| 质粒名称: | pdCas9-Tet1-CD |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 83340 |
| 存储实验室: | Ronggui Hu |
|
载体骨架基本信息 |
|
| 载体名称: | pX330 |
| 载体出品公司: | Received from Feng Zhang (MIT) |
| 载体抗性: | 氨苄霉素 |
| 载体类型: | 哺乳细胞表达，CRISPR |
| 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: | dCas9-Tet1-CD and sgRNA scaffold with 2xMS2 binding sites |
| 物种来源: | Synthetic |
| 基因突变: | D10A, H840A |
| 启动子 | U6, CBH |
| 克隆方法: | Gibson Cloning |
| 5' 测序引物 | U6 |
| 3' 测序引物 | BGH-rev |
|
文献引用方法 |
|
| 材料和方法部分: | pdCas9-Tet1-CD was a gift from Ronggui Hu (爱迪基因 plasmid # 83340) |
| 参考文献部分: | A CRISPR-based approach for targeted DNA demethylation. Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu R. Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9. eCollection 2016. 10.1038/celldisc.2016.9 PubMed 27462456 |
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 pdCas9-Tet1-CD 10468 bp ds-DNA circular SYN 10-JAN-2018
DEFINITION To introduce dCas9-fused Tet1-CD and sgRNA2.0 system.
ACCESSION .
VERSION .
KEYWORDS pdCas9-Tet1-CD
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10468)
AUTHORS Xu X, Tao Y, Gao X, Zhang L, Li X, Zou W, Ruan K, Wang F, Xu GL, Hu
R
TITLE A CRISPR-based approach for targeted DNA demethylation.
JOURNAL Cell Discov. 2016 May 3;2:16009. doi: 10.1038/celldisc.2016.9.
eCollection 2016.
PUBMED 27462456
COMMENT This file is created by Vector NTI
http://www.biofeng.com/
COMMENT ORIGDB|GenBank
COMMENT VNTAUTHORNAME|biofeng.com|
COMMENT VNTNAME|pdCas9-Tet1-CD|
FEATURES Location/Qualifiers
source 1..10468
/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 285..303
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 355..373
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
enhancer 416..701
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer;
contains an 18-bp deletion relative to the standard CMV
enhancer"
promoter 703..980
/label=chicken beta-actin promoter
intron 981..1209
/label=hybrid intron
/note="hybrid between chicken beta-actin (CBA) and minute
virus of mice (MMV) introns (Gray et al., 2011)"
CDS 1230..1256
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 1260..1280
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1290..5390
/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 H840A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5394..5414
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(7509..7526)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7515..7722
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
repeat_region 7731..7871
/label=AAV2 ITR
/note="inverted terminal repeat of adeno-associated virus
serotype 2"
repeat_region 7731..7860
/label=AAV2 ITR
rep_origin 7946..8401
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8033..8052)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8243..8264
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(8418..8437)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 8537..8559
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(8597..8615)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 8683..8787
/gene="bla"
/label=AmpR promoter
CDS 8788..9648
/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(9006..9025)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9819..10407
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10308..10327
/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 gccaacatga ggatcaccca
301 tgtctgcagg gcctagcaag ttaaaataag gctagtccgt tatcaacttg gccaacatga
361 ggatcaccca tgtctgcagg gccaagtggc accgagtcgg tgcttttttg gtacccgtta
421 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
481 caatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
541 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
601 atgacggtaa atggcccgcc tggcattgtg cccagtacat gaccttatgg gactttccta
661 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
721 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
781 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg
841 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
901 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
961 agcgaagcgc gcggcgggcg ggagtcgctg cgacgctgcc ttcgccccgt gccccgctcc
1021 gccgccgcct cgcgccgccc gccccggctc tgactgaccg cgttactccc acaggtgagc
1081 gggcgggacg gcccttctcc tccgggctgt aattagctga gcaagaggta agggtttaag
1141 ggatggttgg ttggtggggt attaatgttt aattacctgg agcacctgcc tgaaatcact
1201 ttttttcagg ttggaccggt gccaccatgt acccatacga tgttccagat tacgcttcgc
1261 cgaagaaaaa gcgcaaggtc gaagcgtccg acaagaagta cagcatcggc ctggccatcg
1321 gcaccaactc tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat
1381 tcaaggtgct gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc
1441 tgttcgacag cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat
1501 acaccagacg gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca
1561 aggtggacga cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga
1621 agcacgagcg gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt
1681 accccaccat ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc
