| 質粒名稱: | pADH99 |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 90979 |
| 存儲實驗室: | Aaron Hernday |
|
載體骨架基本信息 |
|
| 載體名稱: | pUC19 |
| 載體出品公司: | Clontech |
| 空載體大小: | 2242bp |
| 完整質粒大小: | 11119bp |
| 載體類型: | CRISPR |
| 篩選標記: | Nourseothricin (NAT) |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷貝數: | 高拷貝 |
|
插入基因信息 |
|
| 插入基因名稱1: | US_HIS1 |
| 插入大小： | 433bp |
| 插入基因名稱2: | FRT |
| 插入大小： | 34bp |
| 插入基因名稱3: | ENO1 |
| 插入大小： | 783 bp |
| 插入基因名稱4: | C. albicans Cas9 |
| 插入大小： | 4113bp |
| 插入基因名稱5: | pMAL2 |
| 插入大小： | 548bp |
| 插入基因名稱6: | FLP |
| 插入大小： | 1272bp |
| 插入基因名稱7: | NAT 1 of 2 |
| 插入大小： | 846bp |
|
文獻引用方法 |
|
| 材料和方法部分: | pADH99 was a gift from Aaron Hernday (Addgene plasmid # 90979 ; //n2t.net/addgene:90979 ; RRID:Addgene_90979) |
| 參考文獻部分: | An Efficient, Rapid, and Recyclable System for CRISPR-Mediated Genome Editing in Candida albicans. Nguyen N, Quail MMF, Hernday AD. mSphere. 2017 Apr 26;2(2):e00149-17. doi: 10.1128/mSphereDirect.00149-17. eCollection 2017 Mar-Apr. 10.1128/mSphereDirect.00149-17 PubMed 28497115 |
買家導航
pADH99載體質粒基本信息
pADH99質粒圖譜載體圖譜和pADH99載體序列質粒序列多克隆位點信息
pADH99質粒載體簡介
pADH99質粒序列載體序列
LOCUS pADH99 11123 bp DNA circular SYN 14-APR-2025
DEFINITION NAT-marked CAS9 expression construct; part 1 of 2 of C.alb HIS1-FLP
CRISPR system. Use with pADH100-series gRNA expression constructs..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11123)
AUTHORS Nguyen N, Quail MMF, Hernday AD
TITLE An Efficient, Rapid, and Recyclable System for CRISPR-Mediated
Genome Editing in Candida albicans.
JOURNAL mSphere. 2017 Apr 26;2(2):e00149-17. doi:
10.1128/mSphereDirect.00149-17. eCollection 2017 Mar-Apr.
PUBMED 28497115
REFERENCE 2 (bases 1 to 11123)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1128/mSphereDirect.00149-17"; journalName: "mSphere"; date:
"2017-04-26- 26"; volume: "2"; issue: "2"; pages: "e00149-17"
COMMENT Sequence Label: pADH99
FEATURES Location/Qualifiers
source 1..11123
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 38..56
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
protein_bind complement(523..556)
/label=FRT (minimal)
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="supports FLP-mediated excision but not integration
(Turan and Bode, 2011)"
CDS 1344..5447
/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 1344..5447
/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 5460..5480
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5484..5504
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5508..5528
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5529..5594
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
terminator 5659..5906
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
primer_bind complement(5675..5693)
/label=CYC1
/note="CYC1 transcription termination signal, reverse
primer"
CDS 6465..7736
/codon_start=1
/product="site-specific recombinase"
/label=FLP
/note="FLP is a site-specific recombinase from
Saccharomyces cerevisiae. Recombination occurs at FRT
sequences."
