| 质粒名称: | p2T-CMV-miniSdd7-BE4max-BlastR |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 204851 |
| 存储实验室: | Caixia Gao |
|
载体骨架基本信息 |
|
| 载体名称: | p2T-CMV-ABEmax-BlastR |
| 载体出品公司: | David Liu |
| 完整质粒大小: | 10259bp |
| 载体类型: | 哺乳细胞表达, CRISPR |
| 筛选标记: | Blasticidin |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | miniSdd7-nSpCas9(D10A)-UGI |
| 物种来源: | Synthetic |
| 基因突变: | D10A in SpCas9 |
| 启动子: | CMV, SV40, EM7 |
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: |
p2T-CMV-miniSdd7-BE4max-BlastR was a gift from Caixia Gao (Addgene plasmid # 204851 ; http://n2t.net/addgene:204851 ; RRID:Addgene_204851)
|
| 参考文献部分: | Discovery of deaminase functions by structure-based protein clustering. Huang J, Lin Q, Fei H, He Z, Xu H, Li Y, Qu K, Han P, Gao Q, Li B, Liu G, Zhang L, Hu J, Zhang R, Zuo E, Luo Y, Ran Y, Qiu JL, Zhao KT, Gao C. Cell. 2023 Jul 20;186(15):3182-3195.e14. doi: 10.1016/j.cell.2023.05.041. Epub 2023 Jun 27. 10.1016/j.cell.2023.05.041 PubMed 37379837 |
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 p2T-CMV-miniSdd7-BE4max-BlastR 10259 bp ds-DNA circular SYN 17-AUG-2023
DEFINITION For cytosine base editing using miniSdd7 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS p2T-CMV-miniSdd7-BE4max-BlastR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10259)
AUTHORS Huang J, Lin Q, Fei H, He Z, Xu H, Li Y, Qu K, Han P, Gao Q, Li B,
Liu B, Zhang L, Hu J, Zhang R, Zuo E, Luo Y, Ran Y, Qiu J-L, Zhao
KT, Gao C
TITLE Discovery of deaminase functions by structure-based protein
clustering
JOURNAL Cell 2023
REFERENCE 2 (bases 1 to 10259)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 17, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10259
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 1..204
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 155..175
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 246..265
/label=T7
/note="T7 promoter, forward primer"
promoter 246..264
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 314..334
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 902..5002
/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 5033..5281
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5312..5560
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5603..5623
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5632..5649
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(5672..5689)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 5678..5902
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(5973..5989)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(5973..5989)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(5986..6008)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 5997..6013
/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)."
promoter complement(6021..6051)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6066..6087
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
polyA_signal 6204..6325
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(6215..6234)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 6269..6288
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(6483..6881)
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
promoter complement(6900..6947)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(6995..7324)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7009..7144
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(7063..7082)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 7309..7329
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(7708..8296)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7788..7807)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8467..9327)
/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 9090..9109
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9328..9432)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9511..9530)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 9880..10259
