| 质粒名称: | pH-ABE8e |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 200774 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | pHUE411 (modified from pCambia) |
| 完整质粒大小: | 16936bp |
| 载体类型: | Plant Expression, CRISPR |
| 筛选标记: | Hygromycin |
| 载体抗性: | Kanamycin |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Cas9(D10A)-TadA8e / TadA8e-32aa linker-nCas9(D10A)-3NLS |
| 物种来源: | Streptococcus pyogenes for Cas9; E. coli for TadA |
| 基因突变: | D10A for Cas9; W23R, H36L, P48A, R51L, L84F, A106V, D108N, A109S, T111R, D119N, H122N, H123Y, S146C, F149Y, R152P, E155V, I156F, K157N, T166I, D167N for TadA |
| 启动子: | maize Ubiquitin-1, OsU3 |
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | n/a |
|
文献引用方法 |
|
| 材料和方法部分: | pH-ABE8e was a gift from Caixia Gao (Addgene plasmid # 200774 ; http://n2t.net/addgene:200774 ; RRID:Addgene_200774) |
| 参考文献部分: | Tuning plant phenotypes by precise, graded downregulation of gene expression. Xue C, Qiu F, Wang Y, Li B, Zhao KT, Chen K, Gao C. Nat Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w. 10.1038/s41587-023-01707-w PubMed 36894598 |
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 pH-ABE8e 16936 bp ds-DNA circular SYN 05-JUL-2023
DEFINITION For plant adenine base editing in rice plants or monocotyledons
protoplasts.
ACCESSION .
VERSION .
KEYWORDS pH-ABE8e
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16936)
AUTHORS Xue C, Qiu F, Wang Y, Li B, Zhao KT, Chen K, Gao C
TITLE Tuning plant phenotypes by precise, graded downregulation of gene
expression.
JOURNAL Nat Biotechnol. 2023 Mar 9. doi: 10.1038/s41587-023-01707-w.
PUBMED 36894598
REFERENCE 2 (bases 1 to 16936)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jul 5, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..16936
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(578..1207)
/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 2508..2532
/label=RB T-DNA repeat
/note="right border repeat from nopaline C58 T-DNA"
primer_bind 2720..2742
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 2734..2751
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 2735..2751
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 2818..3197
/label=OsU3 promoter
/note="Oryza sativa (rice) snRNA U3 promoter"
terminator 3305..3324
/label=SUP4 terminator
/note="transcription terminator for the S. cerevisiae SUP4
tRNA gene"
promoter 3337..5329
/label=Ubi promoter
/note="maize polyubiquitin gene promoter"
CDS 5940..10040
/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"
RBS 9953..9961
/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 10053..10073
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10086..10106
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 10119..10139
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(10798..10814)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10798..10814)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10811..10833)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10822..10838
