| 质粒名称: | pMJ841 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 39318 |
| 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: | 卡那 25ug/mL |
| 空载体大小: | 6400 bp |
| 完整质粒大小: | 10500 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达,CRISPR |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9,Csn1 |
| 物种来源: | Streptococcus pyogenes |
| 插入大小: | 4104 bp |
| 基因突变: | 天冬氨酸10转化为丙氨酸（D10A）和组氨酸840转化为丙氨酸（H840A）；催化不活跃 |
| GenBank编号 | AAK33936.1 |
| 启动子 | T7 |
| 标签/融合蛋白: |
MBP（主干上的N终端）
His6（主干上的N终端）
|
| 克隆方法: | Ligation Independent克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | MBP-forward |
| 3' 测序引物 | T7 terminator |
|
文献引用方法 |
|
| 材料和方法部分: | pMJ841 was a gift from Jennifer Doudna (Addgene plasmid # 39318 ; http://n2t.net/addgene:39318 ; RRID:Addgene_39318) |
| 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 pMJ841 10577 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION synthetic circular DNA
ACCESSION .
VERSION .
KEYWORDS pMJ841
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"
primer_bind 30..49
/label=T7
/note="T7 promoter, forward primer"
promoter 30..48
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 49..73
/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 88..110
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 121..138
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 142..1239
/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 1213..1236
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 1270..1290
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQG"
CDS 1300..5406
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 5471..5488
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5541..5559)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5555..5602
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 5639..6094
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5726..5745)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5936..5957
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(6186..7001)
/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 6909..6928
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 6993..7012
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 7123..7711
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7612..7631
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7865..7882
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7897..8036
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7962..7981)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8122..8144
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(8138..8329)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
CDS complement(9138..10220)
/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 10182..10201
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(10221..10298)
/gene="lacI"
/label=lacI promoter
primer_bind 10504..10523
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 agaggatcga gatctcgatc ccgcgaaatt aatacgactc actatagggg aattgtgagc
61 ggataacaat tcccctctag aaataatttt gtttaacttt aagaaggaga tatacatatg
121 caccatcacc atcaccatgg aaaaatcgaa gaaggtaaac tggtaatctg gattaacggc
181 gataaaggct ataacggtct cgctgaagtc ggtaagaaat tcgagaaaga taccggaatt
241 aaagtcaccg ttgagcatcc ggataaactg gaagagaaat tcccacaggt tgcggcaact
301 ggcgatggcc ctgacattat cttctgggca cacgaccgct ttggtggcta cgctcaatct
361 ggcctgttgg ctgaaatcac cccggacaaa gcgttccagg acaagctgta tccgtttacc
421 tgggatgccg tacgttacaa cggcaagctg attgcttacc cgatcgctgt tgaagcgtta
481 tcgctgattt ataacaaaga tctgctgccg aacccgccaa aaacctggga agagatcccg
541 gcgctggata aagaactgaa agcgaaaggt aagagcgcgc tgatgttcaa cctgcaagaa
601 ccgtacttca cctggccgct gattgctgct gacgggggtt atgcgttcaa gtatgaaaac
661 ggcaagtacg acattaaaga cgtgggcgtg gataacgctg gcgcgaaagc gggtctgacc
721 ttcctggttg acctgattaa aaacaaacac atgaatgcag acaccgatta ctccatcgca
781 gaagctgcct ttaataaagg cgaaacagcg atgaccatca acggcccgtg ggcatggtcc
841 aacatcgaca ccagcaaagt gaattatggt gtaacggtac tgccgacctt caagggtcaa
901 ccatccaaac cgttcgttgg cgtgctgagc gcaggtatta acgccgccag tccgaacaaa
961 gagctggcaa aagagttcct cgaaaactat ctgctgactg atgaaggtct ggaagcggtt
1021 aataaagaca aaccgctggg tgccgtagcg ctgaagtctt acgaggaaga gttggcgaaa
1081 gatccacgta ttgccgccac tatggaaaac gcccagaaag gtgaaatcat gccgaacatc
1141 ccgcagatgt ccgctttctg gtatgccgtg cgtactgcgg tgatcaacgc cgccagcggt
1201 cgtcagactg tcgatgaagc cctgaaagac gcgcagacta attcgagctc gaacaacaac
1261 aacactagtg aaaacctgta tttccaggga gcagcctcga tggataagaa atactcaata
1321 ggcttagcta tcggcacaaa tagcgtcgga tgggcggtga tcactgatga atataaggtt
1381 ccgtctaaaa agttcaaggt tctgggaaat acagaccgcc acagtatcaa aaaaaatctt
1441 ataggggctc ttttatttga cagtggagag acagcggaag cgactcgtct caaacggaca
1501 gctcgtagaa ggtatacacg tcggaagaat cgtatttgtt atctacagga gattttttca
1561 aatgagatgg cgaaagtaga tgatagtttc tttcatcgac ttgaagagtc ttttttggtg
1621 gaagaagaca agaagcatga acgtcatcct atttttggaa atatagtaga tgaagttgct