/translation="MPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
YPAWNGIISQEVLDYLSSYINRRI"
promoter 8164..8507
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
promoter complement(join(9015..9021,9022..9039,9040..9045))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9060..9081
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9198..9215)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9369..9957)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9449..9468)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(10128..10919,10920..10988))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 10751..10770
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10989..11093)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt
61 tgacggcacg gccacgcgtt taaaccgcct caagccatgg gatatgctca aaatattgct
121 taactggggg tttaacaaat acatgtatgg taatccaaat gggtatgggg ttagtagtaa
181 tttggtagtg aaaaccccgg tagacggaga attaataaat accccaaaat ccaccacagg
241 actcgacagg tacctggagg atgaggagac agaagttagt agtaacaatt tggggacaga
301 agaggataca gtgacatcag aggaagaaga cagcgaaggt atagtcaaaa agcaatagga
361 cagcaagaca atttatttat ttaatagttt tttataaatg taaaatctat cacaacatgt
421 gtacttaatt ttctgtggta ggggaggcca tttcgttgaa acgtgtcgcg agcggttgtg
481 agaattgact cattttgatt attacgcaac cattgaaatt ttgaagttcc tatactttct
541 agagaatagg aacttcacag ctaacaatta cacaaaaaaa attctctttt cgatgtagca
601 actatcttca actaaaacat tttctccttc ggcccatgat tgtcctccgg gtcgacagca
661 agccgttaca attgagatgg aaagcgacct accttcactc gataaggtgc ttaattgtac
721 ttcatataaa tctggcccgg atctaaacaa atgagttcca ttaagccgtg ggttctcaat
781 tagggttttt gtttttgatt tagaaaaaag agatcaagat ttgtttacag gtgatgcctt
841 tttttagaac ttatgcgttg caaaagttga ctaacgattt ctataaggtg atccacacta
901 attatacaaa cgtacaaaca gacatacttt tcctgcgttc acctgatgtt ggccagattt
961 ctctcttcat tgcatagaac ataaccacac tagggcaaca gaaaaaaaaa aaaaaaaagt
1021 gcatcgggaa gttgtgttcc attcattata tgtctactac tgcatatgag tagcccaccc
1081 accaccacca tagtaagttt ttgtgtatgc gcgccgtcag gttatttcat ttctgaattt
1141 ttcaaccacc ttactccctt tattgttgat tgacaatttt gctcacagta agatctttta
1201 gactccaatt aatataaaat aagtctgatt ttccaattcc tgttttttct ttttttttct
1261 gtttctattt ctttcctttt ctcccttttt tttaattctt cattcaatca tcaattgata
1321 attcaggaat attacaaccc gggatggata aaaagtatag tattggttta gatattggta
1381 ctaactctgt gggttgggca gttatcactg acgaatataa agttccatca aagaaattta
1441 aggtgttagg taacactgac agacactcaa taaaaaagaa tcttatcggt gctcttttgt
1501 tcgactccgg tgaaactgcc gaggctacac gtttaaaaag aacagcaaga agaagatata
1561 cccgtagaaa aaatagaata tgttatttac aagaaatctt ttctaatgaa atggctaaag
1621 ttgatgattc ctttttccat agattggaag agtcattttt ggttgaagaa gacaaaaagc
1681 atgagagaca tccaatcttt gggaatatag ttgatgaagt ggcttaccat gaaaaatatc
1741 ctaccattta tcatttaaga aagaaattgg tagattcaac tgataaagct gaccttagat
1801 taatctattt agcacttgcc catatgatta aatttagagg tcattttttg attgaaggtg
1861 atttgaaccc agataattct gacgtggata aattatttat tcaattagtc caaacctaca
1921 accaattatt tgaggaaaat ccaattaatg ctagtggtgt cgatgccaaa gctatattat
1981 cagccagatt atcaaaatct agacgtttgg aaaatttgat tgcccaattg ccaggagaaa
2041 aaaagaatgg attatttgga aacttgatcg cattatcatt gggtttgaca ccaaatttta
2101 aatctaattt tgatttagct gaagatgcta aattacaatt atcaaaagac acctatgacg
2161 acgatttgga caatttactt gctcaaattg gtgatcaata tgcagatttg ttcttagctg
2221 ctaaaaactt atctgatgct attttgttgt ctgatatttt gagagtgaac acagaaataa
