| 质粒名称: | AIO-Puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 74630 |
| 存储实验室: | Steve Jackson |
|
载体骨架基本信息 |
|
| 载体名称: | All-in-One_Nickase-D10A |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 9626 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳动物表达,CRISPR |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
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 | Cbh |
| 标签/融合蛋白: | 嘌呤霉素抗性标记 |
| 克隆方法: | 限制性内切酶 |
| 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: | AIO-Puro was a gift from Steve Jackson (Addgene plasmid # 74630 ; http://n2t.net/addgene:74630 ; RRID:Addgene_74630) |
| 参考文献部分: | CRISPR-Cas9(D10A) nickase-based genotypic and phenotypic screening to enhance genome editing. Chiang TW, le Sage C, Larrieu D, Demir M, Jackson SP. Sci Rep. 2016 Apr 15;6:24356. doi: 10.1038/srep24356. 10.1038/srep24356 PubMed 27079678 |
买家导航
AIO-Puro载体 AIOPuro质粒基本信息
AIO-Puro质粒图谱 AIOPuro载体图谱和AIO-Puro载体序列 AIOPuro质粒序列多克隆位点信息
AIO-Puro质粒 AIOPuro载体简介
AIO-Puro质粒序列 AIOPuro载体序列
LOCUS AIO-Puro 9626 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION All-in-One plasmid encoding dual U6 promoter-driven sgRNAs and
Cas9-D10A nickase linked via 2A peptide with puromycin resistant
marker to enhance efficient and accurate genome editing.
ACCESSION .
VERSION .
KEYWORDS AIO-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9626)
AUTHORS Chiang TW, le Sage C, Larrieu D, Demir M, Jackson SP
TITLE CRISPR-Cas9(D10A) nickase-based genotypic and phenotypic screening
to enhance genome editing.
JOURNAL Sci Rep. 2016 Apr 15;6:24356. doi: 10.1038/srep24356.
PUBMED 27079678
REFERENCE 2 (bases 1 to 9626)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9626
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 7..247
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 283..358
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 448..465
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 483..723
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 758..833
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(842..860)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(843..860)
/label=SP6
/note="SP6 promoter, forward primer"
enhancer 897..1182
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 1184..1461
/label=chicken beta-actin promoter
intron 1462..1690
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1702..1711
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1711..1776
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1783..1803
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1828..5928
/codon_start=1
/product="nickase mutant of the Cas9 endonuclease from the
Streptococcus pyogenes Type II CRISPR/Cas system"
/label=Cas9(D10A)
/note="generates RNA-guided single strand nicks in DNA due
to the D10A mutation in the RuvC catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5929..5976
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5992..6048
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 6049..6645
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 6542..6562
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
primer_bind complement(6673..6690)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6679..6886
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6895..7035
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6895..7024
/label=AAV2 ITR
rep_origin 7110..7565
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7197..7216)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7407..7428
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7582..7601)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7701..7723
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7761..7779)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7847..7951
/gene="bla"
/label=AmpR promoter
CDS 7952..8812
/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(8170..8189)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8983..9571
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9472..9491
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 acatgtgagg gcctatttcc catgattcct tcatatttgc atatacgata caaggctgtt
61 agagagataa ttggaattaa tttgactgta aacacaaaga tattagtaca aaatacgtga
121 cgtagaaagt aataatttct tgggtagttt gcagttttaa aattatgttt taaaatggac
181 tatcatatgc ttaccgtaac ttgaaagtat ttcgatttct tggctttata tatcttgtgg
241 aaaggacgaa acaccggggt cttcggatcc atgcgaagac ctgttttaga gctagaaata
301 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
361 ttttgtttta gagctagaaa tagcaagtta aaataaggct agtccgtttt tagcgcgtgc
421 gccaattctg cagacaaatg gctctagatt aaccctcact aaagggaaag gtcgggcagg
481 aagagggcct atttcccatg attccttcat atttgcatat acgatacaag gctgttagag
541 agataattag aattaatttg actgtaaaca caaagatatt agtacaaaat acgtgacgta
601 gaaagtaata atttcttggg tagtttgcag ttttaaaatt atgttttaaa atggactatc
661 atatgcttac cgtaacttga aagtatttcg atttcttggc tttatatatc ttgtggaaag
721 gacgaaacac cgggagacca tcgatgagag ggtctcagtt ttagagctag aaatagcaag
781 ttaaaataag gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttt
841 ttctatagtg tcacctaaat tgcagctctg gcccgtgtct caaaatctct ggtacccgtt
901 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
961 tcaatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa
1021 actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc
1081 aatgacggta aatggcccgc ctggcattgt gcccagtaca tgaccttatg ggactttcct
1141 acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg
1201 ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt
1261 ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc gccaggcggg
1321 gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc agccaatcag
1381 agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg gccctataaa
1441 aagcgaagcg cgcggcgggc gggagtcgct gcgacgctgc cttcgccccg tgccccgctc
1501 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
1561 cgggcgggac ggcccttctc ctccgggctg taattagctg agcaagaggt aagggtttaa
1621 gggatggttg gttggtgggg tattaatgtt taattacctg gagcacctgc ctgaaatcac
1681 tttttttcag gttggaccgg tgccaccatg gactataagg accacgacgg agactacaag
1741 gatcatgata ttgattacaa agacgatgac gataagatgg ccccaaagaa gaagcggaag
1801 gtcggtatcc acggagtccc agcagccgac aagaagtaca gcatcggcct ggccatcggc
1861 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
1921 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
1981 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
2041 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
2101 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
2161 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
2221 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
2281 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
2341 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
2401 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
2461 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
2521 aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc
2581 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
2641 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
2701 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