1741 ggctgatcta tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg
1801 gcgacctgaa ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct
1861 acaaccagct gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc
1921 tgtctgccag actgagcaag agcagacggc tggaaaatct gatcgcccag ctgcccggcg
1981 agaagaagaa tggcctgttc ggcaacctga ttgccctgag cctgggcctg acccccaact
2041 tcaagagcaa cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg
2101 acgacgacct ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg
2161 ccgccaagaa cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga
2221 tcaccaaggc ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc
2281 tgaccctgct gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct
2341 tcgaccagag caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt
2401 tctacaagtt catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga
2461 agctgaacag agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc
2521 accagatcca cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat
2581 tcctgaagga caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg
2641 tgggccctct ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa
2701 ccatcacccc ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca
2761 tcgagcggat gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca
2821 gcctgctgta cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg
2881 agggaatgag aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc
2941 tgttcaagac caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa
3001 tcgagtgctt cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg
3061 gcacatacca cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa
3121 acgaggacat tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga
3181 tcgaggaacg gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga
3241 agcggcggag atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg
3301 acaagcagtc cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa
3361 acttcatgca gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc
3421 aggtgtccgg ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg
3481 ccattaagaa gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg
3541 gccggcacaa gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga
3601 agggacagaa gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg
3661 gcagccagat cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt
3721 acctgtacta cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc
3781 ggctgtccga ctacgatgtg gacgctatcg tgcctcagag ctttctgaag gacgactcca
3841 tcgacaacaa ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct
3901 ccgaagaggt cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga
3961 ttacccagag aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg
4021 ataaggccgg cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg
4081 cacagatcct ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg
4141 aagtgaaagt gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt
4201 tttacaaagt gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg
4261 tcgtgggaac cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg
4321 actacaaggt gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg
4381 ctaccgccaa gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc
4441 tggccaacgg cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga
4501 tcgtgtggga taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag
4561 tgaatatcgt gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc
4621 ccaagaggaa cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg
4681 gcggcttcga cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg
4741 gcaagtccaa gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa
4801 gcagcttcga gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa
4861 aggacctgat catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga
4921 gaatgctggc ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat
4981 atgtgaactt cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata
5041 atgagcagaa acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc
5101 agatcagcga gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt
5161 ccgcctacaa caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc
5221 tgtttaccct gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg
5281 accggaagag gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca
5341 tcaccggcct gtacgagaca cggatcgacc tgtctcagct gggaggcgac agccccaaga
5401 agaagagaaa ggtggaggcc agcggtggag gaagtggagg tggttcaggt ggaggatccg
5461 gcgaagctgc accctgtgac tgtgatggag gtacacaaaa agaaaaaggc ccatattata
5521 cacaccttgg ggcaggacca agtgtggctg ctgtcaggga gctcatggag actaggtttg
5581 gccagaaggg gaaggcaatc cggattgaga agatagtgtt cacggggaag gaagggaaga
5641 gctctcaggg ctgcccggtc gccaagtggg tgatcagaag aagtggtcct gaagagaagc
5701 ttatttgttt ggttcgtgag cgtgtagacc atcactgttc gacggctgtg atagttgtcc
5761 ttatcctgct gtgggaaggt atccctcgcc tgatggctga ccgcctgtac aaagagctca
5821 ctgagaactt gaggtcctac agcggacatc ccacagaccg aagatgtacc ctcaacaaaa
