| 质粒名称: | pMJ915 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 69090 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | pML-2CT |
| 载体出品公司: | MacroLab, UC Berkeley |
| 空载体大小: | 5950bp |
| 完整质粒大小: | 10094bp |
| 载体类型: | 细菌表达 |
| 载体抗性: | 氨苄霉素 |
| 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 |
| 物种来源: | Streptococcus pyogenes |
| 插入大小: | 5358 bp |
| 启动子: | T7 |
| 标签/融合蛋白: |
MBP (N terminal on backbone)
hexa His (N terminal on backbone)
two SV40 NLS (C terminal on insert)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | T7 promoter |
| 3’测序引物 |
7 terminator
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | pMJ915 was a gift from Jennifer Doudna (Addgene plasmid # 69090 ; http://n2t.net/addgene:69090 ; RRID:Addgene_69090) |
| 参考文献部分: | Enhanced homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery. Lin S, Staahl BT, Alla RK, Doudna JA. Elife. 2014 Dec 15;3:e04766. doi: 10.7554/eLife.04766. 10.7554/eLife.04766 PubMed 25497837 |
买家导航
pMJ915载体质粒基本信息
pMJ915质粒图谱载体图谱和pMJ915载体序列质粒序列多克隆位点信息
pMJ915质粒载体简介
pMJ915质粒序列载体序列
LOCUS pMJ915 10121 bp DNA circular SYN 14-APR-2025
DEFINITION Express Streptococcus pyogenes Cas9 carrying two C-terminal SV40
NLS.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10121)
AUTHORS Lin S, Staahl BT, Alla RK, Doudna JA
TITLE Enhanced homology-directed human genome engineering by controlled
timing of CRISPR/Cas9 delivery.
JOURNAL Elife. 2014 Dec 15;3:e04766. doi: 10.7554/eLife.04766.
PUBMED 25497837
REFERENCE 2 (bases 1 to 10121)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Elife.";
date: "2014-12-15"; volume: "3:e04766. doi"; pages: "
10.7554/eLife.04766"
COMMENT Sequence Label: pMJ915
FEATURES Location/Qualifiers
source 1..10121
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 1343..1362
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 1446..1465
/label=T7
/note="T7 promoter, forward primer"
promoter 1446..1464
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 1553..1575
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 1595..1612
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 1622..2722
/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="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 2696..2719
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 2777..2797
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
regulatory 2801..2810
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2807..6910
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 2807..6910
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 6917..6937
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6944..6964
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(7096..7114)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 7110..7157
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(join(7523..7528,7529..7545,7546..7551))
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(7578..7596)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7664..7768
/gene="bla"
/label=AmpR promoter
CDS join(7769..7837,7838..8629)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(7987..8006)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8800..9388
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9289..9308
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 9542..9559
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 9574..9713
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(9639..9658)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 9799..9821
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(9815..10006)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
ORIGIN
1 tgtgagggta aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt
61 caatgccagc gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct
121 gcgatgcaga tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac
181 gaaacacgga aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag
241 cagtcgcttc acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc
301 cgccagccta gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac
361 ccaacgctgc ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg
421 ttctgccaag ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt
481 cttggagtgg tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc
541 ggctccatgc accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa
601 tccatgccaa cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg
661 gtccaatgat cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat
721 ggtcgtcatc tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg
781 aagcgagaag aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga
841 cgtagcccag cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt
901 tggtggcggg accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa
961 gcgacaggcc gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga
1021 gcgctgccgg cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga
1081 cgatagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
1141 tcggtcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
1201 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
1261 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
1321 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
1381 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
1441 gaaattaata cgactcacta tagggagacc acaacggttt ccctctagtg ccggctccgg
1501 agagctcttt aattaagcgg ccgccctgca ggactcgagt tctagaaata attttgttta
1561 actttaagaa ggagatatac atatgaaatc ttctcaccat caccatcacc atggttcttc
1621 tatgaaaatc gaagaaggta aactggtaat ctggattaac ggcgataaag gctataacgg
1681 tctcgctgaa gtcggtaaga aattcgagaa agataccgga attaaagtca ccgttgagca
1741 tccggataaa ctggaagaga aattcccaca ggttgcggca actggcgatg gccctgacat
1801 tatcttctgg gcacacgacc gctttggtgg ctacgctcaa tctggcctgt tggctgaaat
1861 caccccggac aaagcgttcc aggacaagct gtatccgttt acctgggatg ccgtacgtta
1921 caacggcaag ctgattgctt acccgatcgc tgttgaagcg ttatcgctga tttataacaa
1981 agatctgctg ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg ataaagaact
2041 gaaagcgaaa ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact tcacctggcc
2101 gctgattgct gctgacgggg gttatgcgtt caagtatgaa aacggcaagt acgacattaa
2161 agacgtgggc gtggataacg ctggcgcgaa agcgggtctg accttcctgg ttgacctgat
2221 taaaaacaaa cacatgaatg cagacaccga ttactccatc gcagaagctg cctttaataa
2281 aggcgaaaca gcgatgacca tcaacggccc gtgggcatgg tccaacatcg acaccagcaa
2341 agtgaattat ggtgtaacgg tactgccgac cttcaagggt caaccatcca aaccgttcgt
2401 tggcgtgctg agcgcaggta ttaacgccgc cagtccgaac aaagagctgg caaaagagtt
2461 cctcgaaaac tatctgctga ctgatgaagg tctggaagcg gttaataaag acaaaccgct
2521 gggtgccgta gcgctgaagt cttacgagga agagttggcg aaagatccac gtattgccgc
2581 cactatggaa aacgcccaga aaggtgaaat catgccgaac atcccgcaga tgtccgcttt
2641 ctggtatgcc gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga
2701 agccctgaaa gacgcgcaga ctaattcgag ctcgaacaac aacaacaata acaataacaa
2761 caacctcggg atcgaggaaa acctgtactt ccaatccaat gccaccatgg ataagaaata
2821 ctcaataggc ttagatatcg gcacaaatag cgtcggatgg gcggtgatca ctgatgaata
2881 taaggttccg tctaaaaagt tcaaggttct gggaaataca gaccgccaca gtatcaaaaa
2941 aaatcttata ggggctcttt tatttgacag tggagagaca gcggaagcga ctcgtctcaa
3001 acggacagct cgtagaaggt atacacgtcg gaagaatcgt atttgttatc tacaggagat
3061 tttttcaaat gagatggcga aagtagatga tagtttcttt catcgacttg aagagtcttt
3121 tttggtggaa gaagacaaga agcatgaacg tcatcctatt tttggaaata tagtagatga
3181 agttgcttat catgagaaat atccaactat ctatcatctg cgaaaaaaat tggtagattc
3241 tactgataaa gcggatttgc gcttaatcta tttggcctta gcgcatatga ttaagtttcg
3301 tggtcatttt ttgattgagg gagatttaaa tcctgataat agtgatgtgg acaaactatt
3361 tatccagttg gtacaaacct acaatcaatt atttgaagaa aaccctatta acgcaagtgg
