| 质粒名称: | pJK02 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 105133 |
| 存储实验室: | Joseph Sorg |
|
载体骨架基本信息 |
|
| 载体名称: | pMTL84151 |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 6297 bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Upstream & Downstream pyrE deletion region |
| 物种来源: | -- |
| 插入大小: | 2000 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pJK02 was a gift from Joseph Sorg (Addgene plasmid # 105133 ; http://n2t.net/addgene:105133 ; RRID:Addgene_105133) |
| 参考文献部分: | Using CRISPR-Cas9-mediated genome editing to generate C. difficile mutants defective in selenoproteins synthesis. McAllister KN, Bouillaut L, Kahn JN, Self WT, Sorg JA. Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5. 10.1038/s41598-017-15236-5 [pii] PubMed 29116155 |
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 pJK02 14173 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Clostridium difficile CRISPR-cas9 mutagenesis plasmid traJ oriT.
ACCESSION .
VERSION .
KEYWORDS pJK02
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14173)
AUTHORS McAllister KN, Bouillaut L, Kahn JN, Self WT, Sorg JA
TITLE Using CRISPR-Cas9-mediated genome editing to generate C. difficile
mutants defective in selenoproteins synthesis.
JOURNAL Sci Rep. 2017 Nov 7;7(1):14672. doi: 10.1038/s41598-017-15236-5.
PUBMED 29116155
REFERENCE 2 (bases 1 to 14173)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14173
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 63..79
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 63..79
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
CDS complement(2117..2737)
/codon_start=1
/gene="tetR from transposon Tn10"
/product="tetracycline repressor TetR"
/label=TetR
/note="TetR binds to the tetracycline operator tetO to
inhibit transcription. This inhibition can be relieved by
adding tetracycline or doxycycline."
/translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
primer_bind 2645..2664
/label=Tet-R
/note="Tetracycline resistance gene, reverse primer"
protein_bind 2847..2865
/label=tet operator
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O1 for the tetR and tetA genes"
CDS 2938..7044
/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"
primer_bind complement(7054..7071)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(7054..7070)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7063..7085)
/label=M13/pUC Forward
/note="In lacZ gene"
misc_RNA 12344..12419
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
rep_origin 12617..13205
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 13106..13125
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 13359..13376
/label=L4440
/note="L4440 vector, forward primer"
oriT 13526..13635
/note="incP origin of transfer"
CDS 13668..14039
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
ORIGIN
1 cctgcaggat aaaaaaattg tagataaatt ttataaaata gttttatcta caattttttt
61 atcaggaaac agctatgacc gcggccgcga cgtgattttt aatgggtaat ttaagtgtga
121 aatttggaag tgtagaattt aaaaatccag taataatggc gtctggaact tttggatttg
181 gaaaagaata taatgagatt tatgatatac aaaaacttgg tggtataagt agcaagggtc
241 ttactttaaa taaaaaacct ggtaacaatg gtatgagagt tcatgagaca ccatccggta
301 tgatgaatag tgttgggctt gaaaaccctg gagttcaagg atttatagac tatgaattac
