| 质粒名称: | pU6-(BbsI)_CBh-Cas9-T2A-mCherry |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 64324 |
| 存储实验室: | Ralf Kuehn |
|
载体骨架基本信息 |
|
| 载体名称: | pX330 |
| 载体出品公司: | Zhang lab (Addgene plasmid # 42230) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 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: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Cas9 |
| 物种来源: | -- |
| 插入大小: | 4500 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: | pU6-(BbsI)_CBh-Cas9-T2A-mCherry was a gift from Ralf Kuehn (Addgene plasmid # 64324 ; http://n2t.net/addgene:64324 ; RRID:Addgene_64324) |
| 参考文献部分: | Increasing the efficiency of homology-directed repair for CRISPR-Cas9-induced precise gene editing in mammalian cells. Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, Kuhn R. Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198. 10.1038/nbt.3198 PubMed 25803306 |
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
_CBh-Cas9-T2A-mCherry质粒图谱.png)
载体简介
载体序列
LOCUS pU6-(BbsI)_CBh-Cas9-T2A-mCherry 9283 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expression vector for sgRNAs cloned into the BbsI sites and for
expression of Cas9 linked to mCherry via a T2A peptide.
ACCESSION .
VERSION .
KEYWORDS pU6-(BbsI)_CBh-Cas9-T2A-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9283)
AUTHORS Chu VT, Weber T, Wefers B, Wurst W, Sander S, Rajewsky K, Kuhn R
TITLE Increasing the efficiency of homology-directed repair for
CRISPR-Cas9-induced precise gene editing in mammalian cells.
JOURNAL Nat Biotechnol. 2015 Mar 24. doi: 10.1038/nbt.3198.
PUBMED 25803306
REFERENCE 2 (bases 1 to 9283)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9283
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 7..247
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 7..27
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 178..197
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 274..349
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 446..731
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 733..1010
/label=chicken beta-actin promoter
intron 1011..1239
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1251..1260
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1260..1325
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1332..1352
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1377..5477
/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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5478..5525
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 5529..5582
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5589..6299
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(5734..5752)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind complement(6324..6341)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6330..6537
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 6546..6686
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 6546..6675
/label=AAV2 ITR
rep_origin 6761..7216
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6848..6867)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7058..7079
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7233..7252)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7352..7374
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(7412..7430)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7498..7602
/gene="bla"
/label=AmpR promoter
CDS 7603..8463
/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(7821..7840)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 8634..9222
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9123..9142
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 ccccacgagg 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 acaccgggtc ttcgagaaga cctgttttag agctagaaat agcaagttaa
301 aataaggcta gtccgttatc aacttgaaaa agtggcaccg agtcggtgct tttttgtttt
361 agagctagaa atagcaagtt aaaataaggc tagtccgttt ttagcgcgtg cgccaattct
421 gcagacaaat ggctctagag gtacccgtta cataacttac ggtaaatggc ccgcctggct
481 gaccgcccaa cgacccccgc ccattgacgt caatagtaac gccaataggg actttccatt
541 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
601 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattgtg
661 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
721 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
781 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
841 gggggggggg ggggcgcgcg ccaggcgggg cggggcgggg cgaggggcgg ggcggggcga
901 ggcggagagg tgcggcggca gccaatcaga gcggcgcgct ccgaaagttt ccttttatgg
961 cgaggcggcg gcggcggcgg ccctataaaa agcgaagcgc gcggcgggcg ggagtcgctg
1021 cgacgctgcc ttcgccccgt gccccgctcc gccgccgcct cgcgccgccc gccccggctc