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
ORIGIN
1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gagatccgcg
241 gccgctaata cgactcacta tagggagagc cgccaccatg aaacggacag ccgacggaag
301 cgagttcgag tcaccaaaga agaagcggaa agtcgaagga ggtggccctg gggcagttcc
361 tgagggtggc gacggcccac ctgccgtccc tgccgaggag gtggaaagac tgcgcggaga
421 gctgcctccc cctgtggtgc cagggactgg gcaaaagacc cacggcagat ggatcggccc
481 tgacggtaga gtccgggcca tcgtgagcgg aagagatgaa gatgctgcac tggtgcacgc
541 ccagctggct gccaagggca tcccagacga gcccaccagg aactctgatg tggagcagaa
601 gctggcagcc cacatggtgg cgaatgggat caggcatgtg accctggtta tcaaccacag
661 gccttgcaga ggcttcgatg actcctgtga taccctggtg cctatcattc tgccagaagg
721 ctgcacactg accgtgcatg gccagacaga caaaggcatg agagtccgag tgcgctacac
781 aggaggagcc cggccctggt ggagctccgg aggatctagc ggaggctcct ctggctctga
841 gacacctggc acaagcgaga gcgcaacacc tgaaagcagc gggggcagca gcggggggtc
901 agacaagaag tacagcatcg gcctggccat cggcaccaac tctgtgggct gggccgtgat
961 caccgacgag tacaaggtgc ccagcaagaa attcaaggtg ctgggcaaca ccgaccggca
1021 cagcatcaag aagaacctga tcggagccct gctgttcgac agcggcgaaa cagccgaggc
1081 cacccggctg aagagaaccg ccagaagaag atacaccaga cggaagaacc ggatctgcta
1141 tctgcaagag atcttcagca acgagatggc caaggtggac gacagcttct tccacagact
1201 ggaagagtcc ttcctggtgg aagaggataa gaagcacgag cggcacccca tcttcggcaa
1261 catcgtggac gaggtggcct accacgagaa gtaccccacc atctaccacc tgagaaagaa
1321 actggtggac agcaccgaca aggccgacct gcggctgatc tatctggccc tggcccacat
1381 gatcaagttc cggggccact tcctgatcga gggcgacctg aaccccgaca acagcgacgt
1441 ggacaagctg ttcatccagc tggtgcagac ctacaaccag ctgttcgagg aaaaccccat
1501 caacgccagc ggcgtggacg ccaaggccat cctgtctgcc agactgagca agagcagacg
1561 gctggaaaat ctgatcgccc agctgcccgg cgagaagaag aatggcctgt tcggaaacct
1621 gattgccctg agcctgggcc tgacccccaa cttcaagagc aacttcgacc tggccgagga
1681 tgccaaactg cagctgagca aggacaccta cgacgacgac ctggacaacc tgctggccca
1741 gatcggcgac cagtacgccg acctgtttct ggccgccaag aacctgtccg acgccatcct
1801 gctgagcgac atcctgagag tgaacaccga gatcaccaag gcccccctga gcgcctctat
1861 gatcaagaga tacgacgagc accaccagga cctgaccctg ctgaaagctc tcgtgcggca
1921 gcagctgcct gagaagtaca aagagatttt cttcgaccag agcaagaacg gctacgccgg
1981 ctacattgac ggcggagcca gccaggaaga gttctacaag ttcatcaagc ccatcctgga
2041 aaagatggac ggcaccgagg aactgctcgt gaagctgaac agagaggacc tgctgcggaa
2101 gcagcggacc ttcgacaacg gcagcatccc ccaccagatc cacctgggag agctgcacgc
2161 cattctgcgg cggcaggaag atttttaccc attcctgaag gacaaccggg aaaagatcga
2221 gaagatcctg accttccgca tcccctacta cgtgggccct ctggccaggg gaaacagcag
2281 attcgcctgg atgaccagaa agagcgagga aaccatcacc ccctggaact tcgaggaagt
2341 ggtggacaag ggcgcttccg cccagagctt catcgagcgg atgaccaact tcgataagaa
2401 cctgcccaac gagaaggtgc tgcccaagca cagcctgctg tacgagtact tcaccgtgta
2461 taacgagctg accaaagtga aatacgtgac cgagggaatg agaaagcccg ccttcctgag
2521 cggcgagcag aaaaaggcca tcgtggacct gctgttcaag accaaccgga aagtgaccgt
2581 gaagcagctg aaagaggact acttcaagaa aatcgagtgc ttcgactccg tggaaatctc
2641 cggcgtggaa gatcggttca acgcctccct gggcacatac cacgatctgc tgaaaattat
2701 caaggacaag gacttcctgg acaatgagga aaacgaggac attctggaag atatcgtgct
2761 gaccctgaca ctgtttgagg acagagagat gatcgaggaa cggctgaaaa cctatgccca
2821 cctgttcgac gacaaagtga tgaagcagct gaagcggcgg agatacaccg gctggggcag
2881 gctgagccgg aagctgatca acggcatccg ggacaagcag tccggcaaga caatcctgga
2941 tttcctgaag tccgacggct tcgccaacag aaacttcatg cagctgatcc acgacgacag
3001 cctgaccttt aaagaggaca tccagaaagc ccaggtgtcc ggccagggcg atagcctgca
3061 cgagcacatt gccaatctgg ccggcagccc cgccattaag aagggcatcc tgcagacagt
3121 gaaggtggtg gacgagctcg tgaaagtgat gggccggcac aagcccgaga acatcgtgat
3181 cgaaatggcc agagagaacc agaccaccca gaagggacag aagaacagcc gcgagagaat
3241 gaagcggatc gaagagggca tcaaagagct gggcagccag atcctgaaag aacaccccgt
3301 ggaaaacacc cagctgcaga acgagaagct gtacctgtac tacctgcaga atgggcggga
