| 质粒名称: | ER50-SpCas9-ER50 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 85448 |
| 存储实验室: | Amit Choudhary |
|
载体骨架基本信息 |
|
| 载体名称: | px330 |
| 载体出品公司: | Feng Zhang |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 4234 bp |
| 完整质粒大小: | 10000 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达.AAV |
| 筛选标记: | -- |
| 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: | ER50-SpCas9-ER50,ESpE |
| 物种来源: | -- |
| 插入大小: | 5766 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | Cbh |
| 标签/融合蛋白: |
雌激素受体配体结合域的不稳定结构域（插入端N端）
雌激素受体配体结合结构域的不稳定结构域（插入C端）
|
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | agcgaagcgcgcggcgggcg |
| 3' 测序引物 | TAGAAGGCACAGTCGAGG |
|
文献引用方法 |
|
| 材料和方法部分: | ER50-SpCas9-ER50 was a gift from Amit Choudhary (Addgene plasmid # 85448 ; http://n2t.net/addgene:85448 ; RRID:Addgene_85448) |
| 参考文献部分: | Multidimensional chemical control of CRISPR-Cas9. Maji B, Moore CL, Zetsche B, Volz SE, Zhang F, Shoulders MD, Choudhary A. Nat Chem Biol. 2017 Jan;13(1):9-11. doi: 10.1038/nchembio.2224. Epub 2016 Oct 31. 10.1038/nchembio.2224 PubMed 27820801 |
买家导航
ER50-SpCas9-ER50载体质粒基本信息
ER50-SpCas9-ER50质粒图谱载体图谱和ER50-SpCas9-ER50载体序列质粒序列多克隆位点信息
ER50-SpCas9-ER50质粒载体简介
ER50-SpCas9-ER50质粒序列载体序列
LOCUS ER50-SpCas9-ER50 10000 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses SpCas9 fused to ER50 domain on both N- and C-termini in
mammalian cells.
ACCESSION .
VERSION .
KEYWORDS ER50-SpCas9-ER50
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10000)
AUTHORS Maji B, Moore CL, Zetsche B, Volz SE, Zhang F, Shoulders MD,
Choudhary A
TITLE Multidimensional chemical control of CRISPR-Cas9.
JOURNAL Nat Chem Biol. 2017 Jan;13(1):9-11. doi: 10.1038/nchembio.2224. Epub
2016 Oct 31.
PUBMED 27820801
REFERENCE 2 (bases 1 to 10000)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10000
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..241
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 1..21
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 172..191
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 268..343
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
enhancer 440..725
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 727..1004
/label=chicken beta-actin promoter
intron 1005..1233
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
regulatory 1245..1254
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1254..1319
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 1326..1346
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2130..6230
/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 6231..6278
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
primer_bind complement(7041..7058)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7047..7254
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 7263..7403
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 7263..7392
/label=AAV2 ITR
rep_origin 7478..7933
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7565..7584)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7775..7796
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7950..7969)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8069..8091
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(8129..8147)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 8215..8319
/gene="bla"
/label=AmpR promoter
CDS 8320..9180
/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(8538..8557)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9351..9939
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 9840..9859
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gagggcctat ttcccatgat tccttcatat ttgcatatac gatacaaggc tgttagagag
61 ataattggaa ttaatttgac tgtaaacaca aagatattag tacaaaatac gtgacgtaga
121 aagtaataat ttcttgggta gtttgcagtt ttaaaattat gttttaaaat ggactatcat
181 atgcttaccg taacttgaaa gtatttcgat ttcttggctt tatatatctt gtggaaagga
241 cgaaacaccg ggtcttcgag aagacctgtt ttagagctag aaatagcaag ttaaaataag
301 gctagtccgt tatcaacttg aaaaagtggc accgagtcgg tgcttttttg ttttagagct
361 agaaatagca agttaaaata aggctagtcc gtttttagcg cgtgcgccaa ttctgcagac
421 aaatggctct agaggtaccc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
481 ccaacgaccc ccgcccattg acgtcaatag taacgccaat agggactttc cattgacgtc
541 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
601 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tgtgcccagt
661 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
721 ccatggtcga ggtgagcccc acgttctgct tcactctccc catctccccc ccctccccac
781 ccccaatttt gtatttattt attttttaat tattttgtgc agcgatgggg gcgggggggg
841 ggggggggcg cgcgccaggc ggggcggggc ggggcgaggg gcggggcggg gcgaggcgga
901 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
961 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgacgc
1021 tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc gcccgccccg gctctgactg
1081 accgcgttac tcccacaggt gagcgggcgg gacggccctt ctcctccggg ctgtaattag
1141 ctgagcaaga ggtaagggtt taagggatgg ttggttggtg gggtattaat gtttaattac
1201 ctggagcacc tgcctgaaat cacttttttt caggttggac cggtgccacc atggactata
1261 aggaccacga cggagactac aaggatcatg atattgatta caaagacgat gacgataaga
1321 tggccccaaa gaagaagcgg aaggtcggta tccacggagt cccagcagcc ggatccagcc
1381 ttgccctgtc acttacagcc gaccagatgg tttccgcgct tctcgacgct gaacctccaa
1441 ttctctattc cgaatacgac ccaaccaggc cgttctccga ggcatctatg atgggtctgc
1501 tgacaaatct ggcagacagg gaactggtgc acatgatcaa ttgggcgaag cgcgtacccg
1561 gattcgtcga tcttgcactc catgatcagg tgcacttgct ggagtgcgct tggatggaga
1621 tcctcatgat cgggctggtg tggcggagta tggaacaccc cggcaagttg ctgtttgcgc
1681 ctaacctcct gttggacagg aaccagggga aatgtgtgga gggcggtgtg gaaatctttg
1741 acatgctcct cgctacctca agccggttta ggatgatgaa tctgcagggc gaagagttcg
1801 tgtgtctcaa atctatcata ctgttgaaca gcggagtcta caccttcctc tccagtactc
1861 tgaaatctct ggaggagaaa gatcatatcc atcgcgtgct ggacaagata accgacacgt
1921 tgattcactt gatggccaaa gctgggctca ctctgcaaca acaacatcag cgactggcac
1981 agctgttgct gattttgagc cacattcggc acatgtccag caagagaatg gagcacctct
2041 atagtatgaa gtgcaagaac gtcgtacccc tgtcagatct gcttcttgaa atgcttgatg
2101 cccaccggct gggaagcgga agcggatccg acaagaagta cagcatcggc ctggacatcg
2161 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
2221 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
2281 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
2341 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
2401 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
2461 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
2521 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
2581 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
2641 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
2701 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
2761 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
2821 agaagaagaa tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact
2881 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
2941 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
3001 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
3061 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