/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(10846..10876)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10891..10912
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11103..11779
/label=CaMV 35S promoter (enhanced)
/note="cauliflower mosaic virus 35S promoter with a
duplicated enhancer region"
primer_bind 11716..11736
/label=35S promoter
/note="CaMV 35S promoter, forward primer"
CDS 11846..12871
/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 12912..13086
/label=CaMV poly(A) signal
/note="cauliflower mosaic virus polyadenylation signal"
misc_feature 13164..13188
/label=LB T-DNA repeat
/note="left border repeat from nopaline C58 T-DNA"
CDS 13613..14407
/codon_start=1
/gene="aphA-3"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
FCVRSIREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
rep_origin 14494..15082
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 14983..15002
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 15236..15253
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 15268..15408
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(15334..15353)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 15494..15516
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
rep_origin 15752..15946
/label=pVS1 oriV
/note="origin of replication for the Pseudomonas plasmid
pVS1 (Heeb et al., 2000)"
CDS complement(16012..149)
/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 tcagcggccg cttgccacga ttcaccgggg cgtgcttctg cctcgatgcg ttgccgctgg
61 gcggcctgcg cggccttcaa cttctccacc aggtcatcac ccagcgccgc gccgatttgt
121 accgggccgg atggtttgcg accgctcacg ccgattcctc gggcttgggg gttccagtgc
181 cattgcaggg ccggcaggca acccagccgc ttacgcctgg ccaaccgccc gttcctccac
241 acatggggca ttccacggcg tcggtgcctg gttgttcttg attttccatg ccgcctcctt
301 tagccgctaa aattcatcta ctcatttatt catttgctca tttactctgg tagctgcgcg
361 atgtattcag atagcagctc ggtaatggtc ttgccttggc gtaccgcgta catcttcagc
421 ttggtgtgat cctccgccgg caactgaaag ttgacccgct tcatggctgg cgtgtctgcc
481 aggctggcca acgttgcagc cttgctgctg cgtgcgctcg gacggccggc acttagcgtg
541 tttgtgcttt tgctcatttt ctctttacct cattaactca aatgagtttt gatttaattt
601 cagcggccag cgcctggacc tcgcgggcag cgtcgccctc gggttctgat tcaagaacgg
661 ttgtgccggc ggcggcagtg cctgggtagc tcacgcgctg cgtgatacgg gactcaagaa
721 tgggcagctc gtacccggcc agcgcctcgg caacctcacc gccgatgcgc gtgcctttga
781 tcgcccgcga cacgacaaag gccgcttgta gccttccatc cgtgacctca atgcgctgct
841 taaccagctc caccaggtcg gcggtggccc atatgtcgta agggcttggc tgcaccggaa
901 tcagcacgaa gtcggctgcc ttgatcgcgg acacagccaa gtccgccgcc tggggcgctc
961 cgtcgatcac tacgaagtcg cgccggccga tggccttcac gtcgcggtca atcgtcgggc
1021 ggtcgatgcc gacaacggtt agcggttgat cttcccgcac ggccgcccaa tcgcgggcac
1081 tgccctgggg atcggaatcg actaacagaa catcggcccc ggcgagttgc agggcgcggg
1141 ctagatgggt tgcgatggtc gtcttgcctg acccgccttt ctggttaagt acagcgataa
1201 ccttcatgcg ttccccttgc gtatttgttt atttactcat cgcatcatat acgcagcgac
1261 cgcatgacgc aagctgtttt actcaaatac acatcacctt tttagacggc ggcgctcggt
1321 ttcttcagcg gccaagctgg ccggccaggc cgccagcttg gcatcagaca aaccggccag
