| 质粒名称: | p35S-PBE |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 119772 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pJIT163 |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | 4919 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 植物表达 |
| 筛选标记: | -- |
| 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: | NLS-APOBEC1-nCas9-UGI-NLS,APOBEC1-nCas9-UGI |
| 物种来源: | -- |
| 插入大小: | 5214 bp |
| 基因突变: | nCas9 has a mutation D10A in wild type SpCas9 |
| GenBank编号 | -- |
| 启动子 | CaMv 2x35S |
| 标签/融合蛋白: |
NLS（插入件上的N端子）
NLS（插接件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | p35S-PBE was a gift from Caixia Gao (Addgene plasmid # 119772 ; http://n2t.net/addgene:119772 ; RRID:Addgene_119772) |
| 参考文献部分: | Efficient C-to-T base editing in plants using a fusion of nCas9 and human APOBEC3A. Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C. Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261. 10.1038/nbt.4261 PubMed 30272679 |
买家导航
p35SPBE载体基本信息
p35SPBE载体图谱质粒图谱和多克隆位点信息
p35SPBE载体简介
p35SPBE载体序列
LOCUS p35S-PBE 17062 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Conversion of C to T in potato.
ACCESSION .
VERSION .
KEYWORDS p35S-PBE
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 17062)
AUTHORS Zong Y, Song Q, Li C, Jin S, Zhang D, Wang Y, Qiu JL, Gao C
TITLE Efficient C-to-T base editing in plants using a fusion of nCas9 and
human APOBEC3A.
JOURNAL Nat Biotechnol. 2018 Oct 1. pii: nbt.4261. doi: 10.1038/nbt.4261.
PUBMED 30272679
REFERENCE 2 (bases 1 to 17062)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..17062
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(919..1548)
/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 2849..2873
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 3061..3083
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 3075..3092
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 3076..3092
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 3805..4596
/codon_start=1
/gene="aadA"
/product="aminoglycoside adenylyltransferase (Murphy,
1985)"
/label=SmR
/note="confers resistance to spectinomycin and
streptomycin"
/translation="MGEAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
misc_RNA 4748..4823
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 5437..5781
/label=CaMV 35S promoter
/note="strong constitutive promoter from cauliflower mosaic
virus"
CDS 5930..5950
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5951..6634
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
WATGLK"
CDS 6686..10786
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 10787..10834
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 10856..11104
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 11117..11137
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(11793..11809)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11793..11809)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(11806..11828)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 11817..11833
