| 质粒名称: | pRS143 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 122377 |
| 存储实验室: | Rebecca Shapiro |
|
载体骨架基本信息 |
|
| 载体名称: | pJK-caCas9-NatMX-Neut5L |
| 载体出品公司: | -- |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 酵母表达，CRISPR |
| 筛选标记: | NAT |
| 载体抗性: | 氨苄青霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | -- |
|
插入基因信息 |
|
| 插入基因名称: | -- |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子: | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5’克隆位点: | -- |
| 3’克隆位点: | -- |
| 5’测序引物 | -- |
| 3’测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pRS143 was a gift from Rebecca Shapiro (Addgene plasmid # 122377 ; http://n2t.net/addgene:122377 ; RRID:Addgene_122377) |
| 参考文献部分: | A CRISPR Interference Platform for Efficient Genetic Repression in Candida albicans. Wensing L, Sharma J, Uthayakumar D, Proteau Y, Chavez A, Shapiro RS. mSphere. 2019 Feb 13;4(1). pii: 4/1/e00002-19. doi: 10.1128/mSphere.00002-19. 10.1128/mSphere.00002-19 PubMed 30760609 |
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 pRS143 12434 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION CRISPRi plasmid for use in Candida albicans. Contains dCas9 and
NEUT5L integration site and the sgRNA cloning site (SNR52 promoter,
PacI sites, sgRNA tail).
ACCESSION .
VERSION .
KEYWORDS pRS143
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12434)
AUTHORS Wensing L, Sharma J, Uthayakumar D, Proteau Y, Chavez A, Shapiro RS
TITLE A CRISPR Interference Platform for Efficient Genetic Repression in
Candida albicans.
JOURNAL mSphere. 2019 Feb 13;4(1). pii: 4/1/e00002-19. doi:
10.1128/mSphere.00002-19.
PUBMED 30760609
REFERENCE 2 (bases 1 to 12434)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..12434
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..31
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 39..55
/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)."
primer_bind 44..66
/label=M13/pUC Reverse
/note="In lacZ gene"
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"
promoter 100..118
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
CDS 1152..5255
/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 N863A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 5268..5288
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind 5293..5309
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 5294..5310
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
terminator 6246..6443
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
promoter complement(7044..7387)
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
primer_bind complement(8350..8369)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8351..8369)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8376..8393)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8376..8392)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8385..8407)
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind complement(8621..8640)
/label=F1ori-R
/note="F1 origin, reverse primer"
misc_RNA complement(8677..8751)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
primer_bind 10203..10224
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 10387..10491
/gene="bla"
/label=AmpR promoter
CDS 10492..11352
/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 complement(10710..10729)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 11523..12111
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 12012..12031
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 12265..12282
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 12399..12420