361 ctttttttag taaactagac ttagtcagaa tagctaatgt tggaggtggg acattagaag
421 actacttatt aggggttcaa atgttaaatg acaagcctat agacattatt gaattaaata
481 tatcttgccc aaatgtcaaa gctggtggaa tggcttttgg tataaaaaat gaagtggcaa
541 gagaagttgt tcgtgaggta agaaacataa cgaaactgcc tttagtaata aaattatcac
601 ccaatgcaga agatattgta ggaatggcaa aagtttgtga agaagaaggt gctgatggag
661 tttcattagt taatactttt aaagcaatgg caattgatat aaaaaataga agaccagtat
721 ttgaaaatgt atatgctgga ttatctggac cagctataaa acctatagct ttgagaatgg
781 tacatgaagt gtgtaaaaat gtaaatatac ctgtaatggg aatgggcgga ataactaaag
841 ccacagatgc tatagagttt ataatggcag gagctacttg tatccaagtt ggaactgcaa
901 attttatgaa tcctagaata ggtattgaaa taatagatgg gattaatgaa tttatggata
961 gagaaggaat aaaaagttta gacgaaataa gaggaattat ataaggtaga ataaatagta
1021 ttttaaagat aaaaaatata gattaaataa acaataattt tataacatta acatggagga
1081 aaaataatta aaaacttaat tatttatagt gttacttaaa aaatgtaaat tttttagagt
1141 aaaaatttgt aatataacat ataacttaaa aataaaaagc agttagcctt ataaatgaag
1201 gttagctgtt ttttttgtta aaaatatatt tacaaaataa gagatagtag cagtacaata
1261 taagtataag gtaaaagtat tatatattag ggaggggtaa gtatgaaaaa tcaattaaat
1321 gaaaatcata aaaaggttgt attttttgga gtaggtgctg taggagctac ttttgcagaa
1381 caatttttta actctaaata tgattttaaa attctttgtg ataatgaaag aaagaaaaga
1441 tatttagaag aaggatttat aataaatgga aaaaggtatg attttaatta tgtaactaaa
1501 gatgagtata aacaagaggc tgattttata attataggtc taaaatataa taatttaaaa
1561 gaaaatataa aagaattaga tggattagtt ggaaaaaata cagttataat gtctctgcta
1621 aatggagttg atagcgaaga gataatagga gaaagatttg gaattggaaa aatggtatat
1681 tcatatgtta ccaatataga tgcaaagaaa atcaataata atattataca cactactaat
1741 gggataattg tatttggtaa caaagataat agtgaagata gaaaaactaa tataataact
1801 gaagtctttg atgatgtaaa tatagaatat attttatcaa aagatattca acgagatatg
1861 tggtggaagt acatggttaa tattggtgta aatcaaactt cagctatact tggtgcacct
1921 tatggagttt ttcagagttc tgagcattta agagaattag caaaatctgc aatgagggaa
1981 gttgttgcta tagcacaagc aaaagacata tctcttacag aagatgatgt tgaaaattca
2041 ttacatagaa tactagaaca ttcaaaagaa ggaagaacat caatgcttct cgaggcctgc
2101 agacatgcgc gatcgcagac ccactttcac atttaagttg tttttctaat ccgcatatga
2161 tcaattcaag gccgaataag aaggctggct ctgcaccttg gtgatcaaat aattcgatag
2221 cttgtcgtaa taatggcggc atactatcag tagtaggtgt ttccctttct tctttagcga
2281 cttgatgctc ttgatcttcc aatacgcaac ctaaagtaaa atgccccaca gcgctgagtg
2341 catataatgc attctctagt gaaaaacctt gttggcataa aaaggctaat tgattttcga
2401 gagtttcata ctgtttttct gtaggccgtg tacctaaatg tacttttgct ccatcgcgat
2461 gacttagtaa agcacatcta aaacttttag cgttattacg taaaaaatct tgccagcttt
2521 ccccttctaa agggcaaaag tgagtatggt gcctatctaa catctcaatg gctaaggcgt
2581 cgagcaaagc ccgcttattt tttacatgcc aatacaatgt aggctgctct acacctagct
2641 tctgggcgag tttacgggtt gttaaacctt cgattccgac ctcattaagc agctctaatg
2701 cgctgttaat cactttactt ttatctaatc tagacatcat taattcctcc tttttgttga
2761 cattatatca ttgatagagt tatttgtcaa actagttttt tatttcgatg ccctggactt
2821 catgaaaaac taaaaaaaat attgacactc tatcattgat agagtataat taaaataagc
2881 ttgatcgtag cgttaacaga tctgagctca ataatactag gaggtttttt taattaaatg
2941 gataaaaaat atagtatagg attagatata ggaacaaata gtgtaggatg ggcagtaata
3001 actgatgaat ataaagtacc aagtaaaaaa tttaaagtat taggaaatac agatagacat
3061 agtataaaaa aaaatttaat aggtgcatta ctttttgata gtggtgaaac agcagaagca