/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(11841..11871)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11886..11907
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 12098..12776
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
CDS 12821..13372
/codon_start=1
/gene="Streptomyces hygroscopicus bar"
/product="phosphinothricin acetyltransferase"
/label=BlpR
/note="confers resistance to bialophos or phosphinothricin"
/translation="MSPERRPADIRRATEADMPAVCTIVNHYIETSTVNFRTEPQEPQE
WTDDLVRLRERYPWLVAEVDGEVAGIAYAGPWKARNAYDWTAESTVYVSPRHQRTGLGS
TLYTHLLKSLEAQGFKSVVAVIGLPNDPSVRMHEALGYAPRGMLRAAGFKHGNWHDVGF
WQLDFSLPVPPRPVLPVTEI"
polyA_signal 13379..13553
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 13631..13655
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 14080..14874
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 14961..15549
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 15450..15469
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 15703..15720
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 15735..15875
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(15801..15820)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 15961..15983
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 16219..16413
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(16479..490)
/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"
ORIGIN
1 cgggcttgtc tcccttccct tcccggtatc ggttcatgga ttcggttaga tgggaaaccg
61 ccatcagtac caggtcgtaa tcccacacac tggccatgcc ggccggccct gcggaaacct
121 ctacgtgccc gtctggaagc tcgtagcgga tcacctcgcc agctcgtcgg tcacgcttcg
181 acagacggaa aacggccacg tccatgatgc tgcgactatc gcgggtgccc acgtcataga
241 gcatcggaac gaaaaaatct ggttgctcgt cgcccttggg cggcttccta atcgacggcg
301 caccggctgc cggcggttgc cgggattctt tgcggattcg atcagcggcc gcttgccacg
361 attcaccggg gcgtgcttct gcctcgatgc gttgccgctg ggcggcctgc gcggccttca
421 acttctccac caggtcatca cccagcgccg cgccgatttg taccgggccg gatggtttgc
481 gaccgctcac gccgattcct cgggcttggg ggttccagtg ccattgcagg gccggcaggc
541 aacccagccg cttacgcctg gccaaccgcc cgttcctcca cacatggggc attccacggc
601 gtcggtgcct ggttgttctt gattttccat gccgcctcct ttagccgcta aaattcatct
661 actcatttat tcatttgctc atttactctg gtagctgcgc gatgtattca gatagcagct
721 cggtaatggt cttgccttgg cgtaccgcgt acatcttcag cttggtgtga tcctccgccg
781 gcaactgaaa gttgacccgc ttcatggctg gcgtgtctgc caggctggcc aacgttgcag
841 ccttgctgct gcgtgcgctc ggacggccgg cacttagcgt gtttgtgctt ttgctcattt
901 tctctttacc tcattaactc aaatgagttt tgatttaatt tcagcggcca gcgcctggac
961 ctcgcgggca gcgtcgccct cgggttctga ttcaagaacg gttgtgccgg cggcggcagt
1021 gcctgggtag ctcacgcgct gcgtgatacg ggactcaaga atgggcagct cgtacccggc
1081 cagcgcctcg gcaacctcac cgccgatgcg cgtgcctttg atcgcccgcg acacgacaaa
1141 ggccgcttgt agccttccat ccgtgacctc aatgcgctgc ttaaccagct ccaccaggtc
1201 ggcggtggcc catatgtcgt aagggcttgg ctgcaccgga atcagcacga agtcggctgc
1261 cttgatcgcg gacacagcca agtccgccgc ctggggcgct ccgtcgatca ctacgaagtc
1321 gcgccggccg atggccttca cgtcgcggtc aatcgtcggg cggtcgatgc cgacaacggt
1381 tagcggttga tcttcccgca cggccgccca atcgcgggca ctgccctggg gatcggaatc
