| 质粒名称: | pCMV-Target-ACEmax |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 139105 |
| 存储实验室: | Nozomu Yachie |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体类型: | 哺乳动物表达,CRISPR |
| 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: | Bi_partite_NLS-TadA*-XTEN_linker-TadA-XTEN_linker-SpCas9(D10A)-SH3_linker-PmCDA1-UGI-SV40NLS |
| 物种来源: | Synthetic |
| 插入大小: | 6534 bp |
| 启动子: | CMV promoter |
| 克隆方法: | Gibson克隆 |
| 5’测序引物 | CGTGTACGGTGGGAGGTCTA |
| 3’测序引物 | CTGGCAACTAGAAGGCACAG |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-Target-ACEmax was a gift from Nozomu Yachie (Addgene plasmid # 139105 ; http://n2t.net/addgene:139105 ; RRID:Addgene_139105) |
| 参考文献部分: | Base editors for simultaneous introduction of C-to-T and A-to-G mutations. Sakata RC, Ishiguro S, Mori H, Tanaka M, Tatsuno K, Ueda H, Yamamoto S, Seki M, Masuyama N, Nishida K, Nishimasu H, Arakawa K, Kondo A, Nureki O, Tomita M, Aburatani H, Yachie N. Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0509-0. doi: 10.1038/s41587-020-0509-0. 10.1038/s41587-020-0509-0 PubMed 32483365 |
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 pCMV-Target-ACEmax 9913 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Mammalian Target-ACEmax expression vector (pSI-625).
ACCESSION .
VERSION .
KEYWORDS pCMV-Target-ACEmax
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9913)
AUTHORS Sakata RC, Ishiguro S, Mori H, Tanaka M, Tatsuno K, Ueda H, Yamamoto
S, Seki M, Masuyama N, Nishida K, Nishimasu H, Arakawa K, Kondo A,
Nureki O, Tomita M, Aburatani H, Yachie N
TITLE Base editors for simultaneous introduction of C-to-T and A-to-G
mutations.
JOURNAL Nat Biotechnol. 2020 Jun 1. pii: 10.1038/s41587-020-0509-0. doi:
10.1038/s41587-020-0509-0.
PUBMED 32483365
REFERENCE 2 (bases 1 to 9913)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9913
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 19..398
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 399..602
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 553..573
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 644..663
/label=T7
/note="T7 promoter, forward primer"
promoter 644..662
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 712..732
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 1921..6021
/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 6229..6294
/codon_start=1
/product="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/label=3xFLAG
/translation="DYKDHDGDYKDHDIDYKDDDDK"
CDS 6928..6948
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6955..7209
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="MTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHT
AYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
primer_bind complement(7235..7252)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7241..7465
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7536..7552)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7536..7552)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7549..7571)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7560..7576
/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(7584..7614)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7629..7650
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7767..7784)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7938..8526)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8018..8037)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8697..9557)
/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 9320..9339
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9558..9662)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9741..9760)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
61 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
121 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
181 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
241 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
301 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
361 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
421 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
481 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
541 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
601 ctggtttagt gaaccgtcag atccgctaga gatccgcggc cgctaatacg actcactata
661 gggagagccg ccaccatgaa acggacagcc gacggaagcg agttcgagtc accaaagaag
721 aagcggaaag tctctgaagt cgagtttagc cacgagtatt ggatgaggca cgcactgacc
781 ctggcaaagc gagcatggga tgaaagagaa gtccccgtgg gcgccgtgct ggtgcacaac
841 aatagagtga tcggagaggg atggaacagg ccaatcggcc gccacgaccc taccgcacac
901 gcagagatca tggcactgag gcagggaggc ctggtcatgc agaattaccg cctgatcgat
961 gccaccctgt atgtgacact ggagccatgc gtgatgtgcg caggagcaat gatccacagc
1021 aggatcggaa gagtggtgtt cggagcacgg gacgccaaga ccggcgcagc aggctccctg
1081 atggatgtgc tgcaccaccc cggcatgaac caccgggtgg agatcacaga gggaatcctg
1141 gcagacgagt gcgccgccct gctgagcgat ttctttagaa tgcggagaca ggagatcaag
1201 gcccagaaga aggcacagag ctccaccgac tctggaggat ctagcggagg atcctctgga
1261 agcgagacac caggcacaag cgagtccgcc acaccagaga gctccggcgg ctcctccgga
1321 ggatcctctg aggtggagtt ttcccacgag tactggatga gacatgccct gaccctggcc
1381 aagagggcac gcgatgagag ggaggtgcct gtgggagccg tgctggtgct gaacaataga
1441 gtgatcggcg agggctggaa cagagccatc ggcctgcacg acccaacagc ccatgccgaa
1501 attatggccc tgagacaggg cggcctggtc atgcagaact acagactgat tgacgccacc
1561 ctgtacgtga cattcgagcc ttgcgtgatg tgcgccggcg ccatgatcca ctctaggatc
1621 ggccgcgtgg tgtttggcgt gaggaacgca aaaaccggcg ccgcaggctc cctgatggac
1681 gtgctgcact accccggcat gaatcaccgc gtcgaaatta ccgagggaat cctggcagat
1741 gaatgtgccg ccctgctgtg ctatttcttt cggatgccta gacaggtgtt caatgctcag
1801 aagaaggccc agagctccac cgactccgga ggatctagcg gaggctcctc tggctctgag
1861 acacctggca caagcgagag cgcaacacct gaaagcagcg ggggcagcag cggggggtca
1921 gacaagaagt acagcatcgg cctggccatc ggcaccaact ctgtgggctg ggccgtgatc
1981 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
2041 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
2101 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
2161 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
2221 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
2281 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
2341 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
2401 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
2461 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
2521 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagacgg
2581 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
2641 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
2701 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
2761 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
2821 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
2881 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
2941 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
3001 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
3061 aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct gctgcggaag
3121 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
