag agaaactcgg acaagctcat cgctaggaag
4141 aaggattggg accctaagaa gtacggtggt ttcgactccc ctactgtcgc ctactccgtc
4201 ctcgtggtcg ccaaggtgga gaagggtaag tcgaagaagc tcaagtccgt caaggagctc
4261 ctcggcatca ccatcatgga gcgctcctcc ttcgagaaga acccgatcga cttcctcgag
4321 gccaagggct acaaggaggt caagaaggac ctcatcatca agctccccaa gtactctctt
4381 ttcgagctcg agaacggtcg taagaggatg ctggcttccg ctggtgagct ccagaagggt
4441 aacgagcttg ctcttccttc caagtacgtg aacttcctct acctcgcctc ccactacgag
4501 aagctcaagg gttcccctga ggataacgag cagaagcagc tcttcgtgga gcagcacaag
4561 cactacctcg acgagatcat cgagcagatc tccgagttct ccaagcgcgt catcctcgct
4621 gacgctaacc tcgacaaggt cctctccgcc tacaacaagc accgcgacaa gcccatccgc
4681 gagcaggccg agaacatcat ccacctcttc acgctcacga acctcggcgc ccctgctgct
4741 ttcaagtact tcgacaccac catcgacagg aagcgttaca cgtccaccaa ggaggttctc
4801 gacgctactc tcatccacca gtccatcacc ggtctttacg agactcgtat cgacctttcc
4861 cagcttggtg gtgataagcg tcctgctgcc accaaaaagg ccggacaggc taagaaaaag
4921 aagtaggatc ctcccgatcg ttcaaacatt tggcaataaa gtttcttaag attgaatcct
4981 gttgccggtc ttgcgatgat tatcatataa tttctgttga attacgttaa gcatgtaata
5041 attaacatgt aatgcatgac gttatttatg aggtgggttt ttatgattag agtcccgcaa
5101 ttatacattt aatacgcgat agaaaacaaa atatagcgcg caaactagga taaattatcg
5161 cgcgcggtgt catctatgtt actagatcgg gagcaccggt aaggcgcgcc gtagtgctcg
5221 agagacctct gaagtggccg attcattaat gcagctggca cgacaggttt cccgactgga
5281 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
5341 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
5401 acacaagaaa cagctatgac catgattacg ccaagctatt taggtgacac tatagaatac
5461 tcaagctatg catcaagctc aatgggtcta gtctgtagat acccatcaca ctggcgaccg
5521 ctcgaacatc agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg
5581 gatgtacaga gtgatattat tgacacgccg gggcgacgga tggtgatccc cctggccagt
5641 gcacgtctgc tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat
5701 gaaagctggc gcatgatgac caccgatatg gccagtgtgc ctgtctccgt tatcggggaa
5761 gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc
5821 tggggaatat aaatgtcagg cctgaatggc gaatggacgc gccctgtagc ggcgcattaa
5881 gcgcggcggg tgagcgtggg tctcgcggta tcattggcgc gcctctcgag ctagcggccg
5941 catgcatcga tctcctacat cgtataaatt agcctatacg aagttattgc atctatgtcg
6001 ggtgcggaga aagaggtaat gaaatggcag tattagatct gataacttcg tataatgtat
6061 gctatacgaa gttatgactg caggtcgaca cccataatag ctgtttgcca agcttggcac
6121 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
6181 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
6241 cttcccaaca gttgcgcagc ctgaatggcg aatgctagag cagcttgagc ttggatcaga
6301 ttgtcgtttc ccgccttcag tttaaactat cagtgtttga caggatatat tggcgggtaa
6361 acctaagaga aaagagcgtt tattagaata atcggatatt taaaagggcg tgaaaaggtt
6421 tatccgttcg tccatttgta tgtgcatgcc aaccacaggg ttcccctcgg gatcaaagta
6481 ctttgatcca acccctccgc tgctatagtg cagtcggctt ctgacgttca gtgcagccgt
6541 cttctgaaaa cgacatgtcg cacaagtcct aagttacgcg acaggctgcc gccctgccct
6601 tttcctggcg ttttcttgtc gcgtgtttta gtcgcataaa gtagaatact tgcgactaga
6661 accggagaca ttacgccatg aacaagagcg ccgccgctgg cctgctgggc tatgcccgcg
6721 tcagcaccga cgaccaggac ttgaccaacc aacgggccga actgcacgcg gccggctgca
6781 ccaagctgtt ttccgagaag atcaccggca ccaggcgcga ccgcccggag ctggccagga