1441 gactaacaga acatcggccc cggcgagttg cagggcgcgg gctagatggg ttgcgatggt
1501 cgtcttgcct gacccgcctt tctggttaag tacagcgata accttcatgc gttccccttg
1561 cgtatttgtt tatttactca tcgcatcata tacgcagcga ccgcatgacg caagctgttt
1621 tactcaaata cacatcacct ttttagacgg cggcgctcgg tttcttcagc ggccaagctg
1681 gccggccagg ccgccagctt ggcatcagac aaaccggcca ggatttcatg cagccgcacg
1741 gttgagacgt gcgcgggcgg ctcgaacacg tacccggccg cgatcatctc cgcctcgatc
1801 tcttcggtaa tgaaaaacgg ttcgtcctgg ccgtcctggt gcggtttcat gcttgttcct
1861 cttggcgttc attctcggcg gccgccaggg cgtcggcctc ggtcaatgcg tcctcacgga
1921 aggcaccgcg ccgcctggcc tcggtgggcg tcacttcctc gctgcgctca agtgcgcggt
1981 acagggtcga gcgatgcacg ccaagcagtg cagccgcctc tttcacggtg cggccttcct
2041 ggtcgatcag ctcgcgggcg tgcgcgatct gtgccggggt gagggtaggg cgggggccaa
2101 acttcacgcc tcgggccttg gcggcctcgc gcccgctccg ggtgcggtcg atgattaggg
2161 aacgctcgaa ctcggcaatg ccggcgaaca cggtcaacac catgcggccg gccggcgtgg
2221 tggtgtcggc ccacggctct gccaggctac gcaggcccgc gccggcctcc tggatgcgct
2281 cggcaatgtc cagtaggtcg cgggtgctgc gggccaggcg gtctagcctg gtcactgtca
2341 caacgtcgcc agggcgtagg tggtcaagca tcctggccag ctccgggcgg tcgcgcctgg
2401 tgccggtgat cttctcggaa aacagcttgg tgcagccggc cgcgtgcagt tcggcccgtt
2461 ggttggtcaa gtcctggtcg tcggtgctga cgcgggcata gcccagcagg ccagcggcgg
2521 cgctcttgtt catggcgtaa tgtctccggt tctagtcgca agtattctac tttatgcgac
2581 taaaacacgc gacaagaaaa cgccaggaaa agggcagggc ggcagcctgt cgcgtaactt
2641 aggacttgtg cgacatgtcg ttttcagaag acggctgcac tgaacgtcag aagccgactg
2701 cactatagca gcggaggggt tggatcaaag tactttgatc ccgaggggaa ccctgtggtt
2761 ggcatgcaca tacaaatgga cgaacggata aaccttttca cgccctttta aatatccgat
2821 tattctaata aacgctcttt tctcttaggt ttacccgcca atatatcctg tcaaacactg
2881 atagtttaaa ctgaaggcgg gaaacgacaa tctgatccaa gctcaagctg ctctagcatt
2941 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
3001 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
3061 cccagtcacg acgttgtaaa acgacggcca gtgccaagct tcgacttgcc ttccgcacaa
3121 tacatcattt cttcttagct ttttttcttc ttcttcgttc atacagtttt tttttgttta
3181 tcagcttaca ttttcttgaa ccgtagcttt cgttttcttc tttttaactt tccattcgga
3241 gtttttgtat cttgtttcat agtttgtccc aggattagaa tgattaggca tcgaaccttc
3301 aagaatttga ttgaataaaa catcttcatt cttaagatat gaagataatc ttcaaaaggc
3361 ccctgggaat ctgaaagaag agaagcaggc ccatttatat gggaaagaac aatagtattt
3421 cttatatagg cccatttaag ttgaaaacaa tcttcaaaag tcccacatcg cttagataag
3481 aaaacgaagc tgagtttata tacagctaga gtcgaagtag tgattgggag accaacccag
3541 tggacataag cctgttcggt tcgtaagctg taatgcaagt agcgtatgcg ctcacgcaac
3601 tggtccagaa ccttgaccga acgcagcggt ggtaacggcg cagtggcggt tttcatggct
3661 tgttatgact gtttttttgg ggtacagtct atgcctcggg catccaagca gcaagcgcgt
3721 tacgccgtgg gtcgatgttt gatgttatgg agcagcaacg atgttacgca gcagggcagt
3781 cgccctaaaa caaagttaaa catcatgggg gaagcggtga tcgccgaagt atcgactcaa
3841 ctatcagagg tagttggcgt catcgagcgc catctcgaac cgacgttgct ggccgtacat
3901 ttgtacggct ccgcagtgga tggcggcctg aagccacaca gtgatattga tttgctggtt
3961 acggtgaccg taaggcttga tgaaacaacg cggcgagctt tgatcaacga ccttttggaa
4021 acttcggctt cccctggaga gagcgagatt ctccgcgctg tagaagtcac cattgttgtg