3181 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
3241 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
3301 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
3361 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
3421 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
3481 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
3541 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
3601 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
3661 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
3721 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
3781 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
3841 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
3901 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
3961 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
4021 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
4081 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
4141 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
4201 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
4261 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
4321 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
4381 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggaccatatc
4441 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
4501 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
4561 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
4621 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
4681 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
4741 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
4801 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
4861 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
4921 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
4981 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
5041 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
5101 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
5161 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
5221 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
5281 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
5341 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
5401 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
5461 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
5521 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
5581 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
5641 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
5701 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
5761 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
5821 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
5881 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
5941 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
6001 ctgtctcagc tgggaggtga cggcggagga ggaactggag gaggaggatc agcagaatat
6061 gtgagagcac tgtttgactt taatgggaat gacgaagaag acctgccctt caagaagggc
6121 gacatcctgc ggatcagaga taagcctgag gagcagtggt ggaacgccga ggactctgag
6181 ggcaagagag gcatgatcct ggtgccctac gtggagaagt acagcggcga ttataaggac
6241 cacgatggcg actacaagga ccacgacatc gactacaagg atgacgatga caagtctggc
6301 atgacagatg ccgagtatgt gagaatccac gagaagctgg acatctacac cttcaagaag
6361 cagtttttca acaataagaa gagcgtgtcc cacagatgct acgtgctgtt tgagctgaag
6421 agaaggggcg agcggagagc ctgtttttgg ggctacgccg tgaacaagcc ccagtctggc
6481 acagagcggg gcatccacgc cgagatcttc tccatccgga aggtggagga gtacctgcgg
6541 gacaacccag gccagttcac catcaactgg tacagctcct ggtccccatg tgccgactgt
6601 gccgagaaga tcctggagtg gtacaaccag gagctgcggg gcaacggcca caccctgaag
6661 atctgggcct gcaagctgta ctatgagaag aatgccagaa accagatcgg cctgtggaac
6721 ctgagagaca atggcgtggg cctgaacgtg atggtgtctg agcactacca gtgctgtagg
6781 aagatcttca tccagtcttc ccacaaccag ctgaatgaga acagatggct ggagaagaca
6841 ctgaagcggg ccgagaagtg gagaagcgag ctgtccatca tgatccaggt gaagatcctg
6901 cacaccacca agtctccagc cgtgggccca aagaagaagc gcaaggtggg caccatgacc
6961 aacctgtctg acatcatcga gaaggagaca ggcaagcagc tggtaatcca ggagagcatc
7021 ctgatgctgc ctgaggaggt ggaggaggta atcggcaaca agcctgagag cgatatcctg
7081 gtgcacaccg cctatgatga gagcacagac gagaacgtga tgctgctgac cagcgacgcc
7141 cctgagtaca aaccttgggc actggtcatt caggatagta acggggagaa caaaatcaaa
7201 atgctgtgat aatgagttta aacccgctga tcagcctcga ctgtgccttc tagttgccag
7261 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
7321 gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt
7381 ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat
7441 gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctcgata
7501 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
7561 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctag
7621 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
7681 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
7741 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttctg
7801 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcatgg
7861 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
7921 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
7981 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
8041 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
8101 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
8161 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
8221 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
8281 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
8341 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
8401 ctgctgaagc cagttacctt ctgaaaaaga gttggtagct cttgatccgg caaacaaacc
8461 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
8521 tctcaagaag atcctttgat cttttctacg gggtctgaca ctcagtggaa cgaaaactca
8581 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
8641 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
8701 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
8761 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
8821 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
8881 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
8941 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
9001 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
9061 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
9121 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
9181 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
9241 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
9301 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
9361 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
9421 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
9481 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
9541 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
9601 tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg
9661 cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatcg atctcccgat
9721 cccctagggt cgactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
9781 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
9841 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
9901 cttcgcgatg tac
//
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