1381 gatttcatgc agccgcacgg ttgagacgtg cgcgggcggc tcgaacacgt acccggccgc
1441 gatcatctcc gcctcgatct cttcggtaat gaaaaacggt tcgtcctggc cgtcctggtg
1501 cggtttcatg cttgttcctc ttggcgttca ttctcggcgg ccgccagggc gtcggcctcg
1561 gtcaatgcgt cctcacggaa ggcaccgcgc cgcctggcct cggtgggcgt cacttcctcg
1621 ctgcgctcaa gtgcgcggta cagggtcgag cgatgcacgc caagcagtgc agccgcctct
1681 ttcacggtgc ggccttcctg gtcgatcagc tcgcgggcgt gcgcgatctg tgccggggtg
1741 agggtagggc gggggccaaa cttcacgcct cgggccttgg cggcctcgcg cccgctccgg
1801 gtgcggtcga tgattaggga acgctcgaac tcggcaatgc cggcgaacac ggtcaacacc
1861 atgcggccgg ccggcgtggt ggtgtcggcc cacggctctg ccaggctacg caggcccgcg
1921 ccggcctcct ggatgcgctc ggcaatgtcc agtaggtcgc gggtgctgcg ggccaggcgg
1981 tctagcctgg tcactgtcac aacgtcgcca gggcgtaggt ggtcaagcat cctggccagc
2041 tccgggcggt cgcgcctggt gccggtgatc ttctcggaaa acagcttggt gcagccggcc
2101 gcgtgcagtt cggcccgttg gttggtcaag tcctggtcgt cggtgctgac gcgggcatag
2161 cccagcaggc cagcggcggc gctcttgttc atggcgtaat gtctccggtt ctagtcgcaa
2221 gtattctact ttatgcgact aaaacacgcg acaagaaaac gccaggaaaa gggcagggcg
2281 gcagcctgtc gcgtaactta ggacttgtgc gacatgtcgt tttcagaaga cggctgcact
2341 gaacgtcaga agccgactgc actatagcag cggaggggtt ggatcaaagt actttgatcc
2401 cgaggggaac cctgtggttg gcatgcacat acaaatggac gaacggataa accttttcac
2461 gcccttttaa atatccgatt attctaataa acgctctttt ctcttaggtt tacccgccaa
2521 tatatcctgt caaacactga tagtttaaac tgaaggcggg aaacgacaat ctgatccaag
2581 ctcaagctgc tctagcattc gccattcagg ctgcgcaact gttgggaagg gcgatcggtg
2641 cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag gcgattaagt
2701 tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag tgccaagctt
2761 agtaattcat ccaggtcacc aagttctagg attttcagaa ctgcaactta ttttatcaag
2821 gaatctttaa acatacgaac agatcactta aagttcttct gaagcaactt aaagttatca
2881 ggcatgcatg gatcttggag gaatcagatg tgcagtcagg gaccatagca caagacaggc
2941 gtcttctact ggtgctacca gcaaatgctg gaagccggga acactgggta cgttggaaac
3001 cacgtgatgt gaagaagtaa gataaactgt aggagaaaag catttcgtag tgggccatga
3061 agcctttcag gacatgtatt gcagtatggg ccggcccatt acgcaattgg acgacaacaa
3121 agactagtat tagtaccacc tcggctatcc acatagatca aagctgattt aaaagagttg
3181 tgcagatgat ccgtggcgtg agaccctgca gggtctctgt ttaagagcta tgctggaaac
3241 agcatagcaa gtttaaataa ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg
3301 gtgctttttt tgttttttat gtctaagctt gcatgcctgc agtgcagcgt gacccggtcg
3361 tgcccctctc tagagataat gagcattgca tgtctaagtt ataaaaaatt accacatatt
3421 ttttttgtca cacttgtttg aagtgcagtt tatctatctt tatacatata tttaaacttt
3481 actctacgaa taatataatc tatagtacta caataatatc agtgttttag agaatcatat
3541 aaatgaacag ttagacatgg tctaaaggac aattgagtat tttgacaaca ggactctaca
3601 gttttatctt tttagtgtgc atgtgttctc cttttttttt gcaaatagct tcacctatat
3661 aatacttcat ccattttatt agtacatcca tttagggttt agggttaatg gtttttatag
3721 actaattttt ttagtacatc tattttattc tattttagcc tctaaattaa gaaaactaaa
3781 actctatttt agttttttta tttaataatt tagatataaa atagaataaa ataaagtgac
3841 taaaaattaa acaaataccc tttaagaaat taaaaaaact aaggaaacat ttttcttgtt
3901 tcgagtagat aatgccagcc tgttaaacgc cgtcgacgag tctaacggac accaaccagc