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
ORIGIN
1 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
61 cacaggaaac agctatgacc atgattacgc caagctcgaa attaaccctc actaaaggga
121 acaaaagctg gtaccagagc tattaagatc accagcctcg tttataataa acttagtcta
181 atatttcttt tggaccaaaa tagaaactgt tctcaggtgg caaaaaacag cttaaagtct
241 gtgttaaaaa aagatctcgg ctatgccaat caaaaaggag tgacattatt atgtaaaaga
301 agaaatgttg taattccgcg aaacaacaat tttaatgctt actgtgccgc caccaatatc
361 ctatggccaa ggggccttta aacatgccac ggaaatatac agtcaaagta aacctactat
421 gtctttagag ccttcaggat caattgtttt taaatgaatg actgcaaaac tagaaaagta
481 gcctcttaca taagcaagtc atttgtacac cgagagttta gtaaccagtg aagcaagtat
541 atctggcaac cctgaaactt atctgtgaat tataaagctc ataagtaaaa agtgtagaaa
601 agaagataag gcagaccaaa tacaaatttg caacagtaca acgaatattt cagcgtcaaa
661 actagagaat aataaagaaa acgatctttt caaaaagaaa aaacctttta gttttccttt
721 gttgttgttg tgggtgtgtg ctatttatat tatatagttt actcataata ccataaaata
781 ttcggtttga ttaggttatt ttaataagct aatttgtttc taatcgtgta atttatgctg
841 tgtatattaa gtagtgtgtg cactgcccaa aaatgtttgt tgtttatagt cggttaaaga
901 gaaaaaagaa aaaaagatcc atacacacac gttaattagt tgttcaacgt aatacactca
961 tattttgttc ttatttgctt tcggtcgctg ttctcaccaa gatttattgc caacgaaaca
1021 attttttttt atatattttc agatttttct ttttttcctt tcctttcctt ttctaatttt
1081 cactcctggt tttctttctt tcttagaaac attatctcga tattaatatt aaaaaaatat
1141 aatcattcaa aatggataaa aaatactcca taggtttggc cattggaacc aattcagttg
1201 gatgggctgt aatcactgat gaatataagg ttccttctaa aaagttcaaa gttttaggaa
1261 atactgatag acactccatt aaaaagaatt taattggtgc cttgttgttt gacagtggtg
1321 aaacagctga agcaactcgt ttgaaaagaa cagctagaag aagatatacc agaagaaaga
1381 acagaatttg ttacttgcaa gaaatctttt ctaatgaaat ggcaaaggtt gatgattctt
1441 tttttcatag attggaagaa agtttcttag ttgaagaaga taagaaacat gaaagacatc
1501 caatctttgg taatattgta gatgaagtgg cttaccacga aaaatatcct acaatttatc
1561 atttgagaaa aaaattagtt gactccactg ataaagctga cttaagactt atatatttgg
1621 cattggctca catgatcaag ttcagaggtc acttcttaat tgaaggtgat ttgaatccag
1681 ataattccga tgtcgacaag ttgtttattc aattggttca aacttacaac caacttttcg
1741 aggaaaatcc aatcaatgcc tcaggtgtgg atgctaaagc tattttatct gctagattgt
1801 caaaaagtag acgtttggaa aatcttattg cccaattgcc aggtgaaaaa aaaaatggat
1861 tgtttggaaa cttaattgca ttatctttgg gtttgacccc aaatttcaaa tcaaactttg
1921 atttggccga ggatgctaaa cttcaattgt ctaaggatac atacgatgac gatttagata
1981 atcttcttgc ccaaattggt gatcaatatg cagatttatt cttggcagct aagaacttga
2041 gtgatgcaat tttgttaagt gacattttga gagtcaacac agaaattaca aaagctcctt
2101 tatccgcctc tatgattaaa agatatgatg aacaccatca agatttaacc ttgttgaaag
2161 ctcttgtacg tcaacaatta ccagaaaagt ataaagaaat attctttgat caatctaaaa
2221 atggttacgc tggttatatt gatggtggtg ctagtcaaga agaattttat aagtttatca
2281 agccaattct tgaaaaaatg gatggtactg aggaattgtt ggttaagttg aatcgagagg
2341 atttacttcg taagcaaaga acatttgata acggttctat ccctcatcaa atccacttag
2401 gtgagttaca cgctatcttg agacgtcagg aagattttta cccttttttg aaagacaatc
2461 gtgaaaaaat tgaaaaaatt ttgactttca gaattcctta ttacgtcggg ccattggcta
2521 gaggtaattc tagatttgca tggatgactc gtaaatctga agagactatc accccatgga
2581 attttgagga agttgttgat aagggagcct ccgctcaatc attcatcgag agaatgacca
2641 atttcgacaa aaacttgcca aatgagaagg tgttacctaa acattctttg ttatatgaat
2701 atttcactgt ttataatgaa ttaacaaaag tcaaatacgt cactgaaggt atgagaaaac
2761 cagcattctt aagtggtgaa caaaagaaag ctatcgttga cttgttattc aaaactaatc
2821 gtaaagtcac tgtcaaacaa cttaaagagg actattttaa gaaaatagag tgtttcgatt
2881 ccgttgaaat cagtggggtt gaggatagat ttaatgctag tttaggaact tatcacgatt
2941 tgttgaaaat tatcaaagac aaagattttt tggataatga agaaaacgaa gatatcttgg
