| 质粒名称: | pMJ826 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 39316 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pEC-K-MBP |
| 载体出品公司: | pET-based custom vector |
| 载体抗性: | Kan 25ug/mL |
| 空载体大小: | 6400 bp |
| 完整质粒大小: | 10500 bp |
| 载体类型: | 细菌表达，Crispr |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
| 宿生菌： | XL1 Blue |
| 生长说明： | 需要低卡那浓度，使用25ug/ml |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | 化脓性链球菌Streptococcus pyogenes |
| 备用名称： | Csn1 |
| 插入大小: | 4104 bp |
| 基因突变: | 组氨酸840到丙氨酸（H840A） |
| GenBank编号 | AAK33936.1 |
| 启动子 | T7 |
| 标签/融合蛋白: |
MBP (N terminal on backbone)
His6 (N terminal on backbone)
|
| 克隆方法: | 结扎独立克隆 |
| 5' 测序引物 | MBP转发 |
| 3' 测序引物 | T7端接器 |
|
文献引用方法 |
|
| 材料和方法部分: | pMJ826 was a gift from Jennifer Doudna (Addgene plasmid # 39316 ; http://n2t.net/addgene:39316 ; RRID:Addgene_39316) |
| 参考文献部分: | A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity. Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. Science. 2012 Jun 28. 10.1126/science.1225829 PubMed 22745249 |
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 pMJ826 10577 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMJ826
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10577)
AUTHORS Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E
TITLE A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive
Bacterial Immunity.
JOURNAL Science. 2012 Jun 28.
PUBMED 22745249
REFERENCE 2 (bases 1 to 10577)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10577
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 5..144
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(70..89)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 230..252
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(246..437)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(1246..2328)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 2290..2309
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(2329..2406)
/gene="lacI"
/label=lacI promoter
primer_bind 2612..2631
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 2715..2734
/label=T7
/note="T7 promoter, forward primer"
promoter 2715..2733
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 2734..2758
/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)."
RBS 2773..2795
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2806..2823
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2827..3924
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
YAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 3898..3921
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 3955..3975
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 3985..8091
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(H840A)
/note="generates RNA-guided single strand nicks in DNA due
to the H840A mutation in the HNH catalytic domain"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 8156..8173
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(8226..8244)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 8240..8287
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 8324..8779
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8411..8430)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8621..8642
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(8871..9686)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 9594..9613
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 9678..9697
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 9808..10396
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10297..10316
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10550..10567
/label=L4440
/note="L4440 vector, forward primer"
ORIGIN
1 agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcaatggtg
61 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac tccgctatcg
121 ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga cgcgccctga
181 cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc
241 atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg gtaaagctca
301 tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc cagctcgttg
361 agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt aagggcggtt
421 ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat gggggtaatg
481 ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg
541 ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa
601 atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc
661 cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt
721 tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac
781 gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca
841 gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc
901 cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt ggtggcggga
961 ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag cgacaggccg
1021 atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag cgctgccggc
1081 acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac gatagtcatg
1141 ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat cggtcgagat
1201 cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact gcccgctttc
1261 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
1321 ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg gcaacagctg