3361 tatgtacgtg gaccaggaac tggacatcaa ccggctgtcc gactacgatg tggaccatat
3421 cgtgcctcag agctttctga aggacgactc catcgacaac aaggtgctga ccagaagcga
3481 caagaaccgg ggcaagagcg acaacgtgcc ctccgaagag gtcgtgaaga agatgaagaa
3541 ctactggcgg cagctgctga acgccaagct gattacccag agaaagttcg acaatctgac
3601 caaggccgag agaggcggcc tgagcgaact ggataaggcc ggcttcatca agagacagct
3661 ggtggaaacc cggcagatca caaagcacgt ggcacagatc ctggactccc ggatgaacac
3721 taagtacgac gagaatgaca agctgatccg ggaagtgaaa gtaatcaccc tgaagtccaa
3781 gctggtgtcc gatttccgga aggatttcca gttttacaaa gtgcgcgaga tcaacaacta
3841 ccaccacgcc cacgacgcct acctgaacgc cgtcgtggga accgccctga tcaaaaagta
3901 ccctaagctg gaaagcgagt tcgtgtacgg cgactacaag gtgtacgacg tgcggaagat
3961 gatcgccaag agcgagcagg aaatcggcaa ggctaccgcc aagtacttct tctacagcaa
4021 catcatgaac tttttcaaga ccgagattac cctggccaac ggcgagatcc ggaagcggcc
4081 tctaatcgag acaaacggcg aaaccgggga gatcgtgtgg gataagggcc gggattttgc
4141 caccgtgcgg aaagtgctga gcatgcccca agtgaatatc gtgaaaaaga ccgaggtgca
4201 gacaggcggc ttcagcaaag agtctatcct gcccaagagg aacagcgata agctgatcgc
4261 cagaaagaag gactgggacc ctaagaagta cggcggcttc gacagcccca ccgtggccta
4321 ttctgtgctg gtggtggcca aagtggaaaa gggcaagtcc aagaaactga agagtgtgaa
4381 agagctgctg gggatcacca tcatggaaag aagcagcttc gagaagaatc ccatcgactt
4441 tctggaagcc aagggctaca aagaagtgaa aaaggacctg atcatcaagc tgcctaagta
4501 ctccctgttc gagctggaaa acggccggaa gagaatgctg gcctctgccg gcgaactgca
4561 gaagggaaac gaactggccc tgccctccaa atatgtgaac ttcctgtacc tggccagcca
4621 ctatgagaag ctgaagggct cccccgagga taatgagcag aaacagctgt ttgtggaaca
4681 gcacaagcac tacctggacg agatcatcga gcagatcagc gagttctcca agagagtgat
4741 cctggccgac gctaatctgg acaaagtgct gtccgcctac aacaagcacc gggataagcc
4801 catcagagag caggccgaga atatcatcca cctgtttacc ctgaccaatc tgggagcccc
4861 tgccgccttc aagtactttg acaccaccat cgaccggaag aggtacacca gcaccaaaga
4921 ggtgctggac gccaccctga tccaccagag catcaccggc ctgtacgaga cacggatcga
4981 cctgtctcag ctgggaggtg acagcggcgg gagcggcggg agcgggggga gcactaatct
5041 gagcgacatc attgagaagg agactgggaa acagctggtc attcaggagt ccatcctgat
5101 gctgcctgag gaggtggagg aagtgatcgg caacaagcca gagtctgaca tcctggtgca
5161 caccgcctac gacgagtcca cagatgagaa tgtgatgctg ctgacctctg acgcccccga
5221 gtataagcct tgggccctgg tcatccagga ttctaacggc gagaataaga tcaagatgct
5281 gagcggagga tccggaggat ctggaggcag caccaacctg tctgacatca tcgagaagga
5341 gacaggcaag cagctggtca tccaggagag catcctgatg ctgcccgaag aagtcgaaga
5401 agtgatcgga aacaagcctg agagcgatat cctggtccat accgcctacg acgagagtac
5461 cgacgaaaat gtgatgctgc tgacatccga cgccccagag tataagccct gggctctggt
5521 catccaggat tccaacggag agaacaaaat caaaatgctg tctggcggct caaaaagaac
5581 cgccgacggc agcgaattcg agcccaagaa gaagaggaaa gtctaaccgg tcatcatcac
5641 catcaccatt gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
5701 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
5761 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
5821 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
5881 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctcgataccg
5941 tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
6001 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctagggt
6061 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
6121 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg
6181 cgtattgggc gctacagaca tgataagata cattgatgag tttggacaaa ccacaactag
6241 aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac
6301 cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc
6361 gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga
6421 aggcggcggt ggaatcgaaa tctcgtagca cgtgtcagtc ctgctcctcg gccacgaagt
6481 gcttagccct cccacacata accagagggc agcaattcac gaatcccaac tgccgtcggc
6541 tgtccatcac tgtccttcac tatggctttg atcccaggat gcagatcgag aagcacctgt
6601 cggcaccgtc cgcaggggct caagatgccc ctgttctcat ttccgatcgc gacgatacaa