6841 tgcttgacca cctacgccct ggcgacgttg tgacagtgac caggctagac cgcctggccc
6901 gcagcacccg cgacctactg gacattgccg agcgcatcca ggaggccggc gcgggcctgc
6961 gtagcctggc agagccgtgg gccgacacca ccacgccggc cggccgcatg gtgttgaccg
7021 tgttcgccgg cattgccgag ttcgagcgtt ccctaatcat cgaccgcacc cggagcgggc
7081 gcgaggccgc caaggcccga ggcgtgaagt ttggcccccg ccctaccctc accccggcac
7141 agatcgcgca cgcccgcgag ctgatcgacc aggaaggccg caccgtgaaa gaggcggctg
7201 cactgcttgg cgtgcatcgc tcgaccctgt accgcgcact tgagcgcagc gaggaagtga
7261 cgcccaccga ggccaggcgg cgcggtgcct tccgtgagga cgcattgacc gaggccgacg
7321 ccctggcggc cgccgagaat gaacgccaag aggaacaagc atgaaaccgc accaggacgg
7381 ccaggacgaa ccgtttttca ttaccgaaga gatcgaggcg gagatgatcg cggccgggta
7441 cgtgttcgag ccgcccgcgc acgtctcaac cgtgcggctg catgaaatcc tggccggttt
7501 gtctgatgcc aagctggcgg cctggccggc cagcttggcc gctgaagaaa ccgagcgccg
7561 ccgtctaaaa aggtgatgtg tatttgagta aaacagcttg cgtcatgcgg tcgctgcgta
7621 tatgatgcga tgagtaaata aacaaatacg caaggggaac gcatgaaggt tatcgctgta
7681 cttaaccaga aaggcgggtc aggcaagacg accatcgcaa cccatctagc ccgcgccctg
7741 caactcgccg gggccgatgt tctgttagtc gattccgatc cccagggcag tgcccgcgat
7801 tgggcggccg tgcgggaaga tcaaccgcta accgttgtcg gcatcgaccg cccgacgatt
7861 gaccgcgacg tgaaggccat cggccggcgc gacttcgtag tgatcgacgg agcgccccag
7921 gcggcggact tggctgtgtc cgcgatcaag gcagccgact tcgtgctgat tccggtgcag
7981 ccaagccctt acgacatatg ggccaccgcc gacctggtgg agctggttaa gcagcgcatt
8041 gaggtcacgg atggaaggct acaagcggcc tttgtcgtgt cgcgggcgat caaaggcacg
8101 cgcatcggcg gtgaggttgc cgaggcgctg gccgggtacg agctgcccat tcttgagtcc
8161 cgtatcacgc agcgcgtgag ctacccaggc actgccgccg ccggcacaac cgttcttgaa
8221 tcagaacccg agggcgacgc tgcccgcgag gtccaggcgc tggccgctga aattaaatca
8281 aaactcattt gagttaatga ggtaaagaga aaatgagcaa aagcacaaac acgctaagtg
8341 ccggccgtcc gagcgcacgc agcagcaagg ctgcaacgtt ggccagcctg gcagacacgc
8401 cagccatgaa gcgggtcaac tttcagttgc cggcggagga tcacaccaag ctgaagatgt
8461 acgcggtacg ccaaggcaag accattaccg agctgctatc tgaatacatc gcgcagctac
8521 cagagtaaat gagcaaatga ataaatgagt agatgaattt tagcggctaa aggaggcggc
8581 atggaaaatc aagaacaacc aggcaccgac gccgtggaat gccccatgtg tggaggaacg
8641 ggcggttggc caggcgtaag cggctgggtt gcctgccggc cctgcaatgg cactggaacc
8701 cccaagcccg aggaatcggc gtgagcggtc gcaaaccatc cggcccggta caaatcggcg
8761 cggcgctggg tgatgacctg gtggagaagt tgaaggccgc gcaggccgcc cagcggcaac
8821 gcatcgaggc agaagcacgc cccggtgaat cgtggcaagc ggccgctgat cgaatccgca
8881 aagaatcccg gcaaccgccg gcagccggtg cgccgtcgat taggaagccg cccaagggcg
8941 acgagcaacc agattttttc gttccgatgc tctatgacgt gggcacccgc gatagtcgca
9001 gcatcatgga cgtggccgtt ttccgtctgt cgaagcgtga ccgacgagct ggcgaggtga
9061 tccgctacga gcttccagac gggcacgtag aggtttccgc agggccggcc ggcatggcca
9121 gtgtgtggga ttacgacctg gtactgatgg cggtttccca tctaaccgaa tccatgaacc
9181 gataccggga agggaaggga gacaagcccg gccgcgtgtt ccgtccacac gttgcggacg
9241 tactcaagtt ctgccggcga gccgatggcg gaaagcagaa agacgacctg gtagaaacct
9301 gcattcggtt aaacaccacg cacgttgcca tgcagcgtac gaagaaggcc aagaacggcc
9361 gcctggtgac ggtatccgag ggtgaagcct tgattagccg ctacaagatc gtaaagagcg
9421 aaaccgggcg gccggagtac atcgagatcg agctagctga ttggatgtac cgcgagatca
9481 cagaaggcaa gaacccggac gtgctgacgg ttcaccccga ttactttttg atcgatcccg