3421 agtagatgct aaagcgattc tttctgcacg attgagtaaa tcaagacgat tagaaaatct
3481 cattgctcag ctccccggtg agaagaaaaa tggcttattt gggaatctca ttgctttgtc
3541 attgggtttg acccctaatt ttaaatcaaa ttttgatttg gcagaagatg ctaaattaca
3601 gctttcaaaa gatacttacg atgatgattt agataattta ttggcgcaaa ttggagatca
3661 atatgctgat ttgtttttgg cagctaagaa tttatcagat gctattttac tttcagatat
3721 cctaagagta aatactgaaa taactaaggc tcccctatca gcttcaatga ttaaacgcta
3781 cgatgaacat catcaagact tgactctttt aaaagcttta gttcgacaac aacttccaga
3841 aaagtataaa gaaatctttt ttgatcaatc aaaaaacgga tatgcaggtt atattgatgg
3901 gggagctagc caagaagaat tttataaatt tatcaaacca attttagaaa aaatggatgg
3961 tactgaggaa ttattggtga aactaaatcg tgaagatttg ctgcgcaagc aacggacctt
4021 tgacaacggc tctattcccc atcaaattca cttgggtgag ctgcatgcta ttttgagaag
4081 acaagaagac ttttatccat ttttaaaaga caatcgtgag aagattgaaa aaatcttgac
4141 ttttcgaatt ccttattatg ttggtccatt ggcgcgtggc aatagtcgtt ttgcatggat
4201 gactcggaag tctgaagaaa caattacccc atggaatttt gaagaagttg tcgataaagg
4261 tgcttcagct caatcattta ttgaacgcat gacaaacttt gataaaaatc ttccaaatga
4321 aaaagtacta ccaaaacata gtttgcttta tgagtatttt acggtttata acgaattgac
4381 aaaggtcaaa tatgttactg aaggaatgcg aaaaccagca tttctttcag gtgaacagaa
4441 gaaagccatt gttgatttac tcttcaaaac aaatcgaaaa gtaaccgtta agcaattaaa
4501 agaagattat ttcaaaaaaa tagaatgttt tgatagtgtt gaaatttcag gagttgaaga
4561 tagatttaat gcttcattag gtacctacca tgatttgcta aaaattatta aagataaaga
4621 ttttttggat aatgaagaaa atgaagatat cttagaggat attgttttaa cattgacctt
4681 atttgaagat agggagatga ttgaggaaag acttaaaaca tatgctcacc tctttgatga
4741 taaggtgatg aaacagctta aacgtcgccg ttatactggt tggggacgtt tgtctcgaaa
4801 attgattaat ggtattaggg ataagcaatc tggcaaaaca atattagatt ttttgaaatc
4861 agatggtttt gccaatcgca attttatgca gctgatccat gatgatagtt tgacatttaa
4921 agaagacatt caaaaagcac aagtgtctgg acaaggcgat agtttacatg aacatattgc
4981 aaatttagct ggtagccctg ctattaaaaa aggtatttta cagactgtaa aagttgttga
5041 tgaattggtc aaagtaatgg ggcggcataa gccagaaaat atcgttattg aaatggcacg
5101 tgaaaatcag acaactcaaa agggccagaa aaattcgcga gagcgtatga aacgaatcga
5161 agaaggtatc aaagaattag gaagtcagat tcttaaagag catcctgttg aaaatactca
5221 attgcaaaat gaaaagctct atctctatta tctccaaaat ggaagagaca tgtatgtgga
5281 ccaagaatta gatattaatc gtttaagtga ttatgatgtc gatcacattg ttccacaaag
5341 tttccttaaa gacgattcaa tagacaataa ggtcttaacg cgttctgata aaaatcgtgg
5401 taaatcggat aacgttccaa gtgaagaagt agtcaaaaag atgaaaaact attggagaca
5461 acttctaaac gccaagttaa tcactcaacg taagtttgat aatttaacga aagctgaacg
5521 tggaggtttg agtgaacttg ataaagctgg ttttatcaaa cgccaattgg ttgaaactcg
5581 ccaaatcact aagcatgtgg cacaaatttt ggatagtcgc atgaatacta aatacgatga
5641 aaatgataaa cttattcgag aggttaaagt gattacctta aaatctaaat tagtttctga
5701 cttccgaaaa gatttccaat tctataaagt acgtgagatt aacaattacc atcatgccca
5761 tgatgcgtat ctaaatgccg tcgttggaac tgctttgatt aagaaatatc caaaacttga
5821 atcggagttt gtctatggtg attataaagt ttatgatgtt cgtaaaatga ttgctaagtc
5881 tgagcaagaa ataggcaaag caaccgcaaa atatttcttt tactctaata tcatgaactt
5941 cttcaaaaca gaaattacac ttgcaaatgg agagattcgc aaacgccctc taatcgaaac
6001 taatggggaa actggagaaa ttgtctggga taaagggcga gattttgcca cagtgcgcaa
6061 agtattgtcc atgccccaag tcaatattgt caagaaaaca gaagtacaga caggcggatt
6121 ctccaaggag tcaattttac caaaaagaaa ttcggacaag cttattgctc gtaaaaaaga
6181 ctgggatcca aaaaaatatg gtggttttga tagtccaacg gtagcttatt cagtcctagt
6241 ggttgctaag gtggaaaaag ggaaatcgaa gaagttaaaa tccgttaaag agttactagg
6301 gatcacaatt atggaaagaa gttcctttga aaaaaatccg attgactttt tagaagctaa
6361 aggatataag gaagttaaaa aagacttaat cattaaacta cctaaatata gtctttttga
6421 gttagaaaac ggtcgtaaac ggatgctggc tagtgccgga gaattacaaa aaggaaatga
6481 gctggctctg ccaagcaaat atgtgaattt tttatattta gctagtcatt atgaaaagtt
6541 gaagggtagt ccagaagata acgaacaaaa acaattgttt gtggagcagc ataagcatta