1381 attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc tggtttgccc
1441 cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg agctgtcttc
1501 ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg actcggtaat
1561 ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag tgggaacgat
1621 gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc agtcgccttc
1681 ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc cagccagacg
1741 cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct ggtgacccaa
1801 tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa taatactgtt
1861 gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc aggcagcttc
1921 cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac tgacgcgttg
1981 cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt ctaccatcga
2041 caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga caatttgcga
2101 cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact gtttgcccgc
2161 cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg cttccacttt
2221 ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa cggtctgata
2281 agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat tcaccaccct
2341 gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc gccattcgat
2401 ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag cagcccagta
2461 gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag gagatggcgc
2521 ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa gcgctcatga
2581 gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag gcgccagcaa
2641 ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg atcgagatct
2701 cgatcccgcg aaattaatac gactcactat aggggaattg tgagcggata acaattcccc
2761 tctagaaata attttgttta actttaagaa ggagatatac atatgcacca tcaccatcac
2821 catggaaaaa tcgaagaagg taaactggta atctggatta acggcgataa aggctataac
2881 ggtctcgctg aagtcggtaa gaaattcgag aaagataccg gaattaaagt caccgttgag
2941 catccggata aactggaaga gaaattccca caggttgcgg caactggcga tggccctgac
3001 attatcttct gggcacacga ccgctttggt ggctacgctc aatctggcct gttggctgaa
3061 atcaccccgg acaaagcgtt ccaggacaag ctgtatccgt ttacctggga tgccgtacgt
3121 tacaacggca agctgattgc ttacccgatc gctgttgaag cgttatcgct gatttataac
3181 aaagatctgc tgccgaaccc gccaaaaacc tgggaagaga tcccggcgct ggataaagaa
3241 ctgaaagcga aaggtaagag cgcgctgatg ttcaacctgc aagaaccgta cttcacctgg
3301 ccgctgattg ctgctgacgg gggttatgcg ttcaagtatg aaaacggcaa gtacgacatt
3361 aaagacgtgg gcgtggataa cgctggcgcg aaagcgggtc tgaccttcct ggttgacctg
3421 attaaaaaca aacacatgaa tgcagacacc gattactcca tcgcagaagc tgcctttaat
3481 aaaggcgaaa cagcgatgac catcaacggc ccgtgggcat ggtccaacat cgacaccagc
3541 aaagtgaatt atggtgtaac ggtactgccg accttcaagg gtcaaccatc caaaccgttc
3601 gttggcgtgc tgagcgcagg tattaacgcc gccagtccga acaaagagct ggcaaaagag
3661 ttcctcgaaa actatctgct gactgatgaa ggtctggaag cggttaataa agacaaaccg
3721 ctgggtgccg tagcgctgaa gtcttacgag gaagagttgg cgaaagatcc acgtattgcc
3781 gccactatgg aaaacgccca gaaaggtgaa atcatgccga acatcccgca gatgtccgct
3841 ttctggtatg ccgtgcgtac tgcggtgatc aacgccgcca gcggtcgtca gactgtcgat
3901 gaagccctga aagacgcgca gactaattcg agctcgaaca acaacaacac tagtgaaaac
3961 ctgtatttcc agggagcagc ctcgatggat aagaaatact caataggctt agatatcggc
4021 acaaatagcg tcggatgggc ggtgatcact gatgaatata aggttccgtc taaaaagttc
4081 aaggttctgg gaaatacaga ccgccacagt atcaaaaaaa atcttatagg ggctctttta
4141 tttgacagtg gagagacagc ggaagcgact cgtctcaaac ggacagctcg tagaaggtat
4201 acacgtcgga agaatcgtat ttgttatcta caggagattt tttcaaatga gatggcgaaa
4261 gtagatgata gtttctttca tcgacttgaa gagtcttttt tggtggaaga agacaagaag
4321 catgaacgtc atcctatttt tggaaatata gtagatgaag ttgcttatca tgagaaatat
4381 ccaactatct atcatctgcg aaaaaaattg gtagattcta ctgataaagc ggatttgcgc
4441 ttaatctatt tggccttagc gcatatgatt aagtttcgtg gtcatttttt gattgaggga
4501 gatttaaatc ctgataatag tgatgtggac aaactattta tccagttggt acaaacctac
4561 aatcaattat ttgaagaaaa ccctattaac gcaagtggag tagatgctaa agcgattctt
4621 tctgcacgat tgagtaaatc aagacgatta gaaaatctca ttgctcagct ccccggtgag
4681 aagaaaaatg gcttatttgg gaatctcatt gctttgtcat tgggtttgac ccctaatttt
4741 aaatcaaatt ttgatttggc agaagatgct aaattacagc tttcaaaaga tacttacgat
4801 gatgatttag ataatttatt ggcgcaaatt ggagatcaat atgctgattt gtttttggca
4861 gctaagaatt tatcagatgc tattttactt tcagatatcc taagagtaaa tactgaaata
4921 actaaggctc ccctatcagc ttcaatgatt aaacgctacg atgaacatca tcaagacttg
4981 actcttttaa aagctttagt tcgacaacaa cttccagaaa agtataaaga aatctttttt
5041 gatcaatcaa aaaacggata tgcaggttat attgatgggg gagctagcca agaagaattt
5101 tataaattta tcaaaccaat tttagaaaaa atggatggta ctgaggaatt attggtgaaa
5161 ctaaatcgtg aagatttgct gcgcaagcaa cggacctttg acaacggctc tattccccat
5221 caaattcact tgggtgagct gcatgctatt ttgagaagac aagaagactt ttatccattt
5281 ttaaaagaca atcgtgagaa gattgaaaaa atcttgactt ttcgaattcc ttattatgtt
5341 ggtccattgg cgcgtggcaa tagtcgtttt gcatggatga ctcggaagtc tgaagaaaca
5401 attaccccat ggaattttga agaagttgtc gataaaggtg cttcagctca atcatttatt
5461 gaacgcatga caaactttga taaaaatctt ccaaatgaaa aagtactacc aaaacatagt
5521 ttgctttatg agtattttac ggtttataac gaattgacaa aggtcaaata tgttactgaa
5581 ggaatgcgaa aaccagcatt tctttcaggt gaacagaaga aagccattgt tgatttactc
5641 ttcaaaacaa atcgaaaagt aaccgttaag caattaaaag aagattattt caaaaaaata
5701 gaatgttttg atagtgttga aatttcagga gttgaagata gatttaatgc ttcattaggt
5761 acctaccatg atttgctaaa aattattaaa gataaagatt ttttggataa tgaagaaaat