9541 gcatcggccg ttttctctac cgcctggcac gccgcgccgc aggcaaggca gaagccagat
9601 ggttgttcaa gacgatctac gaacgcagtg gcagcgccgg agagttcaag aagttctgtt
9661 tcaccgtgcg caagctgatc gggtcaaatg acctgccgga gtacgatttg aaggaggagg
9721 cggggcaggc tggcccgatc ctagtcatgc gctaccgcaa cctgatcgag ggcgaagcat
9781 ccgccggttc ctaatgtacg gagcagatgc tagggcaaat tgccctagca ggggaaaaag
9841 gtcgaaaagt tctctttcct gtggatagca cgtacattgg gaacccaaag ccgtacattg
9901 ggaaccggaa cccgtacatt gggaacccaa agccgtacat tgggaaccgg tcacacatgt
9961 aagtgactga tataaaagag aaaaaaggcg atttttccgc ctaaaactct ttaaaactta
10021 ttaaaactct taaaacccgc ctggcctgtg cataactgtc tggccagcgc acagccgaag
10081 agctgcaaaa agcgcctacc cttcggtcgc tgcgctccct acgccccgcc gcttcgcgtc
10141 ggcctatcgc ggccgctggc cgctcaaaaa tggctggcct acggccaggc aatctaccag
10201 ggcgcggaca agccgcgccg tcgccactcg accgccggcg cccacatcaa ggcaccctgc
10261 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc
10321 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
10381 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
10441 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg cggtgtgaaa
10501 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
10561 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
10621 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
10681 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
10741 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
10801 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
10861 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
10921 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
10981 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
11041 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
11101 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
11161 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
11221 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
11281 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
11341 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgcat tctaggtact
11401 aaaacaattc atccagtaaa atataatatt ttattttctc ccaatcaggc ttgatcccca
11461 gtaagtcaaa aaatagctcg acatactgtt cttccccgat atcctccctg atcgaccgga
11521 cgcagaaggc aatgtcatac cacttgtccg ccctgccgct tctcccaaga tcaataaagc
11581 cacttacttt gccatctttc acaaagatgt tgctgtctcc caggtcgccg tgggaaaaga
11641 caagttcctc ttcgggcttt tccgtcttta aaaaatcata cagctcgcgc ggatctttaa
11701 atggagtgtc ttcttcccag ttttcgcaat ccacatcggc cagatcgtta ttcagtaagt
11761 aatccaattc ggctaagcgg ctgtctaagc tattcgtata gggacaatcc gatatgtcga
11821 tggagtgaaa gagcctgatg cactccgcat acagctcgat aatcttttca gggctttgtt
11881 catcttcata ctcttccgag caaaggacgc catcggcctc actcatgagc agattgctcc
11941 agccatcatg ccgttcaaag tgcaggacct ttggaacagg cagctttcct tccagccata
12001 gcatcatgtc cttttcccgt tccacatcat aggtggtccc tttataccgg ctgtccgtca
12061 tttttaaata taggttttca ttttctccca ccagcttata taccttagca ggagacattc
12121 cttccgtatc ttttacgcag cggtattttt cgatcagttt tttcaattcc ggtgatattc
12181 tcattttagc catttattat ttccttcctc ttttctacag tatttaaaga taccccaaga
12241 agctaaggaa ggtgcgaaca agtccctgat atgagatcat gtttgtcatc tggagccata
12301 gaacagggtt catcatgagt catcaactta ccttcgccga cagtgaattc agcagtaagc