2281 ccaaagctcc attatcagca tctatgatca aacgttatga tgaacaccat caggatttga
2341 ctttattgaa agctttggtg agacaacaat tgccagagaa gtataaagaa atctttttcg
2401 atcaatctaa aaacgggtat gcaggttata ttgatggggg tgcctcccaa gaggaatttt
2461 acaaatttat aaaacctatt ttagaaaaga tggatgggac tgaggaactt ttggtcaaat
2521 tgaacagaga agatttgtta cgtaaacaga gaacttttga taatggtagt atacctcacc
2581 aaattcattt gggtgagttg catgcaattt taagaagaca agaagatttt tatccatttt
2641 taaaagataa tagagaaaaa atcgagaaaa ttttaacctt tagaattcca tactatgttg
2701 ggcctttggc tagaggtaat tcaagatttg cctggatgac acgtaaatca gaagaaacta
2761 ttaccccttg gaattttgaa gaggttgttg ataaaggagc atcagcacag agttttattg
2821 aaagaatgac caatttcgat aaaaacttac caaatgaaaa agttttacca aaacattcct
2881 tgttatacga atattttact gtttacaatg aacttacaaa ggttaaatat gttactgaag
2941 gtatgcgtaa gccagccttt ttatctggag aacagaaaaa ggcaatagtt gatttattgt
3001 ttaaaacaaa tagaaaagtt actgttaaac aattaaaaga agattacttt aagaaaattg
3061 aatgttttga ttcagttgaa atcagtggtg ttgaagacag atttaatgct agtttaggaa
3121 cttaccatga tttacttaaa attatcaaag ataaagattt cttggataac gaagaaaatg
3181 aagacatttt agaagacatt gttttaacct taactttatt cgaagataga gagatgattg
3241 aagaacgttt gaagacttat gcacatttgt ttgacgataa agtgatgaaa cagttgaaaa
3301 gaagacgtta tactggatgg ggtagattgt ctcgtaaatt gatcaatgga attagagata
3361 aacaaagtgg taaaactatc ttggactttt tgaaatctga cggatttgct aatagaaatt
3421 tcatgcaatt gatccacgac gatagtttga catttaaaga agacatccaa aaggcccaag
3481 tgagtgggca aggtgattca ttacatgaac atattgcaaa tttagccgga tctcctgcta
3541 ttaagaaagg gatattacaa actgttaaag ttgtggatga attagtgaaa gtaatgggaa
3601 gacataaacc tgaaaacatt gtcattgaga tggcaagaga aaatcaaact acacaaaaag
3661 gacagaaaaa tagtagagaa cgtatgaaaa gaatagaaga gggtattaaa gaattgggta
3721 gtcaaatatt gaaagaacac ccagtggaaa atacccagtt gcaaaatgaa aaattatatc
3781 tttactacct tcaaaatgga cgtgatatgt atgttgatca ggaattagat ataaatagac
3841 tttcagatta tgatgtagat catatagttc cacaatcttt cttgaaagat gattccatag
3901 acaataaagt attaactaga agtgataaaa atagaggtaa aagtgataat gtcccaagtg
3961 aggaagtcgt caaaaagatg aaaaattact ggcgtcaact tttgaatgct aaattaatta
4021 ctcaaagaaa atttgataat ttgactaaag cagaaagagg tgggctttct gaattagata
4081 aagccgggtt cattaaaaga caattggtcg aaactagaca aattactaaa catgttgccc
4141 aaattttaga ttcccgtatg aacactaagt atgacgaaaa tgataagtta atacgtgagg
4201 ttaaagtcat tactttaaaa tcaaaacttg tctctgattt cagaaaggat ttccaattct
4261 ataaagttag agaaattaat aattatcatc atgctcatga tgcatatttg aatgctgtag
4321 ttggaactgc tttaatcaag aaatacccta aattagaatc tgaatttgta tatggtgatt
4381 acaaagtcta tgatgttaga aagatgattg ctaaatcaga acaagaaatt ggtaaagcta
4441 cagctaaata cttcttttac tctaacatta tgaatttctt taaaacagaa attactttgg
4501 caaacggtga aattagaaaa agacctctta ttgaaacaaa tggtgagact ggagagatag
4561 tttgggacaa agggcgtgat ttcgctactg ttagaaaagt tttatcaatg ccacaagtta
4621 acattgtaaa gaaaacagag gttcaaactg gtggtttctc aaaagaaagt attttgccta
4681 aaagaaatag tgataaattg attgccagaa aaaaggattg ggatccaaag aaatatggtg
4741 gtttcgactc accaaccgta gcctattctg ttttggttgt ggcaaaggtt gaaaagggta
4801 aaagtaaaaa gcttaaatca gtaaaagaac ttttgggtat tacaataatg gaaagaagtt
4861 cctttgaaaa gaaccctatt gattttttgg aagctaaagg ttataaggaa gtaaagaagg
4921 acttaataat caaattgcct aaatattctt tatttgaatt agaaaatggg agaaaaagaa
4981 tgttggcttc tgctggagaa ttgcaaaagg gtaatgaatt agcattgcct tccaaatatg
5041 ttaacttctt gtatttagct tcacactatg aaaagttgaa agggtcacca gaagataacg
5101 agcaaaaaca attatttgtt gaacaacaca aacactactt agatgagatt atagaacaaa