3121 acaagactta aaagaacagc aagaagaaga tatactagaa gaaaaaatag aatatgttat
3181 ttacaagaaa tatttagtaa tgaaatggca aaagtagatg atagtttttt tcatagatta
3241 gaagaaagtt ttttagtaga agaagataaa aaacacgaaa gacatccaat atttggaaat
3301 atagtagatg aagtagcata tcatgaaaaa tatccaacaa tatatcattt aagaaaaaaa
3361 cttgtagata gtacagataa agcagattta agacttatat atcttgcatt agcacatatg
3421 ataaaattta gaggacattt tcttatagaa ggtgatttaa atccagataa tagtgatgta
3481 gataaacttt ttatacaatt agtacaaaca tataatcaat tatttgaaga aaatccaata
3541 aatgcaagtg gtgtagatgc aaaagcaata ttaagtgcaa gattaagtaa aagtagaaga
3601 ttagaaaatc ttatagcaca attacctgga gaaaaaaaaa atggattatt tggtaatctt
3661 atagctttaa gtttaggatt aacaccaaat tttaaatcaa attttgatct tgcagaagat
3721 gcaaaacttc aattaagtaa agatacatat gatgatgatt tagataattt acttgcacaa
3781 ataggtgatc aatatgcaga tttatttctt gcagcaaaaa atctttctga tgcaatactt
3841 ttaagtgata tacttagagt aaatacagaa ataacaaaag caccattatc tgcaagtatg
3901 ataaaaagat atgatgaaca tcatcaagat ttaacattac ttaaagcatt agtaagacaa
3961 caattaccag aaaaatataa agaaattttt tttgatcaaa gtaaaaatgg atatgcagga
4021 tatatagatg gtggtgcaag tcaagaagaa ttttataaat ttattaaacc aatattagaa
4081 aaaatggatg gaacagaaga attattagta aaacttaata gagaagattt acttagaaaa
4141 caaagaacat ttgataatgg aagtatacca catcaaatac atttaggtga attacatgca
4201 atattaagaa gacaagaaga tttttatcca tttcttaaag ataatagaga aaaaattgaa
4261 aaaatactta catttagaat accatattat gtaggaccat tagcaagagg aaatagtaga
4321 tttgcttgga tgacaagaaa aagtgaagaa acaataacac catggaattt tgaagaagta
4381 gtagataaag gagcatctgc acaaagtttt atagaaagaa tgacaaattt tgataaaaat
4441 cttccaaatg aaaaagtatt accaaaacat agtttattat atgaatattt tacagtatat
4501 aatgaattaa caaaagtaaa atatgtaaca gaaggtatga gaaaaccagc atttttaagt
4561 ggtgaacaaa aaaaagctat agtagattta ttatttaaaa caaatagaaa agtaacagta
4621 aaacaattaa aagaagatta ttttaaaaaa attgaatgtt ttgatagtgt agaaatatct
4681 ggtgtagaag atagatttaa tgcaagttta ggaacatatc atgatttact taaaataata
4741 aaggataaag attttttaga taatgaagaa aatgaagata ttttagaaga tatagtatta
4801 acattaactt tatttgaaga tagagaaatg atagaagaaa gattaaaaac ttatgctcat
4861 ttatttgatg ataaagtaat gaaacaactt aaaagaagaa gatatacagg ttggggaaga
4921 ttaagtagaa aacttataaa tggaataaga gataaacaaa gtggaaaaac aatattagat
4981 tttcttaaaa gtgatggatt tgcaaataga aattttatgc aattaataca tgatgatagt
5041 ttaacattta aagaagatat tcaaaaagca caagtaagtg gacaaggtga tagtttacat
5101 gaacatatag caaatttagc aggatctcca gcaataaaaa aaggtatatt acaaacagta
5161 aaagtagttg atgaattagt aaaagtaatg ggtagacata aaccagaaaa tatagttata
5221 gaaatggcaa gagaaaatca aacaacacaa aaaggacaaa aaaatagtag agaaagaatg
5281 aaaagaatag aagaaggtat aaaagaatta ggaagtcaaa tacttaaaga acatccagta
5341 gaaaatacac aacttcaaaa tgaaaaactt tatttatatt atttacaaaa tggaagagat
5401 atgtatgtag atcaagaatt agatataaat agattaagtg attatgatgt agatcatata
5461 gtaccacaaa gttttcttaa agatgattct atagataata aagtattaac aagatctgat
5521 aaaaatagag gtaaaagtga taatgtacca agtgaagaag ttgtaaaaaa aatgaaaaat
5581 tattggagac aattacttaa tgcaaaactt ataacacaaa gaaaatttga taatcttaca
5641 aaagcagaaa gaggtggatt atctgaatta gataaagctg gatttataaa aagacaatta
5701 gtagaaacta gacaaataac aaaacatgta gcacaaatat tagatagtag aatgaataca
5761 aaatatgatg aaaatgataa gcttataaga gaagtaaaag ttataacatt aaaaagtaaa
5821 ttagtaagtg attttagaaa agattttcaa ttttataaag taagagaaat aaataattat