1081 tgactgaccg cgttactccc acaggtgagc gggcgggacg gcccttctcc tccgggctgt
1141 aattagctga gcaagaggta agggtttaag ggatggttgg ttggtggggt attaatgttt
1201 aattacctgg agcacctgcc tgaaatcact ttttttcagg ttggaccggt gccaccatgg
1261 actataagga ccacgacgga gactacaagg atcatgatat tgattacaaa gacgatgacg
1321 ataagatggc cccaaagaag aagcggaagg tcggtatcca cggagtccca gcagccgaca
1381 agaagtacag catcggcctg gacatcggca ccaactctgt gggctgggcc gtgatcaccg
1441 acgagtacaa ggtgcccagc aagaaattca aggtgctggg caacaccgac cggcacagca
1501 tcaagaagaa cctgatcgga gccctgctgt tcgacagcgg cgaaacagcc gaggccaccc
1561 ggctgaagag aaccgccaga agaagataca ccagacggaa gaaccggatc tgctatctgc
1621 aagagatctt cagcaacgag atggccaagg tggacgacag cttcttccac agactggaag
1681 agtccttcct ggtggaagag gataagaagc acgagcggca ccccatcttc ggcaacatcg
1741 tggacgaggt ggcctaccac gagaagtacc ccaccatcta ccacctgaga aagaaactgg
1801 tggacagcac cgacaaggcc gacctgcggc tgatctatct ggccctggcc cacatgatca
1861 agttccgggg ccacttcctg atcgagggcg acctgaaccc cgacaacagc gacgtggaca
1921 agctgttcat ccagctggtg cagacctaca accagctgtt cgaggaaaac cccatcaacg
1981 ccagcggcgt ggacgccaag gccatcctgt ctgccagact gagcaagagc agacggctgg
2041 aaaatctgat cgcccagctg cccggcgaga agaagaatgg cctgttcgga aacctgattg
2101 ccctgagcct gggcctgacc cccaacttca agagcaactt cgacctggcc gaggatgcca
2161 aactgcagct gagcaaggac acctacgacg acgacctgga caacctgctg gcccagatcg
2221 gcgaccagta cgccgacctg tttctggccg ccaagaacct gtccgacgcc atcctgctga
2281 gcgacatcct gagagtgaac accgagatca ccaaggcccc cctgagcgcc tctatgatca
2341 agagatacga cgagcaccac caggacctga ccctgctgaa agctctcgtg cggcagcagc
2401 tgcctgagaa gtacaaagag attttcttcg accagagcaa gaacggctac gccggctaca
2461 ttgacggcgg agccagccag gaagagttct acaagttcat caagcccatc ctggaaaaga
2521 tggacggcac cgaggaactg ctcgtgaagc tgaacagaga ggacctgctg cggaagcagc
2581 ggaccttcga caacggcagc atcccccacc agatccacct gggagagctg cacgccattc
2641 tgcggcggca ggaagatttt tacccattcc tgaaggacaa ccgggaaaag atcgagaaga
2701 tcctgacctt ccgcatcccc tactacgtgg gccctctggc caggggaaac agcagattcg
2761 cctggatgac cagaaagagc gaggaaacca tcaccccctg gaacttcgag gaagtggtgg
2821 acaagggcgc ttccgcccag agcttcatcg agcggatgac caacttcgat aagaacctgc
2881 ccaacgagaa ggtgctgccc aagcacagcc tgctgtacga gtacttcacc gtgtataacg
2941 agctgaccaa agtgaaatac gtgaccgagg gaatgagaaa gcccgccttc ctgagcggcg
3001 agcagaaaaa ggccatcgtg gacctgctgt tcaagaccaa ccggaaagtg accgtgaagc
3061 agctgaaaga ggactacttc aagaaaatcg agtgcttcga ctccgtggaa atctccggcg
3121 tggaagatcg gttcaacgcc tccctgggca cataccacga tctgctgaaa attatcaagg
3181 acaaggactt cctggacaat gaggaaaacg aggacattct ggaagatatc gtgctgaccc
3241 tgacactgtt tgaggacaga gagatgatcg aggaacggct gaaaacctat gcccacctgt
3301 tcgacgacaa agtgatgaag cagctgaagc ggcggagata caccggctgg ggcaggctga
3361 gccggaagct gatcaacggc atccgggaca agcagtccgg caagacaatc ctggatttcc
3421 tgaagtccga cggcttcgcc aacagaaact tcatgcagct gatccacgac gacagcctga
3481 cctttaaaga ggacatccag aaagcccagg tgtccggcca gggcgatagc ctgcacgagc
3541 acattgccaa tctggccggc agccccgcca ttaagaaggg catcctgcag acagtgaagg
3601 tggtggacga gctcgtgaaa gtgatgggcc ggcacaagcc cgagaacatc gtgatcgaaa
3661 tggccagaga gaaccagacc acccagaagg gacagaagaa cagccgcgag agaatgaagc
3721 ggatcgaaga gggcatcaaa gagctgggca gccagatcct gaaagaacac cccgtggaaa
3781 acacccagct gcagaacgag aagctgtacc tgtactacct gcagaatggg cgggatatgt
3841 acgtggacca ggaactggac atcaaccggc tgtccgacta cgatgtggac catatcgtgc
3901 ctcagagctt tctgaaggac gactccatcg acaacaaggt gctgaccaga agcgacaaga
3961 accggggcaa gagcgacaac gtgccctccg aagaggtcgt gaagaagatg aagaactact
4021 ggcggcagct gctgaacgcc aagctgatta cccagagaaa gttcgacaat ctgaccaagg
4081 ccgagagagg cggcctgagc gaactggata aggccggctt catcaagaga cagctggtgg
4141 aaacccggca gatcacaaag cacgtggcac agatcctgga ctcccggatg aacactaagt
4201 acgacgagaa tgacaagctg atccgggaag tgaaagtgat caccctgaag tccaagctgg
4261 tgtccgattt ccggaaggat ttccagtttt acaaagtgcg cgagatcaac aactaccacc
4321 acgcccacga cgcctacctg aacgccgtcg tgggaaccgc cctgatcaaa aagtacccta
4381 agctggaaag cgagttcgtg tacggcgact acaaggtgta cgacgtgcgg aagatgatcg
4441 ccaagagcga gcaggaaatc ggcaaggcta ccgccaagta cttcttctac agcaacatca
4501 tgaacttttt caagaccgag attaccctgg ccaacggcga gatccggaag cggcctctga
4561 tcgagacaaa cggcgaaacc ggggagatcg tgtgggataa gggccgggat tttgccaccg
4621 tgcggaaagt gctgagcatg ccccaagtga atatcgtgaa aaagaccgag gtgcagacag
4681 gcggcttcag caaagagtct atcctgccca agaggaacag cgataagctg atcgccagaa
4741 agaaggactg ggaccctaag aagtacggcg gcttcgacag ccccaccgtg gcctattctg
4801 tgctggtggt ggccaaagtg gaaaagggca agtccaagaa actgaagagt gtgaaagagc
4861 tgctggggat caccatcatg gaaagaagca gcttcgagaa gaatcccatc gactttctgg
4921 aagccaaggg ctacaaagaa gtgaaaaagg acctgatcat caagctgcct aagtactccc
4981 tgttcgagct ggaaaacggc cggaagagaa tgctggcctc tgccggcgaa ctgcagaagg