5161 ttagtgaatt cagtaaaaga gtgatattag ctgatgcaaa tttagataaa gttttgtcag
5221 cctataacaa acatagagat aagccaatta gagaacaagc agaaaacatt attcacttat
5281 ttacccttac caatttagga gcacctgctg ctttcaagta ttttgataca acaattgatc
5341 gtaaaagata tacctcaaca aaagaagtct tagacgccac cttaattcat caatcaatca
5401 ctggattgta tgagacaaga attgatttgt ctcaattggg tggtgatgaa ggggctgatc
5461 ctaagaagaa aagaaaagtt gatccaaaga aaaagcgtaa ggtggatcct aagaaaaaga
5521 gaaaggttga ctacaaagac catgacggtg attataaaga tcatgacatc gactacaagg
5581 atgacgatga caagtgataa tgactgcaga gatccatcga cctgccgcca agctaattcc
5641 gggcgaattt ctgtcgagtc atgtaattag ttatgtcacg cttacattca cgccctcccc
5701 ccacatccgc tctaaccgaa aaggaaggag ttagacaacc tgaagtctag gtccctattt
5761 atttttttat agttatgtta gtattaagaa cgttatttat atttcaaatt tttctttttt
5821 ttctgtacag acgcgtgtac gcatgtaaca ttatactgaa aaccttgctt gagaaggttt
5881 tgggacgctc gaaggcttta atttgcggcc ccaataatga ttggtttgat atttttgtct
5941 agtaccatct gtaccattac acttaaatta tctttatatc tgtctaactc gactgtctgg
6001 atttcattga tgtagtcgta tgcatcgtta gttccaaaaa atattgtcat caatttgata
6061 ttggtttccg actctaaaat ttttggaaga atttgtctag cgtgctctga gttgtagcca
6121 ctgaaaccac ggttaataac atccaatttt cggatataca cattctgtaa tgctggatga
6181 aagccatact gggtacaact aaactgggtg atggagtcac cgaacaacac aaatttaccg
6241 tattccatga ttgctatggt tgagaatttt ttttttttct tgtcccacgc catttttcaa
6301 attatgcagt tgagaatgtt agtttttgtg tacaccccgt tcgctgaata tttcggaata
6361 attcaaagat tggggagtgg gggaggcgat agacgaagac acggtataaa aatgggcaaa
6421 attttcccca actttttgca gtggtttaac taataatcgt cgacatgcca caatttgata
6481 tattatgtaa aacaccacct aaggtgcttg ttcgtcagtt tgtggaaagg tttgaaagac
6541 cttcaggtga gaaaatagca ttatgtgctg ctgaactaac ctatttatgt tggatgatta
6601 cacataacgg aacagcaatc aagagagcca cattcatgag ctataatact atcataagca
6661 attcgttgag tttcgatatt gtcaataaat cactccagtt taaatacaag acgcaaaaag
6721 caacaatttt ggaagcctca ttaaagaaat tgattcctgc ttgggaattt acaattattc
6781 cttactatgg acaaaaacat caatctgata tcactgatat tgtaagtagt ttgcaattac
6841 agttcgaatc atcggaagaa gcagataagg gaaatagcca cagtaaaaaa atgcttaaag
6901 cacttctaag tgagggtgaa agcatctggg agatcactga gaaaatacta aattcgtttg
6961 agtatacttc gagatttaca aaaacaaaaa ctttatacca attcctcttc ctagctactt
7021 tcatcaattg tggaagattc agcgatatta agaacgttga tccgaaatca tttaaattag
7081 tccaaaataa gtatttggga gtaataatcc agtgtttagt gacagagaca aagacaagcg
7141 ttagtaggca catatacttc tttagcgcaa ggggtaggat cgatccactt gtatatttgg
7201 atgaattttt gaggaattct gaaccagtcc taaaacgagt aaataggacc ggcaattctt
7261 caagcaataa acaggaatac caattattaa aagataactt agtcagatcg tacaataaag
7321 ctttgaagaa aaatgcgcct tattcaatct ttgctataaa aaatggccca aaatctcaca
7381 ttggaagaca tttgatgacc tcatttcttt caatgaaggg cctaacggag ttgactaatg
7441 ttgtgggaaa ttggagcgat aagcgtgctt ctgccgtggc caggacaacg tatactcatc
7501 agataacagc aatacctgat cactacttcg cactagtttc tcggtactat gcatatgatc
7561 caatatcaaa ggaaatgata gcattgaagg atgagactaa tccaattgag gagtggcagc
7621 atatagaaca gctaaagggt agtgctgaag gaagcatacg ataccccgca tggaatggga
7681 taatatcaca ggaggtacta gactaccttt catcctacat aaatagacgc atataagagt
7741 gaaattctgg aaatctggaa atctggtttt gtattcttgt tattcttctt tttgttatta
7801 catatataac ttgttacttt tttaaaaaaa tctttgtata ttttataaat atataaaact
7861 aaatttaaga aaaagagaaa aatgttttat ttgagagatt gaaattttac ttgaatttag
7921 cttagctttt ataaagtatt attatgtaaa aaaacaaaac aaatatacat taaaaagtta