5881 catcatgcac atgatgcata tcttaatgca gtagtaggaa cagcattaat aaaaaaatat
5941 cctaaattag aatctgaatt tgtatatgga gattataaag tatatgatgt aagaaaaatg
6001 atagcaaaaa gtgaacaaga aataggaaaa gcaacagcaa agtacttttt ttattctaat
6061 ataatgaatt tttttaagac agaaataact ttagcaaatg gtgaaataag aaaaagacct
6121 ttaatagaaa ctaatggtga aacaggtgaa atagtatggg ataaaggtag agattttgca
6181 acagtaagaa aagtattaag tatgccacaa gtaaatatag taaaaaaaac agaagtacaa
6241 actggaggat ttagtaaaga aagtatactt ccaaaaagaa attcagataa acttattgca
6301 agaaaaaaag attgggatcc aaaaaaatat ggtggatttg atagtccaac agtagcatat
6361 agtgtattag tagtagcaaa agtagaaaaa ggtaaatcta aaaaacttaa aagtgtaaaa
6421 gaattacttg gaataacaat aatggaaaga agtagttttg aaaaaaatcc tattgatttt
6481 ttagaagcaa aaggatataa agaagttaaa aaagatttaa taataaaact tccaaaatat
6541 tctttatttg aattagaaaa tggaagaaaa agaatgttag caagtgctgg tgaattacaa
6601 aaaggtaatg aattagcatt accatctaaa tatgtaaatt ttctttattt agcaagtcat
6661 tatgaaaaac ttaaaggatc tcctgaagat aatgaacaaa aacaattatt tgtagaacaa
6721 cataaacatt atttagatga aataattgaa caaataagtg aattttctaa aagagtaata
6781 ttagcagacg caaatttaga taaagtatta tctgcttata ataaacatag agataaacca
6841 ataagagaac aagcagaaaa tataatacat ctttttacat taacaaattt aggtgcacca
6901 gcagcattta aatattttga tacaacaata gatagaaaaa gatatacaag tacaaaagaa
6961 gtattagacg caacattaat acatcaaagt ataacaggat tatatgaaac aagaatagat
7021 ttatctcaat taggtggtga ttaaaagctt ggcactggcc gtcgttttac aacgtcgtga
7081 ctgggaaaac cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag
7141 ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa
7201 tggcgaatgg cgctagcata aaaataagaa gcctgcattt gcaggcttct tatttttatg
7261 gcgcgcccgc ccttaagtct aaaaattagg ggagatgtaa ggatttggga aaaatagaag
7321 atgttataat cataaatatg gtattcgtag gcttaaagtc aaaaaggagg tgaaatataa
7381 atagattttt agctaaatta agtaagaaat aggaggagat ttattgaaca aaaaattaga
7441 aaaaccattt gtatataaga gagagtacga tttgactgga tatgatgttg aaattttaca
7501 aaaatatgag ttagaacaag caatatatgt ttatgttggg agtagttgtg catataacat
7561 gagagctaga agtagtaaat ggagatacca tataagaaca aataataagt ctatatgttg
7621 taacattaaa aattttatac ataacttgga attgttttat aaaatggaat taaagttgtc
7681 agataatatt attaatgata agctatacta tagcaatata gcagagtttg aagaatttga
7741 aacactagaa aaagctagag aggtagaaag tactattata agtcaatatc aatttttaga
7801 ttctataaat cacatgttaa aacaaaaaat aattttattg agtaataagg atagtgtgtt
7861 aaacataact aaaaatggaa atacaaatta tttgaaagta aaaaataaat acatagaaaa
7921 acataagaac aagccaataa tgagatacca tatcaactgt caattcaata cagatggaag
7981 tgtcaaaagt attacacagg agtttgaacc aatattggaa ttaaacaaaa aaaataccct
8041 aagccgacca agcagagtat ttttaaaata atattttaag ataacaacaa aatgagataa
8101 tactactaga caatgacaac tcaactacca attgagttta tggagctacc aactccaata
8161 tcggtctaac tgattaagta tctgtagtta tataataata ttgctatcaa ttttagcatc
8221 ttaacaatat tattatacat actaagctaa aattattcaa tagttgtaaa agttgattag
8281 tcaataagta tatatttaat gtagtgttat ctcttaaaaa aactagataa ggagataata
8341 aatatatgga acaattagat tcaaaatata agttgaaaaa atttctaatg gcagtattta
8401 gagatggtat aggacaagga aataatctta ttgataatga atatgttaga gtatttcaaa
8461 ataataaaag taatagtaaa caattagaac tcggagaaga atttaaagaa tatagtaaaa
8521 caactttttt taaaaatata gatgatatag tagaatttac cttcgcaaaa aatatttatt
8581 atgaaaatac attttttaac ctatgtacta ctgatggaaa agcaggaacc aatgaaaact