3961 gaaccagcag cgtcgcgtcg ggccaagcga agcagacggc acggcatctc tgtcgctgcc
4021 tctggacccc tctcgagagt tccgctccac cgttggactt gctccgctgt cggcatccag
4081 aaattgcgtg gcggagcggc agacgtgagc cggcacggca ggcggcctcc tcctcctctc
4141 acggcaccgg cagctacggg ggattccttt cccaccgctc cttcgctttc ccttcctcgc
4201 ccgccgtaat aaatagacac cccctccaca ccctctttcc ccaacctcgt gttgttcgga
4261 gcgcacacac acacaaccag atctccccca aatccacccg tcggcacctc cgcttcaagg
4321 tacgccgctc gtcctccccc cccccccctc tctaccttct ctagatcggc gttccggtcc
4381 atggttaggg cccggtagtt ctacttctgt tcatgtttgt gttagatccg tgtttgtgtt
4441 agatccgtgc tgctagcgtt cgtacacgga tgcgacctgt acgtcagaca cgttctgatt
4501 gctaacttgc cagtgtttct ctttggggaa tcctgggatg gctctagccg ttccgcagac
4561 gggatcgatt tcatgatttt ttttgtttcg ttgcataggg tttggtttgc ccttttcctt
4621 tatttcaata tatgccgtgc acttgtttgt cgggtcatct tttcatgctt ttttttgtct
4681 tggttgtgat gatgtggtct ggttgggcgg tcgttctaga tcggagtaga attctgtttc
4741 aaactacctg gtggatttat taattttgga tctgtatgtg tgtgccatac atattcatag
4801 ttacgaattg aagatgatgg atggaaatat cgatctagga taggtataca tgttgatgcg
4861 ggttttactg atgcatatac agagatgctt tttgttcgct tggttgtgat gatgtggtgt
4921 ggttgggcgg tcgttcattc gttctagatc ggagtagaat actgtttcaa actacctggt
4981 gtatttatta attttggaac tgtatgtgtg tgtcatacat cttcatagtt acgagtttaa
5041 gatggatgga aatatcgatc taggataggt atacatgttg atgtgggttt tactgatgca
5101 tatacatgat ggcatatgca gcatctattc atatgctcta accttgagta cctatctatt
5161 ataataaaca agtatgtttt ataattattt tgatcttgat atacttggat gatggcatat
5221 gcagcagcta tatgtggatt tttttagccc tgccttcata cgctatttat ttgcttggta
5281 ctgtttcttt tgtcgatgct caccctgttg tttggtgtta cttctgcagc cctaggatga
5341 gcgaagttga atttagccat gaatattgga tgcgccatgc acttacactt gcaaagagag
5401 cccgcgatga acgcgaagtt cctgttggag cagttcttgt tcttaacaac cgcgttatcg
5461 gagaaggatg gaaccgcgca atcggacttc atgatcctac agcacatgca gaaatcatgg
5521 cacttcgcca gggaggactt gttatgcaga actatcgcct tatcgatgca acactttatg
5581 ttacatttga accttgcgtt atgtgcgcag gagcaatgat ccatagccgc atcggacgcg
5641 ttgtgttcgg agtacgtaac agcaaacgcg gagcagcagg aagccttatg aacgttctta
5701 actatcctgg aatgaaccat cgcgttgaaa tcacagaagg aatccttgca gatgaatgcg
5761 cagcacttct ttgcgatttc taccgtatgc ctcgccaggt gttcaatgcg cagaagaagg
5821 cccaaagcag catcaactca ggtggtagct ccggcggctc gtcaggttcc gaaacacctg
5881 gaacaagcga aagcgcaaca cctgaatcta gtggaggaag ttctggtggg tcccttaagg
5941 acaagaagta ctcgatcggc ctcgccattg ggactaactc tgttggctgg gccgtgatca
6001 ccgacgagta caaggtgccc tcaaagaagt tcaaggtcct gggcaacacc gatcggcatt
6061 ccatcaagaa gaatctcatt ggcgctctcc tgttcgacag cggcgagacg gctgaggcta
6121 cgcggctcaa gcgcaccgcc cgcaggcggt acacgcgcag gaagaatcgc atctgctacc
6181 tgcaggagat tttctccaac gagatggcga aggttgacga ttctttcttc cacaggctgg
6241 aggagtcatt cctcgtggag gaggataaga agcacgagcg gcatccaatc ttcggcaaca
6301 ttgtcgacga ggttgcctac cacgagaagt accctacgat ctaccatctg cggaagaagc
6361 tcgtggactc cacagataag gcggacctcc gcctgatcta cctcgctctg gcccacatga
6421 ttaagttcag gggccatttc ctgatcgagg gggatctcaa cccggacaat agcgatgttg
6481 acaagctgtt catccagctc gtgcagacgt acaaccagct cttcgaggag aaccccatta