1681 tatcatgaga aatatccaac tatctatcat ctgcgaaaaa aattggtaga ttctactgat
1741 aaagcggatt tgcgcttaat ctatttggcc ttagcgcata tgattaagtt tcgtggtcat
1801 tttttgattg agggagattt aaatcctgat aatagtgatg tggacaaact atttatccag
1861 ttggtacaaa cctacaatca attatttgaa gaaaacccta ttaacgcaag tggagtagat
1921 gctaaagcga ttctttctgc acgattgagt aaatcaagac gattagaaaa tctcattgct
1981 cagctccccg gtgagaagaa aaatggctta tttgggaatc tcattgcttt gtcattgggt
2041 ttgaccccta attttaaatc aaattttgat ttggcagaag atgctaaatt acagctttca
2101 aaagatactt acgatgatga tttagataat ttattggcgc aaattggaga tcaatatgct
2161 gatttgtttt tggcagctaa gaatttatca gatgctattt tactttcaga tatcctaaga
2221 gtaaatactg aaataactaa ggctccccta tcagcttcaa tgattaaacg ctacgatgaa
2281 catcatcaag acttgactct tttaaaagct ttagttcgac aacaacttcc agaaaagtat
2341 aaagaaatct tttttgatca atcaaaaaac ggatatgcag gttatattga tgggggagct
2401 agccaagaag aattttataa atttatcaaa ccaattttag aaaaaatgga tggtactgag
2461 gaattattgg tgaaactaaa tcgtgaagat ttgctgcgca agcaacggac ctttgacaac
2521 ggctctattc cccatcaaat tcacttgggt gagctgcatg ctattttgag aagacaagaa
2581 gacttttatc catttttaaa agacaatcgt gagaagattg aaaaaatctt gacttttcga
2641 attccttatt atgttggtcc attggcgcgt ggcaatagtc gttttgcatg gatgactcgg
2701 aagtctgaag aaacaattac cccatggaat tttgaagaag ttgtcgataa aggtgcttca
2761 gctcaatcat ttattgaacg catgacaaac tttgataaaa atcttccaaa tgaaaaagta
2821 ctaccaaaac atagtttgct ttatgagtat tttacggttt ataacgaatt gacaaaggtc
2881 aaatatgtta ctgaaggaat gcgaaaacca gcatttcttt caggtgaaca gaagaaagcc
2941 attgttgatt tactcttcaa aacaaatcga aaagtaaccg ttaagcaatt aaaagaagat
3001 tatttcaaaa aaatagaatg ttttgatagt gttgaaattt caggagttga agatagattt
3061 aatgcttcat taggtaccta ccatgatttg ctaaaaatta ttaaagataa agattttttg
3121 gataatgaag aaaatgaaga tatcttagag gatattgttt taacattgac cttatttgaa
3181 gatagggaga tgattgagga aagacttaaa acatatgctc acctctttga tgataaggtg
3241 atgaaacagc ttaaacgtcg ccgttatact ggttggggac gtttgtctcg aaaattgatt
3301 aatggtatta gggataagca atctggcaaa acaatattag attttttgaa atcagatggt
3361 tttgccaatc gcaattttat gcagctgatc catgatgata gtttgacatt taaagaagac
3421 attcaaaaag cacaagtgtc tggacaaggc gatagtttac atgaacatat tgcaaattta
3481 gctggtagcc ctgctattaa aaaaggtatt ttacagactg taaaagttgt tgatgaattg
3541 gtcaaagtaa tggggcggca taagccagaa aatatcgtta ttgaaatggc acgtgaaaat
3601 cagacaactc aaaagggcca gaaaaattcg cgagagcgta tgaaacgaat cgaagaaggt
3661 atcaaagaat taggaagtca gattcttaaa gagcatcctg ttgaaaatac tcaattgcaa
3721 aatgaaaagc tctatctcta ttatctccaa aatggaagag acatgtatgt ggaccaagaa
3781 ttagatatta atcgtttaag tgattatgat gtcgatgcca ttgttccaca aagtttcctt
3841 aaagacgatt caatagacaa taaggtctta acgcgttctg ataaaaatcg tggtaaatcg
3901 gataacgttc caagtgaaga agtagtcaaa aagatgaaaa actattggag acaacttcta
3961 aacgccaagt taatcactca acgtaagttt gataatttaa cgaaagctga acgtggaggt
4021 ttgagtgaac ttgataaagc tggttttatc aaacgccaat tggttgaaac tcgccaaatc
4081 actaagcatg tggcacaaat tttggatagt cgcatgaata ctaaatacga tgaaaatgat
4141 aaacttattc gagaggttaa agtgattacc ttaaaatcta aattagtttc tgacttccga
4201 aaagatttcc aattctataa agtacgtgag attaacaatt accatcatgc ccatgatgcg
4261 tatctaaatg ccgtcgttgg aactgctttg attaagaaat atccaaaact tgaatcggag
4321 tttgtctatg gtgattataa agtttatgat gttcgtaaaa tgattgctaa gtctgagcaa
4381 gaaataggca aagcaaccgc aaaatatttc ttttactcta atatcatgaa cttcttcaaa
4441 acagaaatta cacttgcaaa tggagagatt cgcaaacgcc ctctaatcga aactaatggg
4501 gaaactggag aaattgtctg ggataaaggg cgagattttg ccacagtgcg caaagtattg
4561 tccatgcccc aagtcaatat tgtcaagaaa acagaagtac agacaggcgg attctccaag
4621 gagtcaattt taccaaaaag aaattcggac aagcttattg ctcgtaaaaa agactgggat
4681 ccaaaaaaat atggtggttt tgatagtcca acggtagctt attcagtcct agtggttgct
4741 aaggtggaaa aagggaaatc gaagaagtta aaatccgtta aagagttact agggatcaca
4801 attatggaaa gaagttcctt tgaaaaaaat ccgattgact ttttagaagc taaaggatat
4861 aaggaagtta aaaaagactt aatcattaaa ctacctaaat atagtctttt tgagttagaa
4921 aacggtcgta aacggatgct ggctagtgcc ggagaattac aaaaaggaaa tgagctggct
4981 ctgccaagca aatatgtgaa ttttttatat ttagctagtc attatgaaaa gttgaagggt
5041 agtccagaag ataacgaaca aaaacaattg tttgtggagc agcataagca ttatttagat
5101 gagattattg agcaaatcag tgaattttct aagcgtgtta ttttagcaga tgccaattta
5161 gataaagttc ttagtgcata taacaaacat agagacaaac caatacgtga acaagcagaa
5221 aatattattc atttatttac gttgacgaat cttggagctc ccgctgcttt taaatatttt
5281 gatacaacaa ttgatcgtaa acgatatacg tctacaaaag aagttttaga tgccactctt
5341 atccatcaat ccatcactgg tctttatgaa acacgcattg atttgagtca gctaggaggt
5401 gactaacgag gccggtgctt tgcaggatcc gaattcgagc tccgtcgaca agcttgcggc
5461 cgcactcgag caccaccacc accaccactg agatccggct gctaacaaag cccgaaagga
5521 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
5581 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac
5641 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
5701 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
5761 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
5821 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