4081 cacgacgaca tcattccgtg gcgttatcca gctaagcgcg aactgcaatt tggagaatgg
4141 cagcgcaatg acattcttgc aggtatcttc gagccagcca cgatcgacat tgatctggct
4201 atcttgctga caaaagcaag agaacatagc gttgccttgg taggtccagc ggcggaggaa
4261 ctctttgatc cggttcctga acaggatcta tttgaggcgc taaatgaaac cttaacgcta
4321 tggaactcgc cgcccgactg ggctggcgat gagcgaaatg tagtgcttac gttgtcccgc
4381 atttggtaca gcgcagtaac cggcaaaatc gcgccgaagg atgtcgctgc cgactgggca
4441 atggagcgcc tgccggccca gtatcagccc gtcatacttg aagctagaca ggcttatctt
4501 ggacaagaag aagatcgctt ggcctcgcgc gcagatcagt tggaagaatt tgtccactac
4561 gtgaaaggcg agatcaccaa ggtagtcggc aaataatgtc tagctagaaa ttcgttcaag
4621 ccgacgccgc ttcgcggcgc ggcttaactc aagcgttaga tgcactaagc acataattgc
4681 tcacagccaa actatcaggt caagtctgct tttattattt ttaagcgtgc ataataagcc
4741 ggtctcggtt ttagagctag aaatagcaag ttaaaataag gctagtccgt tatcaacttg
4801 aaaaagtggc accgagtcgg tgcttttttt tgcaaaattt tccagatcga tttcttcttc
4861 ctctgttctt cggcgttcaa tttctggggt tttctcttcg ttttctgtaa ctgaaaccta
4921 aaatttgacc taaaaaaaat ctcaaataat atgattcagt ggttttgtac ttttcagtta
4981 gttgagtttt gcagttccga tgagataaac caataagctt gcatgcctgc aggtcaacat
5041 ggtggagcac gacacacttg tctactccaa aaatatcaaa gatacagtct cagaagacca
5101 aagggcaatt gagacttttc aacaaagggt aatatccgga aacctcctcg gattccattg
5161 cccagctatc tgtcacttta ttgtgaagat agtggaaaag gaaggtggct cctacaaatg
5221 ccatcattgc gataaaggaa aggccatcgt tgaagatgcc tctgccgaca gtggtcccaa
5281 agatggaccc ccacccacga ggagcatcgt ggaaaaagaa gacgttccaa ccacgtcttc
5341 aaagcaagtg gattgatgtg ataacatggt ggagcacgac acacttgtct actccaaaaa
5401 tatcaaagat acagtctcag aagaccaaag ggcaattgag acttttcaac aaagggtaat
5461 atccggaaac ctcctcggat tccattgccc agctatctgt cactttattg tgaagatagt
5521 ggaaaaggaa ggtggctcct acaaatgcca tcattgcgat aaaggaaagg ccatcgttga
5581 agatgcctct gccgacagtg gtcccaaaga tggaccccca cccacgagga gcatcgtgga
5641 aaaagaagac gttccaacca cgtcttcaaa gcaagtggat tgatgtgata tctccactga
5701 cgtaagggat gacgcacaat cccactatcc ttcgcaagac ccttcctcta tataaggaag
5761 ttcatttcat ttggagagga cctcgacctc aacacaacat atacaaaaca aacgaatctc
5821 aagcaatcaa gcattctact tctattgcag caatttaaat catttctttt aaagcaaaag
5881 caattttctg aaaattttca ccatttacga acgatactcg agtaatatgc caaagaagaa
5941 gaggaaggtt tcatcggaga ccggccctgt tgctgttgac cccaccctgc ggcggagaat
6001 cgagccacac gagttcgagg tgttcttcga cccaagggag ctccgcaagg agacgtgcct
6061 cctgtacgag atcaactggg gcggcaggca ctccatctgg aggcacacca gccaaaacac
6121 caacaagcac gtggaggtca acttcatcga gaagttcacc accgagaggt acttctgccc
6181 aaacacccgc tgctccatca cctggttcct gtcctggagc ccatgcggcg agtgctccag
6241 ggccatcacc gagttcctca gccgctaccc acacgtcacc ctgttcatct acatcgccag
6301 gctctaccac cacgccgacc caaggaacag gcagggcctc cgcgacctga tctccagcgg
6361 cgtgaccatc caaatcatga ccgagcagga gtccggctac tgctggagga acttcgtcaa
6421 ctactcccca agcaacgagg cccactggcc aaggtaccca cacctctggg tgcgcctcta
6481 cgtgctcgag ctgtactgca tcatcctcgg cctgccacca tgcctcaaca tcctgaggcg
6541 caagcaacca cagctgacct tcttcaccat cgccctccaa agctgccact accagaggct
6601 cccaccacac atcctgtggg ctaccggcct caagtccggc agcgagacgc caggcacctc