6541 atgcgtcagg cgtcgacgcg aaggctatcc tgtccgctag gctctcgaag tctcggcgcc
6601 tcgagaacct gatcgcccag ctgccgggcg agaagaagaa cggcctgttc gggaatctca
6661 ttgcgctcag cctggggctc acgcccaact tcaagtcgaa tttcgatctc gctgaggacg
6721 ccaagctgca gctctccaag gacacatacg acgatgacct ggataacctc ctggcccaga
6781 tcggcgatca gtacgcggac ctgttcctcg ctgccaagaa tctgtcggac gccatcctcc
6841 tgtctgatat tctcagggtg aacaccgaga ttacgaaggc tccgctctca gcctccatga
6901 tcaagcgcta cgacgagcac catcaggatc tgaccctcct gaaggcgctg gtcaggcagc
6961 agctccccga gaagtacaag gagatcttct tcgatcagtc gaagaacggc tacgctgggt
7021 acattgacgg cggggcctct caggaggagt tctacaagtt catcaagccg attctggaga
7081 agatggacgg cacggaggag ctgctggtga agctcaatcg cgaggacctc ctgaggaagc
7141 agcggacatt cgataacggc agcatcccac accagattca tctcggggag ctgcacgcta
7201 tcctgaggag gcaggaggac ttctaccctt tcctcaagga taaccgcgag aagatcgaga
7261 agattctgac tttcaggatc ccgtactacg tcggcccact cgctaggggc aactcccgct
7321 tcgcttggat gacccgcaag tcagaggaga cgatcacgcc gtggaacttc gaggaggtgg
7381 tcgacaaggg cgctagcgct cagtcgttca tcgagaggat gacgaatttc gacaagaacc
7441 tgccaaatga gaaggtgctc cctaagcact cgctcctgta cgagtacttc acagtctaca
7501 acgagctgac taaggtgaag tatgtgaccg agggcatgag gaagccggct ttcctgtctg
7561 gggagcagaa gaaggccatc gtggacctcc tgttcaagac caaccggaag gtcacggtta
7621 agcagctcaa ggaggactac ttcaagaaga ttgagtgctt cgattcggtc gagatctctg
7681 gcgttgagga ccgcttcaac gcctccctgg ggacctacca cgatctcctg aagatcatta
7741 aggataagga cttcctggac aacgaggaga atgaggatat cctcgaggac attgtgctga
7801 cactcactct gttcgaggac cgggagatga tcgaggagcg cctgaagact tacgcccatc
7861 tcttcgatga caaggtcatg aagcagctca agaggaggag gtacaccggc tgggggaggc
7921 tgagcaggaa gctcatcaac ggcattcggg acaagcagtc cgggaagacg atcctcgact
7981 tcctgaagag cgatggcttc gcgaaccgca atttcatgca gctgattcac gatgacagcc
8041 tcacattcaa ggaggatatc cagaaggctc aggtgagcgg ccagggggac tcgctgcacg
8101 agcatatcgc gaacctcgct ggctcgccag ctatcaagaa ggggattctg cagaccgtga
8161 aggttgtgga cgagctggtg aaggtcatgg gcaggcacaa gcctgagaac atcgtcattg
8221 agatggcccg ggagaatcag accacgcaga agggccagaa gaactcacgc gagaggatga
8281 agaggatcga ggagggcatt aaggagctgg ggtcccagat cctcaaggag cacccggtgg
8341 agaacacgca gctgcagaat gagaagctct acctgtacta cctccagaat ggccgcgata
8401 tgtatgtgga ccaggagctg gatattaaca ggctcagcga ttacgacgtc gatcatatcg
8461 ttccacagtc attcctgaag gatgactcca ttgacaacaa ggtcctcacc aggtcggaca
8521 agaaccgggg caagtctgat aatgttcctt cagaggaggt cgttaagaag atgaagaact
8581 actggcgcca gctcctgaat gccaagctga tcacgcagcg gaagttcgat aacctcacaa
8641 aggctgagag gggcgggctc tctgagctgg acaaggcggg cttcatcaag aggcagctgg
8701 tcgagacacg gcagatcact aagcacgttg cgcagattct cgactcacgg atgaacacta
8761 agtacgatga gaatgacaag ctgatccgcg aggtgaaggt catcaccctg aagtcaaagc
8821 tcgtctccga cttcaggaag gatttccagt tctacaaggt tcgggagatc aacaattacc
8881 accatgccca tgacgcgtac ctgaacgcgg tggtcggcac agctctgatc aagaagtacc
8941 caaagctcga gagcgagttc gtgtacgggg actacaaggt ttacgatgtg aggaagatga
9001 tcgccaagtc ggagcaggag attggcaagg ctaccgccaa gtacttcttc tactctaaca
9061 ttatgaattt cttcaagaca gagatcactc tggccaatgg cgagatccgg aagcgccccc