5881 agcgtacctg cacctgtcaa ggcatcgacc caaaaacctg cggagcgtcc ttctcctttg
5941 gctgttcgtg gagcatgtat ttcaacggct gtaagtttgg gaggagtgaa aaccccagaa
6001 aattcagact tgctccaaac taccccttac ataactacta taagagaatt actggaatga
6061 gttctgaagg aagtgacgtg aaaaccgggt ggatcattcc agaccgcaag accctcataa
6121 gcagagagga aaaacagctt gaaaagaatt tacaagaatt ggctacagta ttagctccac
6181 tttacaagca gatggctcca gttgcttatc aaaatcaggt ggaatatgaa gaagttgctg
6241 gagactgtcg acttggaaat gaagaggggc gtcctttctc tggtgtcacc tgttgcatgg
6301 atttttgtgc ccattctcac aaggacattc acaacatgca caacggaagc accgtggtgt
6361 gtacgttgat tcgagcagat ggccgtgaca caaattgtcc cgaggatgaa caactccacg
6421 tcctgccact ataccggctt gcagacactg atgaatttgg ctccgtggaa gggatgaagg
6481 ccaaaatcaa atctggggcc atccaagtca atgggccaac caggaagagg cgactacgtt
6541 ttactgagcc tgttcctcga tgtgggaaga gggccaaaat gaagcagaac cacaataaat
6601 caggttcaca caacactaag agcttttcat cagcctcatc tacttctcac ctagtgaaag
6661 acgaatctac agacttctgt cccctgcagg cttcctccgc agaaacatct acctgtacgt
6721 acagtaaaac agcctcaggt gggtttgcag aaacaagtag tattctccac tgcacaatgc
6781 cttctggagc acacagtggt gctaatgcag ctgctgggga atgtactgga acggtgcagc
6841 ctgccgaggt ggctgctcat cctcaccagt ctcttcccac agccgattct cccgttcatg
6901 ctgagcctct cactagtcca tctgagcagc taacttctaa ccagtcaaac cagcagctcc
6961 ctctcctcag caattctcag aaactggctt cctgtcaggt ggaagatgag cggcaccctg
7021 aagcggatga gcctcagcac cccgaggacg ataacttgcc tcaacttgat gagttctggt
7081 cagacagtga ggagatctac gccgatcctt cctttggtgg cgtggcgata gcacccattc
7141 acggctcggt gctcattgag tgcgctcgga aggagcttca tgctaccacc tctttgcgct
7201 cccccaaacg aggggtccct tttcgtgtgt cccttgtatt ctaccagcac aaaagcctaa
7261 acaagcctaa tcatggtttt gatatcaaca aaattaagtg taaatgcaaa aaagtaacga
7321 aaaaaaagcc cgcagaccgg gagtgtcctg atgtatcccc cgaagccaat ttatcacacc
7381 aaattccttc tcgagttgca tcaaccttaa cccgagacaa tgttgttacc gtgtccccat
7441 actctctcac tcatgttgcg ggaccctaca atcgttgggt ctaagaattc ctagagctcg
7501 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
7561 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
7621 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
7681 caagggggag gattgggaag agaatagcag gcatgctggg gagcggccgc aggaacccct
7741 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc
7801 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag
7861 ctgcctgcag gggcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca
7921 ccgcatacgt caaagcaacc atagtacgcg ccctgtagcg gcgcattaag cgcggcgggt
7981 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
8041 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
8101 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
8161 ttgggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
8221 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
8281 atctcgggct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
8341 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gtttacaatt
8401 ttatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac
8461 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
8521 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
8581 cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata
8641 atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
8701 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
8761 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
8821 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
8881 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
8941 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
9001 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
9061 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
9121 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
9181 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
9241 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
9301 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
9361 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
9421 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
9481 aaatctggag ccggtgagcg tggaagccgc ggtatcattg cagcactggg gccagatggt
9541 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
9601 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
9661 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
9721 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
9781 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
9841 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
9901 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
9961 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
10021 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
10081 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
10141 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
10201 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
10261 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
10321 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
10381 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
10441 gccttttgct ggccttttgc tcacatgt
//
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