12361 gccgtcagac cagaaaagag attttcttgt cccgcatgga gcagattctg ccatggcaaa
12421 acatggtgga agtcatcgag ccgttttacc ccaaggctgg taatggccgg cgaccttatc
12481 cgctggaaac catgctacgc attcactgca tgcagcattg gtacaacctg agcgatggcg
12541 cgatggaaga tgctctgtac gaaatcgcct ccatgcgtct gtttgcccgg ttatccctgg
12601 atagcgcctt gccggaccgc accaccatca tgaatttccg ccacctgctg gagcagcatc
12661 aactggcccg ccaattgttc aagaccatca atcgctggct ggccgaagca ggcgtcatga
12721 tgactcaagg caccttggtc gatgccacca tcattgaggc acccagctcg accaagaaca
12781 aagagcagca acgcgatccg gagatgcatc agaccaagaa aggcaatcag tggcactttg
12841 gcatgaaggc ccacattggt gtcgatgcca agagtggcct gacccacagc ctggtcacca
12901 ccgcggccaa cgagcatgac ctcaatcagc tgggtaatct gctgcatgga gaggagcaat
12961 ttgtctcagc cgatgccggc taccaagggg cgccacagcg cgaggagctg gccgaggtgg
13021 atgtggactg gctgatcgcc gagcgccccg gcaaggtaag aaccttgaaa cagcatccac
13081 gcaagaacaa aacggccatc aacatcgaat acatgaaagc cagcatccgg gccagggtgg
13141 agcacccatt tcgcatcatc aagcgacagt tcggcttcgt gaaagccaga tacaaggggt
13201 tgctgaaaaa cgataaccaa ctggcgatgt tattcacgct ggccaacctg tttcgggcgg
13261 accaaatgat acgtcagtgg gagagatctc actaaaaact ggggataacg ccttaaatgg
13321 cgaagaaacg gtctaaatag gctgattcaa ggcatttacg ggagaaaaaa tcggctcaaa
13381 catgaagaaa tgaaatgact gagtcagccg agaagaattt ccccgcttat tcgcaccttc
13441 cctaattata acaagacgaa ctccaattca ctgttccttg cattctaaaa ccttaaatac
13501 cagaaaacag ctttttcaaa gttgttttca aagttggcgt ataacatagt atcgacggag
13561 ccgattttga aaccgcggtg atcacaggca gcaacgctct gtcatcgtta caatcaacat
13621 gctaccctcc gcgagatcat ccgtgtttca aacccggcag cttagttgcc gttcttccga
13681 atagcatcgg taacatgagc aaagtctgcc gccttacaac ggctctcccg ctgacgccgt
13741 cccggactga tgggctgcct gtatcgagtg gtgattttgt gccgagctgc cggtcgggga
13801 gctgttggct ggctggtggc aggatatatt gtggtgtaaa caaattgacg cttagacaac
13861 ttaataacac attgcggacg tttttaatgt actgaattaa cgccgaatta attcggggga
13921 tctggatttt agtactggat tttggtttta ggaattagaa attttattga tagaagtatt
13981 ttacaaatac aaatacatac taagggtttc ttatatgctc aacacatgag cgaaacccta
14041 taggaaccct aattccctta tctgggaact actcacacat tattatggag aaactcgagc
14101 ttgtcgatcg acagatcccg gtcggcatct actctatttc tttgccctcg gacgagtgct
14161 ggggcgtcgg tttccactat cggcgagtac ttctacacag ccatcggtcc agacggccgc
14221 gcttctgcgg gcgatttgtg tacgcccgac agtcccggct ccggatcgga cgattgcgtc
14281 gcatcgaccc tgcgcccaag ctgcatcatc gaaattgccg tcaaccaagc tctgatagag
14341 ttggtcaaga ccaatgcgga gcatatacgc ccggagtcgt ggcgatcctg caagctccgg
14401 atgcctccgc tcgaagtagc gcgtctgctg ctccatacaa gccaaccacg gcctccagaa
14461 gaagatgttg gcgacctcgt attgggaatc cccgaacatc gcctcgctcc agtcaatgac
14521 cgctgttatg cggccattgt ccgtcaggac attgttggag ccgaaatccg cgtgcacgag
14581 gtgccggact tcggggcagt cctcggccca aagcatcagc tcatcgagag cctgcgcgac
14641 ggacgcactg acggtgtcgt ccatcacagt ttgccagtga tacacatggg gatcagcaat
14701 cgcgcatatg aaatcacgcc atgtagtgta ttgaccgatt ccttgcggtc cgaatgggcc
14761 gaacccgctc gtctggctaa gatcggccgc agcgatcgca tccatagcct ccgcgaccgg
14821 ttgtagaaca gcgggcagtt cggtttcagg caggtcttgc aacgtgacac cctgtgaacg
14881 gcgggagatg caataggtca ggctctcgct aaactcccca atgtcaagca cttccggaat
14941 cgggagcgcg gccgatgcaa agtgccgata aacataacga tctttgtaga aaccatcggc