6661 gtcaggttgc cagctgccgc agcagcagca gtgcccagca ccacgagttc tgcacaaggt
6721 cccccagtaa aatgatatac attgacacca gtaaagatgc ggccgtcgct agagagagct
6781 gcgctggcga cgctgtagtc ttcagagatg gggatgctgt tgattgtagc cgttgctctt
6841 tcaatgaggg tggattcttc ttgagacaaa ggcttggcca tggtttagtt cctcaccttg
6901 tcgtattata ctatgccgat atactatgcc gatgattaat tgtcaacacg tgctgatcag
6961 atccgaaaat ggatatacaa gctcccggga gctttttgca aaagcctagg cctccaaaaa
7021 agcctcctca ctacttctgg aatagctcag aggcaaaggc ggcctcggcc tctgcataaa
7081 taaaaaaaat tagtcagcca tggggcggag aatgggcgga actgggcgga gttaggggcg
7141 ggatgggcgg agttaggggc gggactatgg ttgctgacta attgagatgc atgctttgca
7201 tacttctgcc tgctggggag cctggggact ttccacacct ggttgctgac taattgagat
7261 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac cctaactgac
7321 acacattcca cagaattcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
7381 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
7441 aacatgtaag tgatctccaa aaaataagta ctttttgact gtaaataaaa ttgtaaggag
7501 taaaaagtac ttttttttct aaaaaaatgt aattaagtaa aagtaaaagt attgattttt
7561 aattgtactc aagtaaagta aaaatcccca aaaataatac ttaagtacag taatcaagta
7621 aaattactca agtactttac acctctgcat gtgagcaaaa ggccagcaaa aggccaggaa
7681 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
7741 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
7801 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
7861 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
7921 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
7981 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
8041 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
8101 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
8161 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
8221 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
8281 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgaca ctcagtggaa
8341 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
8401 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
8461 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
8521 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
8581 tggccccagt gctgcaatga taccgcggga cccacgctca ccggctccag atttatcagc
8641 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
8701 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
8761 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
8821 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
8881 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
8941 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
9001 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
9061 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
9121 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
9181 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
9241 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
9301 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
9361 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
9421 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatcg
9481 atctcccgat cccctagggt cgactctcag tacaatctgc tctgatgccg catagttaag
9541 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta
9601 agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg
9661 ttttgcgctg cttcgcgatg tacgggccag atatacgcgg gtaccggcat atggttcttg
9721 acagaggtgt aaaaagtact caaaaatttt actcaagtga aagtacaagt acttagggaa
9781 aattttactc aattaaaagt aaaagtatct ggctagaatc ttacttgagt aaaagtaaaa
9841 aagtactcca ttaaaattgt acttgagtat taacgcgttg acattgatta ttgactagtt
9901 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
9961 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
10021 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
10081 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
10141 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
10201 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatg
//
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