6661 cgagagcgct acgcctgaac ttaaggacaa gaagtactcg atcggcctcg ccatcgggac
6721 gaactcagtt ggctgggccg tgatcaccga cgagtacaag gtgccctcta agaagttcaa
6781 ggtcctgggg aacaccgacc gccattccat caagaagaac ctcatcggcg ctctcctgtt
6841 cgacagcggg gagaccgctg aggctacgag gctcaagaga accgctaggc gccggtacac
6901 gagaaggaag aacaggatct gctacctcca agagattttc tccaacgaga tggccaaggt
6961 tgacgattca ttcttccacc gcctggagga gtctttcctc gtggaggagg ataagaagca
7021 cgagcggcat cccatcttcg gcaacatcgt ggacgaggtt gcctaccacg agaagtaccc
7081 tacgatctac catctgcgga agaagctcgt ggactccacc gataaggcgg acctcagact
7141 gatctacctc gctctggccc acatgatcaa gttccgcggc catttcctga tcgaggggga
7201 tctcaaccca gacaacagcg atgttgacaa gctgttcatc caactcgtgc agacctacaa
7261 ccaactcttc gaggagaacc cgatcaacgc ctctggcgtg gacgcgaagg ctatcctgtc
7321 cgcgaggctc tcgaagtcca ggaggctgga gaacctgatc gctcagctcc caggcgagaa
7381 gaagaacggc ctgttcggga acctcatcgc tctcagcctg gggctcaccc cgaacttcaa
7441 gtcgaacttc gatctcgctg aggacgccaa gctgcaactc tccaaggaca cctacgacga
7501 tgacctcgat aacctcctgg cccagatcgg cgatcaatac gcggacctgt tcctcgctgc
7561 caagaacctg tcggacgcca tcctcctgtc agatatcctc cgcgtgaaca ccgagatcac
7621 gaaggctcca ctctctgcct ccatgatcaa gcgctacgac gagcaccatc aggatctgac
7681 cctcctgaag gcgctggtcc gccaacagct cccggagaag tacaaggaga ttttcttcga
7741 tcagtcgaag aacggctacg ctgggtacat cgacggcggg gcctcacaag aggagttcta
7801 caagttcatc aagccaatcc tggagaagat ggacggcacg gaggagctcc tggtgaagct
7861 caacagggag gacctcctgc ggaagcagag aaccttcgat aacggcagca tcccccacca
7921 aatccatctc ggggagctgc acgccatcct gagaaggcaa gaggacttct accctttcct
7981 caaggataac cgggagaaga tcgagaagat cctgaccttc agaatcccat actacgtcgg
8041 ccctctcgcg cgggggaact caagattcgc ttggatgacc cgcaagtctg aggagaccat
8101 cacgccgtgg aacttcgagg aggtggtgga caagggcgct agcgctcagt cgttcatcga
8161 gaggatgacc aacttcgaca agaacctgcc caacgagaag gtgctcccta agcactcgct
8221 cctgtacgag tacttcaccg tctacaacga gctcacgaag gtgaagtacg tcaccgaggg
8281 catgcgcaag ccagcgttcc tgtccgggga gcagaagaag gctatcgtgg acctcctgtt
8341 caagaccaac cggaaggtca cggttaagca actcaaggag gactacttca agaagatcga
8401 gtgcttcgat tcggtcgaga tcagcggcgt tgaggaccgc ttcaacgcca gcctcgggac
8461 ctaccacgat ctcctgaaga tcatcaagga taaggacttc ctggacaacg aggagaacga
8521 ggatatcctg gaggacatcg tgctgaccct cacgctgttc gaggacaggg agatgatcga
8581 ggagcgcctg aagacgtacg cccatctctt cgatgacaag gtcatgaagc aactcaagcg
8641 ccggagatac accggctggg ggaggctgtc ccgcaagctc atcaacggca tccgggacaa
8701 gcagtccggg aagaccatcc tcgacttcct caagagcgat ggcttcgcca acaggaactt
8761 catgcaactg atccacgatg acagcctcac cttcaaggag gatatccaaa aggctcaagt
8821 gagcggccag ggggactcgc tgcacgagca tatcgcgaac ctcgctggct cccccgcgat
8881 caagaagggc atcctccaga ccgtgaaggt tgtggacgag ctcgtgaagg tcatgggccg
8941 gcacaagcct gagaacatcg tcatcgagat ggccagagag aaccaaacca cgcagaaggg
9001 gcaaaagaac tctagggagc gcatgaagcg catcgaggag ggcatcaagg agctggggtc
9061 ccaaatcctc aaggagcacc cagtggagaa cacccaactg cagaacgaga agctctacct
9121 gtactacctc cagaacggca gggatatgta cgtggaccaa gagctggata tcaaccgcct
9181 cagcgattac gacgtcgatc atatcgttcc ccagtctttc ctgaaggatg actccatcga