6601 tttagatgag attattgagc aaatcagtga attttctaag cgtgttattt tagcagatgc
6661 caatttagat aaagttctta gtgcatataa caaacataga gacaaaccaa tacgtgaaca
6721 agcagaaaat attattcatt tatttacgtt gacgaatctt ggagctcccg ctgcttttaa
6781 atattttgat acaacaattg atcgtaaacg atatacgtct acaaaagaag ttttagatgc
6841 cactcttatc catcaatcca tcactggtct ttatgaaaca cgcattgatt tgagtcagct
6901 aggaggtgac gggtcaccta agaaaaaacg aaaagttgag gatcctaaaa agaaacgaaa
6961 agttgattaa taacattgga agtggataac ggatccgcga tcgcggcgcg ccacctggtg
7021 gccggccggt accacgcgtg cgcgctgatc cggctgctaa caaagcccga aaggaagctg
7081 agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg
7141 tcttgagggg ttttttgctg aaaggaggaa ctatatccgg atatccacag gacgggtgtg
7201 gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg aagcgagcag gactgggcgg
7261 cggccaaagc ggtcggacag tgctccgaga acgggtgcgc atagaaattg catcaacgca
7321 tatagcgcta gcagcacgcc atagtgactg gcgatgctgt cggaatggac gatatcccgc
7381 aagaggcccg gcagtaccgg cataaccaag cctatgccta cagcatccag ggtgacggtg
7441 ccgaggatga cgatgagcgc attgttagat ttcatacacg gtgcctgact gcgttagcaa
7501 tttaactgtg ataaactacc gcattaaagc ttatcgatga taagctgtca aacatgagaa
7561 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
7621 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
7681 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
7741 gcttcaataa cattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
7801 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
7861 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
7921 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
7981 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
8041 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
8101 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
8161 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
8221 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
8281 accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact
8341 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
8401 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
8461 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
8521 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
8581 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
8641 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
8701 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
8761 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
8821 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
8881 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
8941 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
9001 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
9061 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
9121 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
9181 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
9241 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
9301 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
9361 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
9421 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
9481 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
9541 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
9601 tctgtgcggt atttcacacc gcaatggtgc actctcagta caatctgctc tgatgccgca
9661 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
9721 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
9781 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
9841 aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc gtgaagcgat tcacagatgt
9901 ctgcctgttc atccgcgtcc agctcgttga gtttctccag aagcgttaat gtctggcttc
9961 tgataaagcg ggccatgtta agggcggttt tttcctgttt ggtcactgat gcctccgtgt
10021 aagggggatt tctgttcatg ggggtaatga taccgatgaa acgagagagg atgctcacga
10081 tacgggttac tgatgatgaa catgcccggt tactggaacg t
//
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