5821 gaagatatct tagaggatat tgttttaaca ttgaccttat ttgaagatag ggagatgatt
5881 gaggaaagac ttaaaacata tgctcacctc tttgatgata aggtgatgaa acagcttaaa
5941 cgtcgccgtt atactggttg gggacgtttg tctcgaaaat tgattaatgg tattagggat
6001 aagcaatctg gcaaaacaat attagatttt ttgaaatcag atggttttgc caatcgcaat
6061 tttatgcagc tgatccatga tgatagtttg acatttaaag aagacattca aaaagcacaa
6121 gtgtctggac aaggcgatag tttacatgaa catattgcaa atttagctgg tagccctgct
6181 attaaaaaag gtattttaca gactgtaaaa gttgttgatg aattggtcaa agtaatgggg
6241 cggcataagc cagaaaatat cgttattgaa atggcacgtg aaaatcagac aactcaaaag
6301 ggccagaaaa attcgcgaga gcgtatgaaa cgaatcgaag aaggtatcaa agaattagga
6361 agtcagattc ttaaagagca tcctgttgaa aatactcaat tgcaaaatga aaagctctat
6421 ctctattatc tccaaaatgg aagagacatg tatgtggacc aagaattaga tattaatcgt
6481 ttaagtgatt atgatgtcga tgccattgtt ccacaaagtt tccttaaaga cgattcaata
6541 gacaataagg tcttaacgcg ttctgataaa aatcgtggta aatcggataa cgttccaagt
6601 gaagaagtag tcaaaaagat gaaaaactat tggagacaac ttctaaacgc caagttaatc
6661 actcaacgta agtttgataa tttaacgaaa gctgaacgtg gaggtttgag tgaacttgat
6721 aaagctggtt ttatcaaacg ccaattggtt gaaactcgcc aaatcactaa gcatgtggca
6781 caaattttgg atagtcgcat gaatactaaa tacgatgaaa atgataaact tattcgagag
6841 gttaaagtga ttaccttaaa atctaaatta gtttctgact tccgaaaaga tttccaattc
6901 tataaagtac gtgagattaa caattaccat catgcccatg atgcgtatct aaatgccgtc
6961 gttggaactg ctttgattaa gaaatatcca aaacttgaat cggagtttgt ctatggtgat
7021 tataaagttt atgatgttcg taaaatgatt gctaagtctg agcaagaaat aggcaaagca
7081 accgcaaaat atttctttta ctctaatatc atgaacttct tcaaaacaga aattacactt
7141 gcaaatggag agattcgcaa acgccctcta atcgaaacta atggggaaac tggagaaatt
7201 gtctgggata aagggcgaga ttttgccaca gtgcgcaaag tattgtccat gccccaagtc
7261 aatattgtca agaaaacaga agtacagaca ggcggattct ccaaggagtc aattttacca
7321 aaaagaaatt cggacaagct tattgctcgt aaaaaagact gggatccaaa aaaatatggt
7381 ggttttgata gtccaacggt agcttattca gtcctagtgg ttgctaaggt ggaaaaaggg
7441 aaatcgaaga agttaaaatc cgttaaagag ttactaggga tcacaattat ggaaagaagt
7501 tcctttgaaa aaaatccgat tgacttttta gaagctaaag gatataagga agttaaaaaa
7561 gacttaatca ttaaactacc taaatatagt ctttttgagt tagaaaacgg tcgtaaacgg
7621 atgctggcta gtgccggaga attacaaaaa ggaaatgagc tggctctgcc aagcaaatat
7681 gtgaattttt tatatttagc tagtcattat gaaaagttga agggtagtcc agaagataac
7741 gaacaaaaac aattgtttgt ggagcagcat aagcattatt tagatgagat tattgagcaa
7801 atcagtgaat tttctaagcg tgttatttta gcagatgcca atttagataa agttcttagt
7861 gcatataaca aacatagaga caaaccaata cgtgaacaag cagaaaatat tattcattta
7921 tttacgttga cgaatcttgg agctcccgct gcttttaaat attttgatac aacaattgat
7981 cgtaaacgat atacgtctac aaaagaagtt ttagatgcca ctcttatcca tcaatccatc
8041 actggtcttt atgaaacacg cattgatttg agtcagctag gaggtgacta acgaggccgg
8101 tgctttgcag gatccgaatt cgagctccgt cgacaagctt gcggccgcac tcgagcacca
8161 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
8221 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
8281 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
8341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
8401 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
8461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
8521 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
8581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
8641 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
8701 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
8761 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
8821 gtttattttt ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac
8881 tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt
8941 tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca
9001 agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc
9061 ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt
9121 gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc
9181 tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg
9241 agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg
9301 cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat
9361 acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta
9421 cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc
9481 atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc
9541 gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga
9601 gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa
9661 gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac
9721 agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
9781 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
9841 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
9901 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
9961 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
10021 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
10081 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
10141 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
10201 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
10261 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
10321 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
10381 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
10441 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
10501 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
10561 gagcgaggaa gcggaag
//