8641 taataaatag atatgcatta ggatttgatt ttgacaaaaa agaattagga caaggtttta
8701 attataaaga tataattaat ttatttacta agataggatt acattatcat atcctagttg
8761 atagtggaaa tggattccat gtttatgtgc taattaataa aactaataac attaagttag
8821 tatcagaagt tacaaataca ttaataaata aattgggtgc agataaacaa gcaaatttat
8881 ctactcaagt attaagagta ccttatacat ataatattaa aaatactact aaacaagtaa
8941 aaataataca ccaagacaaa aatatatata gatatgacat agaaaagtta gctaaaaaat
9001 attgcaaaga tgtaaaaaca gtaggtaata ctaatacaaa atatatatta gatagtaagc
9061 taccaaattg tatagtagat attttaaaaa atggtagtaa agatggacat aaaaacctag
9121 atttgcaaaa aatagttgtg actttaagat tgaggaataa aagtttaagt caagtaatat
9181 ccgttgctag agaatggaac tatatatcac aaaatagtct ttcaaatagt gagctagaat
9241 atcaagtcaa gtatatgtat gagaaactta aaacggttaa ttttggttgt actggttgtg
9301 agtttaatag tgattgttgg aataaaatag aatcagattt tatatatagt gatgaagata
9361 ctttgttcaa tatgccacat aagcactcaa aggatttgaa atataagaat aggaaagggg
9421 ttaaaataat gactggtaat caattgttta tctataatgt gttacttaac aataaagata
9481 gagaattaaa catagacgat ataatggagc tgataaccta taaacgtaag aagaaagtta
9541 aaaacattgt tatgagtgaa aagacattaa gagaaacatt aaaagaactt caacataatg
9601 attatattac aaaaacaaaa ggtgttacaa agctaggaat aaaagataca tacaatgtaa
9661 aagaagttag atgtaatata gataaacaat atactattag ttactttgtt accatggcag
9721 taatttgggg aataatttca actgaagaat taagattata tactcacatg agatataagc
9781 aagatttatt ggtcaaagat gataaaataa aaggaaatat attaagaatt aatcaagagg
9841 aattagcaaa agatttagga gtaacacagc aaagaatttc aaatatgata gaatctttat
9901 tagatactaa aattttagat gtatgggaaa ctaaaataaa tgatagagga tttatgtact
9961 atacatatag attaaacaag tagatttttg ataggattag aattgatttt ctagtcctat
10021 ttttatgcaa aaaaactaat taataaaaat ttcttttggt aaaataattg tacgagaatt
10081 gcaaaaaaaa aatggcatca aagtattgaa attaagccgt tttaaaaatt tcttttggta
10141 aaataattct acatatatat gtagtatata tatatatgtt ttttagagaa tgtataacta
10201 gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga atatagagaa
10261 tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata tagagctaga
10321 atatagagaa tgtataacta gaatatagag ctagaatata gagaatgtat aactagaata
10381 tagagctaga atatagagaa tgtataacta gaatatagag ctagaatcct aattagtagg
10441 tgctttttta aaacaagtta aaaatcaaaa atagtattag taagcattgg aaatgctaga
10501 ttctaaaata gaaaagtaaa aaattggtgc actatctaaa cttatctata tcgctttttc
10561 cgtcgtttgg ttctctagtt acgatacagg ggatatgctt atattgagtt atagtactaa
10621 tcagtgctta atatagttaa taaaattata gttaccatag tttagtaact atgatgtatg
10681 ttagttagaa acttgcattt cggccggcca gtgggcaagt tgaaaaattc acaaaaatgt
10741 ggtataatat ctttgttcat tagagcgata aacttgaatt tgagagggaa cttagatggt
10801 atttgaaaaa attgataaaa atagttggaa cagaaaagag tattttgacc actactttgc
10861 aagtgtacct tgtacctaca gcatgaccgt taaagtggat atcacacaaa taaaggaaaa
10921 gggaatgaaa ctatatcctg caatgcttta ttatattgca atgattgtaa accgccattc
10981 agagtttagg acggcaatca atcaagatgg tgaattgggg atatatgatg agatgatacc
11041 aagctataca atatttcaca atgatactga aacattttcc agcctttgga ctgagtgtaa
11101 gtctgacttt aaatcatttt tagcagatta tgaaagtgat acgcaacggt atggaaacaa
11161 tcatagaatg gaaggaaagc caaatgctcc ggaaaacatt tttaatgtat ctatgatacc
11221 gtggtcaacc ttcgatggct ttaatctgaa tttgcagaaa ggatatgatt atttgattcc
11281 tatttttact atggggaaat attataaaga agataacaaa attatacttc ctttggcaat
11341 tcaagttcat cacgcagtat gtgacggatt tcacatttgc cgttttgtaa acgaattgca