15001 gcagctattt acccgcagga catatccacg ccctcctaca tcgaagctga aagcacgaga
15061 ttcttcgccc tccgagagct gcatcaggtc ggagacgctg tcgaactttt cgatcagaaa
15121 cttctcgaca gacgtcgcgg tgagttcagg ctttttcata tctcattgcc ccccggatct
15181 gcgaaagctc gagagagata gatttgtaga gagagactgg tgatttcagc gtgtcctctc
15241 caaatgaaat gaacttcctt atatagagga agggtcttgc gaaggatagt gggattgtgc
15301 gtcatccctt acgtcagtgg agatatcaca tcaatccact tgctttgaag acgtggttgg
15361 aacgtcttct ttttccacga tgctcttcgt gggtgggggt ccatctttgg gaccactgtc
15421 ggcagaggca tcttgaacga tagcctttcc tttatcgcaa tgatggcatt tgtaggtgcc
15481 accttccttt tctactgtcc ttttgatgaa gtgacagata gctgggcaat ggaatccgag
15541 gaggtttccc gatattaccc tttgttgaaa agtctcaata gccctttggt cttctgagac
15601 tgtatctttg atattcttgg agtagacgag agtgtcgtgc tccaccatgt tcacatcaat
15661 ccacttgctt tgaagacgtg gttggaacgt cttctttttc cacgatgctc ctcgtgggtg
15721 ggggtccatc tttgggacca ctgtcggcag aggcatcttg aacgatagcc tttcctttat
15781 cgcaatgatg gcatttgtag gtgccacctt ccttttctac tgtccttttg atgaagtgac
15841 agatagctgg gcaatggaat ccgaggaggt ttcccgatat taccctttgt tgaaaagtct
15901 caatagccct ttggtcttct gagactgtat ctttgatatt cttggagtag acgagagtgt
15961 cgtgctccac catgttggca agctgctcta gccaatacgc aaaccgcctc tccccgcgcg
16021 ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag cgggcagtga
16081 gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt tacactttat
16141 gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca caggaaacag
16201 ctatgacatg attacgaatt cgagctcggt accgtaacta taacggtcct aaggtagcga
16261 aggatccgct cgctacctta agagaggata tccctccatc ctataatgta ggctatagga
16321 actagggcaa ggccggccat gcggccgcaa gctgggtgca gcgtgacccg gtcgtgcccc
16381 tctctagaga taatgagcat tgcatgtcta agttataaaa aattaccaca tatttttttt
16441 gtcacacttg tttgaagtgc agtttatcta tctttataca tatatttaaa ctttactcta
16501 cgaataatat aatctatagt actacaataa tatcagtgtt ttagagaatc atataaatga
16561 acagttagac atggtctaaa ggacaattga gtattttgac aacaggactc tacagtttta
16621 tctttttagt gtgcatgtgt tctccttttt ttttgcaaat agcttcacct atataatact
16681 tcatccattt tattagtaca tccatttagg gtttagggtt aatggttttt atagactaat
16741 ttttttagta catctatttt attctatttt agcctctaaa ttaagaaaac taaaactcta
16801 ttttagtttt tttatttaat aatttagata taaaatagaa taaaataaag tgactaaaaa
16861 ttaaacaaat accctttaag aaattaaaaa aactaaggaa acatttttct tgtttcgagt
16921 agataatgcc agcctgttaa acgccgtcga cgagtctaac ggacaccaac cagcgaacca
16981 gcagcgtcgc gtcgggccaa gcgaagcaga cggcacggca tctctgtcgc tgcctctgga
17041 cccctctcga gagttccgct ccaccgttgg acttgctccg ctgtcggcat ccagaaattg
17101 cgtggcggag cggcagacgt gagccggcac ggcaggcggc ctcctcctcc tctcacggca
17161 ccggcagcta cgggggattc ctttcccacc gctccttcgc tttcccttcc tcgcccgccg
17221 taataaatag acaccccctc cacaccctct ttccccaacc tcgtgttgtt cggagcgcac
17281 acacacacaa ccagatctcc cccaaatcca cccgtcggca cctccgcttc aaggtacgcc
17341 gctcgtcctc cccccccccc ntctctacct tctctagatc ggcgttccgg tccatggtta
17401 gggcccggta gttctacttc tgttcatgtt tgtgttagat ccgtgtttgt gttagatccg
17461 tgctgctagc gttcgtacac ggatgcgacc tgtacgtcag acacgttctg attgctaact
17521 tgccagtgtt tctctttggg gaatcctggg atggctctag ccgttccgca gacgggatcg
17581 atttcatgat tttttttgtt tcgttgcata gggtttggt
//
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