5041 gaaacgaact ggccctgccc tccaaatatg tgaacttcct gtacctggcc agccactatg
5101 agaagctgaa gggctccccc gaggataatg agcagaaaca gctgtttgtg gaacagcaca
5161 agcactacct ggacgagatc atcgagcaga tcagcgagtt ctccaagaga gtgatcctgg
5221 ccgacgctaa tctggacaaa gtgctgtccg cctacaacaa gcaccgggat aagcccatca
5281 gagagcaggc cgagaatatc atccacctgt ttaccctgac caatctggga gcccctgccg
5341 ccttcaagta ctttgacacc accatcgacc ggaagaggta caccagcacc aaagaggtgc
5401 tggacgccac cctgatccac cagagcatca ccggcctgta cgagacacgg atcgacctgt
5461 ctcagctggg aggcgacaaa aggccggcgg ccacgaaaaa ggccggccag gcaaaaaaga
5521 aaaagcttga gggcagagga agtctgctaa catgcggtga cgtggaggag aatcccggcc
5581 ctgctagcat ggtgagcaag ggcgaggagg ataacatggc catcatcaag gagttcatgc
5641 gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc gagggcgagg
5701 gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc aagggtggcc
5761 ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc aaggcctacg
5821 tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag ggcttcaagt
5881 gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag gactcctccc
5941 tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc ccctcagacg
6001 gccccgtaat gcagaagaaa accatgggct gggaggcctc ctccgagcgg atgtaccccg
6061 aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac ggcggccact
6121 acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg cccggcgcct
6181 acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc atcgtggaac
6241 agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg tacaagtaag
6301 aattcctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
6361 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
6421 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
6481 ggtggggcag gacagcaagg gggaggattg ggaagagaat agcaggcatg ctggggagcg
6541 gccgcaggaa cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac
6601 tgaggccggg cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag
6661 cgagcgagcg cgcagctgcc tgcaggggcg cctgatgcgg tattttctcc ttacgcatct
6721 gtgcggtatt tcacaccgca tacgtcaaag caaccatagt acgcgccctg tagcggcgca
6781 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
6841 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
6901 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
6961 cccaaaaaac ttgatttggg tgatggttca cgtagtgggc catcgccctg atagacggtt
7021 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
7081 acaacactca accctatctc gggctattct tttgatttat aagggatttt gccgatttcg
7141 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
7201 ttaacgttta caattttatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
7261 agccagcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
7321 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
7381 ccgtcatcac cgaaacgcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
7441 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
7501 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
7561 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
7621 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
7681 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
7741 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
7801 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
7861 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
7921 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
7981 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
8041 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
8101 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
8161 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
8221 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
8281 tggtttattg ctgataaatc tggagccggt gagcgtggaa gccgcggtat cattgcagca
8341 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
8401 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
8461 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
8521 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
8581 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
8641 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
8701 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
8761 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
8821 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
8881 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
8941 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
9001 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
9061 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
9121 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
9181 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
9241 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgt
//
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