2761 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
2821 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
2881 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
2941 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
3001 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
3061 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
3121 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
3181 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
3241 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
3301 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
3361 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
3421 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
3481 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
3541 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
3601 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
3661 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
3721 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
3781 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
3841 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
3901 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
3961 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
4021 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
4081 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
4141 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
4201 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
4261 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
4321 ctgtccgact acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc
4381 gacaacaagg tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc
4441 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
4501 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
4561 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
4621 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
4681 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
4741 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
4801 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
4861 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
4921 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
4981 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
5041 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
5101 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
5161 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
5221 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
5281 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
5341 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
5401 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
5461 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
5521 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
5581 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
5641 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
5701 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
5761 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
5821 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
5881 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacaa aaggccggcg
5941 gccacgaaaa aggccggcca ggcaaaaaag aaaaaggaat tcggctccgg agcaacaaac
6001 ttctctctgc tgaaacaagc cggagatgtc gaagagaatc ctggaccgac cgagtacaag
6061 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
6121 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
6181 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
6241 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
6301 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
6361 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
6421 gccaccgtcg gagtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
6481 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggaaac ctccgcgccc
6541 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
6601 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgagaatt ctaactagag
6661 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
6721 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
6781 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
6841 acagcaaggg ggaggattgg gaagagaata gcaggcatgc tggggagcgg ccgcaggaac
6901 ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact gaggccgggc
6961 gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc gagcgagcgc
7021 gcagctgcct gcaggggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt
7081 cacaccgcat acgtcaaagc aaccatagta cgcgccctgt agcggcgcat taagcgcggc
7141 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
7201 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
7261 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
7321 tgatttgggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
7381 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
7441 ccctatctcg ggctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
7501 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgtttac
7561 aattttatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagccccg
7621 acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg catccgctta
7681 cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac cgtcatcacc
7741 gaaacgcgcg agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
7801 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
7861 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
7921 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
7981 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
8041 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
8101 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
8161 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
8221 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
8281 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
8341 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
8401 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
8461 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
8521 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
8581 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
8641 tgataaatct ggagccggtg agcgtggaag ccgcggtatc attgcagcac tggggccaga
8701 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
8761 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
8821 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
8881 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
8941 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
9001 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
9061 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
9121 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
9181 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
9241 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
9301 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
9361 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
9421 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
9481 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
9541 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
9601 cctggccttt tgctggcctt ttgctc
//
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