9121 tcatcgagac gaacggcgag acgggggaga tcgtgtggga caagggcagg gatttcgcga
9181 ccgtcaggaa ggttctctcc atgccacaag tgaatatcgt caagaagaca gaggtccaga
9241 ctggcgggtt ctctaaggag tcaattctgc ctaagcggaa cagcgacaag ctcatcgccc
9301 gcaagaagga ctgggatccg aagaagtacg gcgggttcga cagccccact gtggcctact
9361 cggtcctggt tgtggcgaag gttgagaagg gcaagtccaa gaagctcaag agcgtgaagg
9421 agctgctggg gatcacgatt atggagcgct ccagcttcga gaagaacccg atcgatttcc
9481 tggaggcgaa gggctacaag gaggtgaaga aggacctgat cattaagctc cccaagtact
9541 cactcttcga gctggagaac ggcaggaagc ggatgctggc ttccgctggc gagctgcaga
9601 aggggaacga gctggctctg ccgtccaagt atgtgaactt cctctacctg gcctcccact
9661 acgagaagct caagggcagc cccgaggaca acgagcagaa gcagctgttc gtcgagcagc
9721 acaagcatta cctcgacgag atcattgagc agatttccga gttctccaag cgcgtgatcc
9781 tggccgacgc gaatctggat aaggtcctct ccgcgtacaa caagcaccgc gacaagccaa
9841 tcagggagca ggctgagaat atcattcatc tcttcaccct gacgaacctc ggcgcccctg
9901 ctgctttcaa gtacttcgac acaactatcg atcgcaagag gtacacaagc actaaggagg
9961 tcctggacgc gaccctcatc caccagtcga ttaccggcct ctacgagacg cgcatcgacc
10021 tgtctcagct cgggggcgac tccggcggca gcccaaagaa gaagcggaag gtgtctggag
10081 gttctcctaa gaaaaagaga aaagtgtccg gcggctcccc gaagaagaag cgcaaggtgt
10141 aggagctcag agctttcgtt cgtatcatcg gtttcgacaa cgttcgtcaa gttcaatgca
10201 tcagtttcat tgcgcacaca ccagaatcct actgagtttg agtattatgg cattgggaaa
10261 actgtttttc ttgtaccatt tgttgtgctt gtaatttact gtgtttttta ttcggttttc
10321 gctatcgaac tgtgaaatgg aaatggatgg agaagagtta atgaatgata tggtcctttt
10381 gttcattctc aaattaatat tatttgtttt ttctcttatt tgttgtgtgt tgaatttgaa
10441 attataagag atatgcaaac attttgtttt gagtaaaaat gtgtcaaatc gtggcctcta
10501 atgaccgaag ttaatatgag gagtaaaaca cttgtagttg taccattatg cttattcact
10561 aggcaacaaa tatattttca gacctagaaa agctgcaaat gttactgaat acaagtatgt
10621 cctcttgtgt tttagacatt tatgaacttt cctttatgta attttccaga atccttgtca
10681 gattctaatc attgctttat aattatagtt atactcatgg atttgtagtt gagtatgaaa
10741 atatttttta atgcatttta tgacttgcca attgattgac aacgaattcg taatcatgtc
10801 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
10861 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
10921 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
10981 ccaacgcgcg gggagaggcg gtttgcgtat tggctagagc agcttgccaa catggtggag
11041 cacgacactc tcgtctactc caagaatatc aaagatacag tctcagaaga ccaaagggct
11101 attgagactt ttcaacaaag ggtaatatcg ggaaacctcc tcggattcca ttgcccagct
11161 atctgtcact tcatcaaaag gacagtagaa aaggaaggtg gcacctacaa atgccatcat
11221 tgcgataaag gaaaggctat cgttcaagat gcctctgccg acagtggtcc caaagatgga
11281 cccccaccca cgaggagcat cgtggaaaaa gaagacgttc caaccacgtc ttcaaagcaa
11341 gtggattgat gtgaacatgg tggagcacga cactctcgtc tactccaaga atatcaaaga
11401 tacagtctca gaagaccaaa gggctattga gacttttcaa caaagggtaa tatcgggaaa
11461 cctcctcgga ttccattgcc cagctatctg tcacttcatc aaaaggacag tagaaaagga
11521 aggtggcacc tacaaatgcc atcattgcga taaaggaaag gctatcgttc aagatgcctc
11581 tgccgacagt ggtcccaaag atggaccccc acccacgagg agcatcgtgg aaaaagaaga
11641 cgttccaacc acgtcttcaa agcaagtgga ttgatgtgat atctccactg acgtaaggga