9241 caacaaggtc ctcaccaggt cggacaagaa ccgcggcaag tcagataacg ttccatctga
9301 ggaggtcgtt aagaagatga agaactactg gaggcagctc ctgaacgcca agctgatcac
9361 gcaaaggaag ttcgacaacc tcaccaaggc tgagagaggc gggctctcag agctggacaa
9421 ggccggcttc atcaagcggc agctggtcga gaccagacaa atcacgaagc acgttgcgca
9481 aatcctcgac tctcggatga acacgaagta cgatgagaac gacaagctga tcagggaggt
9541 taaggtgatc accctgaagt ctaagctcgt ctccgacttc aggaaggatt tccagttcta
9601 caaggttcgc gagatcaaca actaccacca tgcccatgac gcttacctca acgctgtggt
9661 cggcaccgct ctgatcaaga agtacccaaa gctggagtcc gagttcgtgt acggggacta
9721 caaggtttac gatgtgcgca agatgatcgc caagtcggag caagagatcg gcaaggctac
9781 cgccaagtac ttcttctact caaacatcat gaacttcttc aagaccgaga tcacgctggc
9841 caacggcgag atccggaaga gaccgctcat cgagaccaac ggcgagacgg gggagatcgt
9901 gtgggacaag ggcagggatt tcgcgaccgt ccgcaaggtt ctctccatgc cccaggtgaa
9961 catcgtcaag aagaccgagg tccaaacggg cgggttctca aaggagtcta tcctgcctaa
10021 gcggaacagc gacaagctca tcgccagaaa gaaggactgg gacccaaaga agtacggcgg
10081 gttcgacagc cctaccgtgg cctactcggt cctggttgtg gcgaaggttg agaagggcaa
10141 gtccaagaag ctcaagagcg tgaaggagct cctggggatc accatcatgg agaggtccag
10201 cttcgagaag aacccaatcg acttcctgga ggccaagggc tacaaggagg tgaagaagga
10261 cctgatcatc aagctcccga agtactctct cttcgagctg gagaacggca ggaagagaat
10321 gctggcttcc gctggcgagc tccagaaggg gaacgagctc gcgctgccaa gcaagtacgt
10381 gaacttcctc tacctggctt cccactacga gaagctcaag ggcagcccgg aggacaacga
10441 gcaaaagcag ctgttcgtcg agcagcacaa gcattacctc gacgagatca tcgagcaaat
10501 ctccgagttc agcaagcgcg tgatcctcgc cgacgcgaac ctggataagg tcctctccgc
10561 ctacaacaag caccgggaca agcccatcag agagcaagcg gagaacatca tccatctctt
10621 caccctgacg aacctcggcg ctcctgctgc tttcaagtac ttcgacacca cgatcgatcg
10681 gaagagatac acctccacga aggaggtcct ggacgcgacc ctcatccacc agtcgatcac
10741 cggcctgtac gagacgagga tcgacctctc acaactcggc ggggataaga gacccgcagc
10801 aaccaagaag gcagggcaag caaagaagaa gaagacgcgt gactccggcg gcagcaccaa
10861 cctgtccgac atcatcgaga aggagacggg caagcaactc gtgatccagg agagcatcct
10921 catgctgcca gaggaggtgg aggaggtcat cggcaacaag ccagagtccg acatcctggt
10981 gcacaccgcc tacgacgagt ccaccgacga gaacgtcatg ctcctgacca gcgacgcccc
11041 agagtacaag ccatgggccc tcgtcatcca ggacagcaac ggggagaaca agatcaagat
11101 gctgtcgggg gggagcccaa agaagaagcg gaaggtgtag ctcagagctt tcgttcgtat
11161 catcggtttc gacaacgttc gtcaagttca atgcatcagt ttcattgcgc acacaccaga
11221 atcctactga gtttgagtat tatggcattg ggaaaactgt ttttcttgta ccatttgttg
11281 tgcttgtaat ttactgtgtt ttttattcgg ttttcgctat cgaactgtga aatggaaatg
11341 gatggagaag agttaatgaa tgatatggtc cttttgttca ttctcaaatt aatattattt
11401 gttttttctc ttatttgttg tgtgttgaat ttgaaattat aagagatatg caaacatttt
11461 gttttgagta aaaatgtgtc aaatcgtggc ctctaatgac cgaagttaat atgaggagta
11521 aaacacttgt agttgtacca ttatgcttat tcactaggca acaaatatat tttcagacct
11581 agaaaagctg caaatgttac tgaatacaag tatgtcctct tgtgttttag acatttatga
11641 actttccttt atgtaatttt ccagaatcct tgtcagattc taatcattgc tttataatta
11701 tagttatact catggatttg tagttgagta tgaaaatatt ttttaatgca ttttatgact
11761 tgccaattga ttgacaacga attcgtaatc atgtcatagc tgtttcctgt gtgaaattgt