11401 ggaattgata aatagttaac ttcaggtttg tctgtaacta aaaacaagta tttaagcaaa
11461 aacatcgtag aaatacggtg ttttttgtta ccctaagttt cataaaaata agaagcctgc
11521 aaatgcaggc ttcttatttt tatggtttaa acggttttag ctgggatatc ggcaaaacaa
11581 gcaaatacta ctatagaagc agttaaaagg gctatttttt ataatctgga gcatagacaa
11641 gaagttagag aagcaattaa aaaagaaaat ataagaaaaa ttcttattat tgggacatca
11701 aaaaaaatgg ttaatcaaat tgtagagaga ttgtcaattg gaagggtctg taagtttata
11761 aatatacaag atataagtac taaatatgaa atagaaatag caaaacaagc tagaaaagaa
11821 ggaaatcatg taattcctgt tccagccgtt gagataaaaa gtatggcaag tggtctcagt
11881 ataaattctt tgaagagact ttttagaaaa ggaaataata ggaatatggt agttttagaa
11941 aaaacaatta taagaccaac ttttagttat ataggaaagt tttatattag tgctgatgta
12001 attaaacaga ttattgaata tgaagtatac aactttggaa gcattgatag aataaataaa
12061 atcaatatag agaattgcaa taattttatg aatatttttg taagtataaa tatcaatgat
12121 ttagaaacta taaaaaaaat tgaagacata caacagacaa taaaaaagtc tattgagaaa
12181 atgactttag ttaatgtaca aaaaatagat attaacattc ataaaattaa aaaaataaac
12241 ttcctccaat gttaaaaaat aaactgagaa aatgatatac taatttgcaa taaagtgtgc
12301 tataattaaa ctgtaaaggt accgaaaagt gatgcattgt tgggttttag agctagaaat
12361 agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct
12421 ttttttctat ggagaaatct agatcagcat gatgtctgac tagacgcgta agctctgcaa
12481 ctatttttag atggttgcag agcttatttt tattttttat taaactcctt tttgataatc
12541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
12601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
12661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
12721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
12781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
12841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
12901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
12961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
13021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
13081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
13141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
13201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
13261 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
13321 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
13381 cggaagagcg cccaatacgc agggccccct gcttcggggt cattatagcg attttttcgg
13441 tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt gtagactttc
13501 cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca cccgcgagcg
13561 ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg aatcctgctc
13621 tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg gctgatgaaa
13681 ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca gacgaacgaa
13741 gagcgattga ggaaaaggcg gcggcggccg gcatgagcct gtcggcctac ctgctggccg
13801 tcggccaggg ctacaaaatc acgggcgtcg tggactatga gcacgtccgc gagctggccc
13861 gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa actctggctc accgacgacc
13921 cgcgcacggc gcggttcggt gatgccacga tcctcgccct gctggcgaag atcgaagaga
13981 agcaggacga gcttggcaag gtcatgatgg gcgtggtccg cccgagggca gagccatgac
14041 ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg ccaagcacgt ccccatgcgc
14101 tccatcaaga agagcgactt cgcggagctg gtgaagtaca tcaccgacga gcaaggcaag
14161 accgatcggg ccc
//
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