3121 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
3181 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
3241 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
3301 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
3361 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
3421 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
3481 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
3541 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
3601 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
3661 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
3721 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
3781 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
3841 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
3901 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
3961 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
4021 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
4081 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
4141 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
4201 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
4261 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
4321 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
4381 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
4441 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
4501 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
4561 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
4621 ggctgtccga ctacgatgtg gaccatatcg tgcctcagag ctttctgaag gacgactcca
4681 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
4741 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
4801 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
4861 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
4921 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
4981 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
5041 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
5101 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
5161 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
5221 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
5281 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
5341 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
5401 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
5461 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
5521 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
5581 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
5641 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
5701 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
5761 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
5821 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
5881 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
5941 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
6001 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
6061 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
6121 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
6181 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac aaaaggccgg
6241 cggccacgaa aaaggccggc caggcaaaaa agaaaaagag ccttgccctg tcacttacag
6301 ccgaccagat ggtttccgcg cttctcgacg ctgaacctcc aattctctat tccgaatacg
6361 acccaaccag gccgttctcc gaggcatcta tgatgggtct gctgacaaat ctggcagaca
6421 gggaactggt gcacatgatc aattgggcga agcgcgtacc cggattcgtc gatcttgcac
6481 tccatgatca ggtgcacttg ctggagtgcg cttggatgga gatcctcatg atcgggctgg
6541 tgtggcggag tatggaacac cccggcaagt tgctgtttgc gcctaacctc ctgttggaca
6601 ggaaccaggg gaaatgtgtg gagggcggtg tggaaatctt tgacatgctc ctcgctacct
6661 caagccggtt taggatgatg aatctgcagg gcgaagagtt cgtgtgtctc aaatctatca
6721 tactgttgaa cagcggagtc tacaccttcc tctccagtac tctgaaatct ctggaggaga
6781 aagatcatat ccatcgcgtg ctggacaaga taaccgacac gttgattcac ttgatggcca
6841 aagctgggct cactctgcaa caacaacatc agcgactggc acagctgttg ctgattttga
6901 gccacattcg gcacatgtcc agcaagagaa tggagcacct ctatagtatg aagtgcaaga
6961 acgtcgtacc cctgtcagat ctgcttcttg aaatgcttga tgcccaccgg ctgtaagaat
7021 tcctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg
7081 cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata
7141 aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt
7201 ggggcaggac agcaaggggg aggattggga agagaatagc aggcatgctg gggagcggcc
7261 gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga
7321 ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga
7381 gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg
7441 cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta
7501 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
7561 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
7621 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
7681 aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt
7741 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
7801 acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggcc
7861 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
7921 acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
7981 cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca
8041 tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg
8101 tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat
8161 gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga
8221 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
8281 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
8341 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
8401 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
8461 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
8521 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
8581 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
8641 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
8701 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
8761 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
8821 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
8881 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
8941 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
9001 tttattgctg ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg
9061 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
9121 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
9181 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
9241 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
9301 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
9361 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
9421 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
9481 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
9541 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
9601 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
9661 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
9721 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
9781 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
9841 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
9901 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
9961 ttacggttcc tggccttttg ctggcctttt gctcacatgt
//
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