11821 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
11881 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
11941 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
12001 cgtattggct agagcagctt gccaacatgg tggagcacga cactctcgtc tactccaaga
12061 atatcaaaga tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa
12121 tatcgggaaa cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag
12181 tagaaaagga aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc
12241 aagatgcctc tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg
12301 aaaaagaaga cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat aacatggtgg
12361 agcacgacac tctcgtctac tccaagaata tcaaagatac agtctcagaa gaccaaaggg
12421 ctattgagac ttttcaacaa agggtaatat cgggaaacct cctcggattc cattgcccag
12481 ctatctgtca cttcatcaaa aggacagtag aaaaggaagg tggcacctac aaatgccatc
12541 attgcgataa aggaaaggct atcgttcaag atgcctctgc cgacagtggt cccaaagatg
12601 gacccccacc cacgaggagc atcgtggaaa aagaagacgt tccaaccacg tcttcaaagc
12661 aagtggattg atgtgatatc tccactgacg taagggatga cgcacaatcc cactatcctt
12721 cgcaagaccc ttcctctata taaggaagtt catttcattt ggagaggaca cgctgaaatc
12781 accagtctct ctctacaaat ctatctctct cgagtctacc atgagcccag aacgacgccc
12841 ggccgacatc cgccgtgcca ccgaggcgga catgccggcg gtctgcacca tcgtcaacca
12901 ctacatcgag acaagcacgg tcaacttccg taccgagccg caggaaccgc aggagtggac
12961 ggacgacctc gtccgtctgc gggagcgcta tccctggctc gtcgccgagg tggacggcga
13021 ggtcgccggc atcgcctacg cgggcccctg gaaggcacgc aacgcctacg actggacggc
13081 cgagtcgacc gtgtacgtct ccccccgcca ccagcggacg ggactgggct ccacgctcta
13141 cacccacctg ctgaagtccc tggaggcaca gggcttcaag agcgtggtcg ctgtcatcgg
13201 gctgcccaac gacccgagcg tgcgcatgca cgaggcgctc ggatatgccc cccgcggcat
13261 gctgcgggcg gccggcttca agcacgggaa ctggcatgac gtgggtttct ggcagctgga
13321 cttcagcctg ccggtaccgc cccgtccggt cctgcccgtc accgagattt gactcgagtt
13381 tctccataat aatgtgtgag tagttcccag ataagggaat tagggttcct atagggtttc
13441 gctcatgtgt tgagcatata agaaaccctt agtatgtatt tgtatttgta aaatacttct
13501 atcaataaaa tttctaattc ctaaaaccaa aatccagtac taaaatccag atcccccgaa
13561 ttaattcggc gttaattcag tacattaaaa acgtccgcaa tgtgttatta agttgtctaa
13621 gcgtcaattt gtttacacca caatatatcc tgccaccagc cagccaacag ctccccgacc
13681 ggcagctcgg cacaaaatca ccactcgata caggcagccc atcagtccgg gacggcgtca
13741 gcgggagagc cgttgtaagg cggcagactt tgctcatgtt accgatgcta ttcggaagaa
13801 cggcaactaa gctgccgggt ttgaaacacg gatgatctcg cggagggtag catgttgatt
13861 gtaacgatga cagagcgttg ctgcctgtga tcaccgcggt ttcaaaatcg gctccgtcga
13921 tactatgtta tacgccaact ttgaaaacaa ctttgaaaaa gctgttttct ggtatttaag
13981 gttttagaat gcaaggaaca gtgaattgga gttcgtcttg ttataattag cttcttgggg
14041 tatctttaaa tactgtagaa aagaggaagg aaataataaa tggctaaaat gagaatatca
14101 ccggaattga aaaaactgat cgaaaaatac cgctgcgtaa aagatacgga aggaatgtct
14161 cctgctaagg tatataagct ggtgggagaa aatgaaaacc tatatttaaa aatgacggac
14221 agccggtata aagggaccac ctatgatgtg gaacgggaaa aggacatgat gctatggctg
14281 gaaggaaagc tgcctgttcc aaaggtcctg cactttgaac ggcatgatgg ctggagcaat
14341 ctgctcatga gtgaggccga tggcgtcctt tgctcggaag agtatgaaga tgaacaaagc