11701 tgacgcacaa tcccactatc cttcgcaaga cccttcctct atataaggaa gttcatttca
11761 tttggagagg acacgctgaa atcaccagtc tctctctaca aatctatctc tctcgagctt
11821 tcgcagatcc ggggggcaat gagatatgaa aaagcctgaa ctcaccgcga cgtctgtcga
11881 gaagtttctg atcgaaaagt tcgacagcgt ctccgacctg atgcagctct cggagggcga
11941 agaatctcgt gctttcagct tcgatgtagg agggcgtgga tatgtcctgc gggtaaatag
12001 ctgcgccgat ggtttctaca aagatcgtta tgtttatcgg cactttgcat cggccgcgct
12061 cccgattccg gaagtgcttg acattgggga gtttagcgag agcctgacct attgcatctc
12121 ccgccgttca cagggtgtca cgttgcaaga cctgcctgaa accgaactgc ccgctgttct
12181 acaaccggtc gcggaggcta tggatgcgat cgctgcggcc gatcttagcc agacgagcgg
12241 gttcggccca ttcggaccgc aaggaatcgg tcaatacact acatggcgtg atttcatatg
12301 cgcgattgct gatccccatg tgtatcactg gcaaactgtg atggacgaca ccgtcagtgc
12361 gtccgtcgcg caggctctcg atgagctgat gctttgggcc gaggactgcc ccgaagtccg
12421 gcacctcgtg cacgcggatt tcggctccaa caatgtcctg acggacaatg gccgcataac
12481 agcggtcatt gactggagcg aggcgatgtt cggggattcc caatacgagg tcgccaacat
12541 cttcttctgg aggccgtggt tggcttgtat ggagcagcag acgcgctact tcgagcggag
12601 gcatccggag cttgcaggat cgccacgact ccgggcgtat atgctccgca ttggtcttga
12661 ccaactctat cagagcttgg ttgacggcaa tttcgatgat gcagcttggg cgcagggtcg
12721 atgcgacgca atcgtccgat ccggagccgg gactgtcggg cgtacacaaa tcgcccgcag
12781 aagcgcggcc gtctggaccg atggctgtgt agaagtactc gccgatagtg gaaaccgacg
12841 ccccagcact cgtccgaggg caaagaaata gagtagatgc cgaccgggat ctgtcgatcg
12901 acaagctcga gtttctccat aataatgtgt gagtagttcc cagataaggg aattagggtt
12961 cctatagggt ttcgctcatg tgttgagcat ataagaaacc cttagtatgt atttgtattt
13021 gtaaaatact tctatcaata aaatttctaa ttcctaaaac caaaatccag tactaaaatc
13081 cagatccccc gaattaattc ggcgttaatt cagtacatta aaaacgtccg caatgtgtta
13141 ttaagttgtc taagcgtcaa tttgtttaca ccacaatata tcctgccacc agccagccaa
13201 cagctccccg accggcagct cggcacaaaa tcaccactcg atacaggcag cccatcagtc
13261 cgggacggcg tcagcgggag agccgttgta aggcggcaga ctttgctcat gttaccgatg
13321 ctattcggaa gaacggcaac taagctgccg ggtttgaaac acggatgatc tcgcggaggg
13381 tagcatgttg attgtaacga tgacagagcg ttgctgcctg tgatcaccgc ggtttcaaaa
13441 tcggctccgt cgatactatg ttatacgcca actttgaaaa caactttgaa aaagctgttt
13501 tctggtattt aaggttttag aatgcaagga acagtgaatt ggagttcgtc ttgttataat
13561 tagcttcttg gggtatcttt aaatactgta gaaaagagga aggaaataat aaatggctaa
13621 aatgagaata tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac
13681 ggaaggaatg tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt
13741 aaaaatgacg gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat
13801 gatgctatgg ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga
13861 tggctggagc aatctgctca tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga
13921 agatgaacaa agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca
13981 ctccatcgac atatcggatt gtccctatac gaatagctta gacagccgct tagccgaatt
14041 ggattactta ctgaataacg atctggccga tgtggattgc gaaaactggg aagaagacac
14101 tccatttaaa gatccgcgcg agctgtatga ttttttaaag acggaaaagc ccgaagagga
14161 acttgtcttt tcccacggcg acctgggaga cagcaacatc tttgtgaaag atggcaaagt