5881 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
5941 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
6001 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
6061 gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc
6121 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt
6181 aattcttaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
6241 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
6301 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
6361 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
6421 acgactgaat ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca
6481 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
6541 gattgcgcct gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
6601 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
6661 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
6721 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
6781 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
6841 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
6901 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
6961 cgcggcctag agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
7021 tttatgtaag cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt
7081 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
7141 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
7201 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
7261 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
7321 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
7381 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
7441 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
7501 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
7561 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
7621 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
7681 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
7741 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
7801 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
7861 gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac
7921 gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc
7981 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
8041 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt
8101 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
8161 cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc gattcacaga
8221 tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc
8281 ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg
8341 tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca
8401 cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac
8461 tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg
8521 ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga
8581 acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga
8641 agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc
8701 gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg
8761 tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg
8821 cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt
8881 gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt
8941 cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga
9001 agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg
9061 ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta
9121 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
9181 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt
9241 tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag
9301 agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg
9361 gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata
9421 tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga
9481 tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt
9541 tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg
9601 cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg
9661 cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc
9721 gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga
9781 aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc
9841 ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta
9901 caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg
9961 gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg
10021 gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg
10081 taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg
10141 gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg
10201 tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat
10261 gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt
10321 atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc
10381 cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc
10441 caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc gatcttcccc
10501 atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc
10561 cacgatgcgt ccggcgt
//
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