14401 cctgaaaaga ttatcgagct gtatgcggag tgcatcaggc tctttcactc catcgacata
14461 tcggattgtc cctatacgaa tagcttagac agccgcttag ccgaattgga ttacttactg
14521 aataacgatc tggccgatgt ggattgcgaa aactgggaag aagacactcc atttaaagat
14581 ccgcgcgagc tgtatgattt tttaaagacg gaaaagcccg aagaggaact tgtcttttcc
14641 cacggcgacc tgggagacag caacatcttt gtgaaagatg gcaaagtaag tggctttatt
14701 gatcttggga gaagcggcag ggcggacaag tggtatgaca ttgccttctg cgtccggtcg
14761 atcagggagg atatcgggga agaacagtat gtcgagctat tttttgactt actggggatc
14821 aagcctgatt gggagaaaat aaaatattat attttactgg atgaattgtt ttagtaccta
14881 gaatgcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
14941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
15001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
15061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
15121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
15181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
15241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
15301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
15361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
15421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
15481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
15541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
15601 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
15661 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
15721 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
15781 cgcatatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtataca
15841 ctccgctatc gctacgtgac tgggtcatgg ctgcgccccg acacccgcca acacccgctg
15901 acgcgccctg acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct
15961 ccgggagctg catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg aggcagggtg
16021 ccttgatgtg ggcgccggcg gtcgagtggc gacggcgcgg cttgtccgcg ccctggtaga
16081 ttgcctggcc gtaggccagc catttttgag cggccagcgg ccgcgatagg ccgacgcgaa
16141 gcggcggggc gtagggagcg cagcgaccga agggtaggcg ctttttgcag ctcttcggct
16201 gtgcgctggc cagacagtta tgcacaggcc aggcgggttt taagagtttt aataagtttt
16261 aaagagtttt aggcggaaaa atcgcctttt ttctctttta tatcagtcac ttacatgtgt
16321 gaccggttcc caatgtacgg ctttgggttc ccaatgtacg ggttccggtt cccaatgtac
16381 ggctttgggt tcccaatgta cgtgctatcc acaggaaaca gaccttttcg acctttttcc
16441 cctgctaggg caatttgccc tagcatctgc tccgtacatt aggaaccggc ggatgcttcg
16501 ccctcgatca ggttgcggta gcgcatgact aggatcgggc cagcctgccc cgcctcctcc
16561 ttcaaatcgt actccggcag gtcatttgac ccgatcagct tgcgcacggt gaaacagaac
16621 ttcttgaact ctccggcgct gccactgcgt tcgtagatcg tcttgaacaa ccatctggct
16681 tctgccttgc ctgcggcgcg gcgtgccagg cggtagagaa aacggccgat gccgggatcg
16741 atcaaaaagt aatcggggtg aaccgtcagc acgtccgggt tcttgccttc tgtgatctcg
16801 cggtacatcc aatcagctag ctcgatctcg atgtactccg gccgcccggt ttcgctcttt
16861 acgatcttgt agcggctaat caaggcttca ccctcggata ccgtcaccag gcggccgttc
16921 ttggccttct tcgtacgctg catggcaacg tgcgtggtgt ttaaccgaat gcaggtttct
16981 accaggtcgt ctttctgctt tccgccatcg gctcgccggc agaacttgag tacgtccgca
17041 acgtgtggac ggaacacgcg gc
//