14221 aagtggcttt attgatcttg ggagaagcgg cagggcggac aagtggtatg acattgcctt
14281 ctgcgtccgg tcgatcaggg aggatatcgg ggaagaacag tatgtcgagc tattttttga
14341 cttactgggg atcaagcctg attgggagaa aataaaatat tatattttac tggatgaatt
14401 gttttagtac ctagaatgca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
14461 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
14521 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
14581 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
14641 ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
14701 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
14761 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
14821 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
14881 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
14941 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
15001 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
15061 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
15121 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
15181 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
15241 gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg
15301 cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
15361 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
15421 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
15481 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
15541 gcgaggcagg gtgccttgat gtgggcgccg gcggtcgagt ggcgacggcg cggcttgtcc
15601 gcgccctggt agattgcctg gccgtaggcc agccattttt gagcggccag cggccgcgat
15661 aggccgacgc gaagcggcgg ggcgtaggga gcgcagcgac cgaagggtag gcgctttttg
15721 cagctcttcg gctgtgcgct ggccagacag ttatgcacag gccaggcggg ttttaagagt
15781 tttaataagt tttaaagagt tttaggcgga aaaatcgcct tttttctctt ttatatcagt
15841 cacttacatg tgtgaccggt tcccaatgta cggctttggg ttcccaatgt acgggttccg
15901 gttcccaatg tacggctttg ggttcccaat gtacgtgcta tccacaggaa acagaccttt
15961 tcgacctttt tcccctgcta gggcaatttg ccctagcatc tgctccgtac attaggaacc
16021 ggcggatgct tcgccctcga tcaggttgcg gtagcgcatg actaggatcg ggccagcctg
16081 ccccgcctcc tccttcaaat cgtactccgg caggtcattt gacccgatca gcttgcgcac
16141 ggtgaaacag aacttcttga actctccggc gctgccactg cgttcgtaga tcgtcttgaa
16201 caaccatctg gcttctgcct tgcctgcggc gcggcgtgcc aggcggtaga gaaaacggcc
16261 gatgccggga tcgatcaaaa agtaatcggg gtgaaccgtc agcacgtccg ggttcttgcc
16321 ttctgtgatc tcgcggtaca tccaatcagc tagctcgatc tcgatgtact ccggccgccc
16381 ggtttcgctc tttacgatct tgtagcggct aatcaaggct tcaccctcgg ataccgtcac
16441 caggcggccg ttcttggcct tcttcgtacg ctgcatggca acgtgcgtgg tgtttaaccg
16501 aatgcaggtt tctaccaggt cgtctttctg ctttccgcca tcggctcgcc ggcagaactt
16561 gagtacgtcc gcaacgtgtg gacggaacac gcggccgggc ttgtctccct tcccttcccg
16621 gtatcggttc atggattcgg ttagatggga aaccgccatc agtaccaggt cgtaatccca
16681 cacactggcc atgccggccg gccctgcgga aacctctacg tgcccgtctg gaagctcgta
16741 gcggatcacc tcgccagctc gtcggtcacg cttcgacaga cggaaaacgg ccacgtccat
16801 gatgctgcga ctatcgcggg tgcccacgtc atagagcatc ggaacgaaaa aatctggttg
16861 ctcgtcgccc ttgggcggct tcctaatcga cggcgcaccg gctgccggcg gttgccggga
16921 ttctttgcgg attcga
//