3001 aggacattgt attgactttg actttgtttg aggatagaga aatgatcgaa gaacgtttaa
3061 aaacttacgc tcacttattc gacgataaag tcatgaaaca attaaaaaga agaagatata
3121 ccggttgggg aagattatct agaaagttga ttaatggtat tagagataag caaagtggta
3181 aaaccatctt agatttcctt aaaagtgatg gttttgcaaa cagaaacttt atgcagttga
3241 ttcatgatga tagtttgacc tttaaggaag atattcaaaa agctcaggtt tctggacagg
3301 gtgattcatt gcatgaacat attgccaact tagcagggtc tccagctatc aaaaagggaa
3361 ttttacaaac tgttaaagtt gttgatgagt tggtgaaagt tatgggacgt cataagcctg
3421 aaaatatagt tatagaaatg gctagagaaa atcaaacaac tcagaaagga caaaagaatt
3481 ctcgtgaaag aatgaaaaga atagaagaag gaattaaaga gttgggttct caaatcttaa
3541 aagaacaccc agtcgaaaac acccagttac agaacgagaa gctttacttg tactatttgc
3601 aaaacgggcg tgacatgtat gtagatcaag agttggatat caatagactt tcagattacg
3661 atgtcgatca catagttcct cagtcatttt taaaagatga ttccattgac aacaaagttt
3721 taacaagatc tgataaggct agaggtaaat cagacaatgt gccatcagaa gaagtggtta
3781 agaaaatgaa gaactattgg agacaattgt taaatgcaaa gttgattact cagagaaaat
3841 ttgacaactt aactaaagct gaaagaggag gattatccga attagataaa gctggattca
3901 taaaacgtca gttggttgaa acaagacaaa ttactaaaca tgtagcccaa attttagatt
3961 cccgaatgaa tactaaatat gatgaaaatg acaaattaat cagagaggtc aaagtgatca
4021 cattaaagtc aaagttggtg tcagatttcc gaaaggattt tcaattctac aaggtgagag
4081 agatcaacaa ttatcatcat gctcacgacg catatttgaa tgcagtggta ggaactgcct
4141 taataaagaa gtatcctaag ttagaatccg aattcgtgta tggtgattat aaagtctacg
4201 atgtcagaaa gatgatagca aaatcagaac aagaaattgg gaaagccacc gcaaagtatt
4261 tcttttactc caacatcatg aatttcttta agaccgaaat caccttggct aacggggaaa
4321 tcagaaaacg tcctttgatt gaaacaaatg gagaaaccgg tgaaattgtg tgggataagg
4381 ggagagattt tgctactgtt agaaaagttt tatcaatgcc tcaagttaat attgtaaaga
4441 agactgaagt tcaaactggt ggtttctcta aggaatccat tcttccaaaa cgaaactcag
4501 acaagttgat cgcccgaaaa aaagattggg atcctaagaa atacggaggt tttgattctc
4561 ctactgtcgc atattctgtc ttggtcgtgg ctaaagttga aaagggtaaa tccaaaaaat
4621 tgaaatctgt gaaagaattg ttaggtatta ccatcatgga gcgttcatcc tttgagaaaa
4681 acccaattga ctttctcgaa gcgaagggct ataaagaagt gaagaaagat ttgattatta
4741 agttgccaaa gtattctctc tttgaacttg aaaatgggag aaagagaatg ttggcttcag
4801 ctggagaatt gcaaaaagga aacgagttag ctttgccttc aaagtacgtt aatttcttat
4861 acttagcatc tcattatgaa aagttgaagg ggtcccctga agacaatgag caaaagcaat
4921 tattcgtaga acaacataaa cactatttgg atgagataat tgaacaaatc tccgagtttt
4981 ctaaaagagt tattttggct gatgcaaatt tggataaagt tttatctgct tataataagc
5041 atcgtgacaa accaattcga gaacaagctg aaaatattat ccacttgttc acattgacta
5101 acttgggagc tccagccgcg tttaagtact ttgacaccac cattgaccgt aagagatata
5161 cttcaacaaa agaagtgtta gacgccacat tgattcatca gtccataact ggtttatatg
5221 aaactagaat cgatttgtca caattggggg gtgatagccg agcggatcct aagaaaaaac
5281 gaaaagtctg actcgaggtc gacggtatcg ataagcttga tatcgaattc ctgcagcccg
5341 ggggatccga gtgaaattct ggaaatctgg aaatctggtt ttgtattctt gttattcttc
5401 tttttgttat tacatatata acttgttact tttttaaaaa aatctttgtt tattttataa
5461 atatataaaa ctaaatttaa gaaaaagaga aaaatgtttt atttgagaga ttgaaatttt
5521 acttgaattt agcttagctt ttataaagta ttattatgta aaaaaacaaa acaaatatac
5581 attaaaaagt taagactata aaatagccac ccaaggcatt tctatatctt gttgttgttg
5641 ttttcatctt ctgtatcaga ggaacttatt ttattatttt cgtcacgggt attttctctt
5701 gtttgatgat tcatcccatt cattccatca taaaattcta gaaggaccac ctttgattgt
5761 aaatagtaat aattaccacc cttatttaat tatttattta acttatttat ttatttatta
5821 tacatatata caaatctaat aaagtgaaaa tctccccctt cacatttata attggccagt
5881 ttttttcaaa taagcattcc aaccagcatc tctatacctt ttaccttcaa tatctggatc
5941 tcttccttta ccaaacaatc ctctaccaac aatgataata tcagttccag tgctaacaac
6001 ttcatcaaca gttctatatt gttgtcctaa tccatcacct ttatcatcta atccaactcc