7981 agactataaa atagccaccc aaggcatttc tatatcttgt tgttgttgtt ttcatcttct
8041 gtatcagagg aacttatttt attattttcg tcacgggtat tttctcttgt ttgatgattc
8101 atcccattca ttccatcata aaatatcaag cttgcctcgt ccccgccggg tcacccggcc
8161 agcgacatgg aggcccagaa taccctcctt gacagtcttg acgtgcgcag ctcaggggca
8221 tgatgtgact gtcgcccgta catttagccc atacatcccc atgtataatc atttgcatcc
8281 atacattttg atggccgcac ggcgcgaagc aaaaattacg gctcctcgct gcagacctgc
8341 gagcagggaa acgctcccct cacagacgcg ttgaattgtc cccacgccgc gcccctgtag
8401 agaaatataa aaggttagga tttgccactg aggttcttct ttcatatact tccttttaaa
8461 atcttgctag gatacagttc tcacatcaca tccgaacata aacaaccgga tcctacaatt
8521 atgtctacta ctttggatga tactgcttat agatacagaa cttctgttcc aggtgatgct
8581 gaagctattg aagctttgga tggttctttc actaccgata ctgtttttag agttactgct
8641 actggtgatg gtttcacttt gagagaagtt ccagttgatc caccattgac taaggttttc
8701 ccagatgatg aatccgatga tgaatccgat gctggtgaag atggtgatcc agattctaga
8761 actttcgttg cttatggtga tgatggtgat ttggctggtt tcgttgttgt ttcttattct
8821 ggttggaaca gaagattgac tgttgaagat attgaagttg ctccagaaca tagaggtcat
8881 ggtgttggta gagctttgat gggtttggct actgaattcg ccagagaaag aggtgctggt
8941 catttgtggt tggaagttac caatgttaat cggtgtttaa accccagcgc ctggcgggcg
9001 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
9061 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
9121 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
9181 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
9241 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
9301 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
9361 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
9421 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
9481 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
9541 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
9601 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
9661 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
9721 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
9781 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
9841 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
9901 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
9961 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
10021 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
10081 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgcttg
10141 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
10201 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
10261 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
10321 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
10381 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
10441 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
10501 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
10561 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
10621 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
10681 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
10741 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
10801 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
10861 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
10921 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
10981 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
11041 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
11101 ccccgaaaag tgccacctga cgt
//
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