6061 aggtgtcata ataagccaat caaatccttc ttcttgtcca cccatatcac gttgggcaat
6121 aaatccaata acaaattcct tatcggattt agcaatttca acagtttttt gagaatattc
6181 tccatacgaa ttcgagctcg tttaaacgtc gacactggat ggcggcgtta gtatcgaatc
6241 gacagcagta tagcgaccag cattcacata cgattgacgc atgatattac tttctgcgca
6301 cttaacttcg catctgggca gatgatgtcg aggcgaaaaa aaatataaat cacgctaaca
6361 tttgattaaa atagaacaac tacaatataa aaaaactata caaatgacaa gttcttgaaa
6421 acaagaatct ttttattgtc agtactgact cgagttatta tggacatggc atagacatat
6481 acaaagcttg ttcaccatcg gaagcagtac catcgtataa agcagtatcc aaaccacaca
6541 aagtgaaacc cattcttcta taagcatgaa tagctggagc attaacattg gtaacttcca
6601 accacaaatg accagcacct ctttctctgg cgaattcagt agccaaaccc atcaaagctc
6661 taccaacacc atgacctcta tgttctggag caacttcaat atcttcaaca gtcaatcttc
6721 tgttccaacc agaataagaa acaacaacga aaccagccaa atcaccatca tcaccataag
6781 caacgaaagt tctagaatct ggatcaccat cttcaccagc atcggattca tcatcggatt
6841 catcatctgg gaaaacctta gtcaatggtg gatcaactgg aacttctctc aaagtgaaac
6901 catcaccagt agcagtaact ctaaaaacag tatcggtagt gaaagaacca tccaaagctt
6961 caatagcttc agcatcacct ggaacagaag ttctgtatct ataagcagta tcatccaaag
7021 tagtagacat aattgtagga tccggttgtt tatgttcgga tgtgatgtga gaactgtatc
7081 ctagcaagat tttaaaagga agtatatgaa agaagaacct cagtggcaaa tcctaacctt
7141 ttatatttct ctacaggggc gcggcgtggg gacaattcaa cgcgtctgtg aggggagcgt
7201 ttccctgctc gcaggtctgc agcgaggagc cgtaattttt gcttcgcgcc gtgcggccat
7261 caaaatgtat ggatgcaaat gattatacat ggggatgtat gggctaaatg tacgggcgac
7321 agtcacatca tgcccctgag ctgcgcacgt caagactgtc aaggagggta ttctgggcct
7381 ccatgtcgct ggccgggtga cccggcgggg acgaggcaag cttgatatgc taatgatccc
7441 actgatgata attcagctaa catcaataac cctcttggct cttggttggt ggtggtttct
7501 ttagctccct gttttaatcc ttcaactact ccattcccag tgacaccatg agcattagta
7561 atatctgccc aactactaat tttataaact ccaccaatat attgtttctt cacggtatta
7621 ccaatatcag caaattttct atcttcaaaa atcataaatt gatgtttacg tgaaagttct
7681 aataatggtt caatagtgga ttcataggaa aaatcattga ttatatcaat atgagtcttg
7741 attaagcata cataaggacc caatttatca attaattcaa ggaattcctt agtggtatca
7801 acatcaattg atgcacataa attggttttc ttcagttcca ttaatcgaaa taatcgttgt
7861 gctactggtg aggcatgagt ttctgctctc tcactatagg tcttagtgtt gactgtcata
7921 tcttttgttt gctatttcta tttctatttc tttcttcaac tgtaaaaggg gcgatatata
7981 aaccagtgtg tatggggttg ttgctcagga attgatcaat gtttaaaaga atggttgaaa
8041 tcatcattga agaacaaaaa aaaagatttt ttctttcatt atgttatctg tataatatgc
8101 aggaaataag attgcaaaaa aatagccgtt tttttggata ctatcaaaca agaggaatga
8161 ctaatacaga attagtgtta cgaatcaatg gcactacagc aactttcaac aaaataatct
8221 acgttcaaaa tataatatat actattaaat aaaatcaaat aataaaataa ttttaaagaa
8281 agtagtacca gctctttttt ttgtttccgt ttataccatc caaatcaatt cctattagtc
8341 tccaattcgc cctatagtga gtcgtattac aattcactgg ccgtcgtttt acaacgtcgt
8401 gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc ccctttcgcc
8461 agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg
8521 aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
8581 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
8641 tcctttctcg ccacgttcgc cggcaaaaaa aagcaccacc gactcggtgc cactttttca
8701 agttgataac ggactagcct tattttaact tgctatttct agctctaaaa ctgaagagcc
8761 attaatgctc ttcctcgaaa ttaaaaatag tttacgcaag tctcgaactt ccaccacaat
8821 ttaattgtaa gaacgctgcg aaaacatatt ctccaatcct attgggccga agtttcagct
8881 atgccaaagg catcaaattt ttttttataa aattttccac ttttccatac cagtgaaatt
8941 atcacgtgat ggagtgagta tctataatat ttgattgttt ctttgatgag gcaatactcg
9001 agtatagtta ttgaccaatg acttttaccg gctaaacaag tggtattcaa gcacaattca
9061 gttttctgga tctctaataa ttgcaattgc aattgcttca catactctgt ttataattca
9121 gtttctttct cttgatgtta ctgatatatt aagttaataa ataccaataa ctttttttca
9181 aaaatcttct attttccttt caattgatta tttaaacacg aatcgttaat aagctgtgat
9241 tgctttttct ctttcttttt cttttctttg ttcaagtttg aaaaaaaaaa aaaaaggttt
9301 ctcttgttcg atgtagaatt cagtgggtta tagttgtaat gatatatgga ataaaaagaa
9361 agaaagagaa gagaaagaga agaagagaag aattaagtaa acgtgtatta gaatttaaaa
9421 attaaaattc atgataatct ttaataataa ttaactaaga atcacaaaat tgaaaatttg
9481 aaagaaaaaa gatcactcca aggtcgtgtc aagtgactag aaattcttaa ttaaaatagt
9541 tataacaaaa aaagaaattt gtatataaca attaagtaaa agaaaaaaaa gcaattcgct
9601 ttgtataata atacccagaa aaatatattc atatattcat aaatcaaaca tgaaatgaag
9661 aatgctgaat cacttgatag gatttagttc cattatggta ctaagaagta atattacttc
9721 tatgaaaagg ttgaatccga gatcaaaatt catcttttca tatactctgg agaaaaaaaa
9781 aaaagtaatt gtagtaagaa tgacaagtat cagttgagaa aaaaagtaac aaagagaaag
9841 ctcggaggag gctccccaaa ggttttatca ccaatggtgg taccactaac ccaagaacag
9901 aaaaagcatt gatattttga aaaatacaat actcatcttt taacccaaac cttttcttgt
9961 cctttaaaac ttttatctca tgtctgtcca aagtcatatt gtcttactac tttctctcct
10021 tcaacgtcct tccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
10081 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
10141 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
10201 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
10261 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
10321 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
10381 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
10441 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
10501 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
10561 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
10621 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
10681 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
10741 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
10801 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
10861 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
10921 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
10981 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
11041 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
11101 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
11161 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
11221 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
11281 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
11341 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
11401 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
11461 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
11521 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
11581 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
11641 ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac
11701 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
11761 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
11821 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
11881 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
11941 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
12001 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
12061 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
12121 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
12181 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
12241 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgta agagcgccca
12301 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
12361 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
12421 taggcacccc aggc
//
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