| 质粒名称: | PE-Cas9 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 174851 |
| 存储实验室: | Wen Xue |
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.1 |
| 载体出品公司: | Clontech |
| 空载体大小: | 3438bp |
| 完整质粒大小: | 9753bp |
| 载体类型: | 哺乳细胞表达 |
| 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: | PE-Cas9 |
| 物种来源: | Synthetic; Cas9 is from S. pyogenes; M-MLV RT is from the Moloney murine leukemia virus |
| 启动子: | CMV |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 |
CGCAAATGGGCGGTAGGCGTG
(Common Sequencing Primers)
|
|
文献引用方法 |
|
| 材料和方法部分: | PE-Cas9 was a gift from Wen Xue (Addgene plasmid # 174851 ; http://n2t.net/addgene:174851 ; RRID:Addgene_174851) |
| 参考文献部分: | Deletion and replacement of long genomic sequences using prime editing. Jiang T, Zhang XO, Weng Z, Xue W. Nat Biotechnol. 2021 Oct 14. pii: 10.1038/s41587-021-01026-y. doi: 10.1038/s41587-021-01026-y. 10.1038/s41587-021-01026-y PubMed 34650270 |
买家导航
PE-Cas9载体质粒基本信息
PE-Cas9质粒图谱载体图谱和PE-Cas9载体序列质粒序列多克隆位点信息
PE-Cas9质粒载体简介
PE-Cas9质粒序列载体序列
LOCUS PE-Cas9 9753 bp DNA circular SYN 14-APR-2025
DEFINITION Express PE-Cas9 in mammalian cells.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9753)
AUTHORS Jiang T, Zhang XO, Weng Z, Xue W
TITLE Deletion and replacement of long genomic sequences using prime
editing.
JOURNAL Nat Biotechnol. 2021 Oct 14. pii: 10.1038/s41587-021-01026-y. doi:
10.1038/s41587-021-01026-y.
PUBMED 34650270
REFERENCE 2 (bases 1 to 9753)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2021 Oct 14. pii: 10.1038/s41587-021-01026-y. doi:
10.1038/s41587-021-01026-y."
COMMENT Sequence Label: PE-Cas9
FEATURES Location/Qualifiers
source 1..9753
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 535..555
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 694..714
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 715..4815
/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 715..4815
/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 6988..7008
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7017..7034
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7057..7074)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7063..7287
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7358..7374)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7358..7374)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7371..7393)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7382..7398
/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(join(7406..7412,7413..7430,7431..7436))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7451..7472
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7589..7606)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7760..8348)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7840..7859)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(8519..9310,9311..9379))
/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 9142..9161
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9380..9484)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9563..9582)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
601 agatccgcta gagatccgcg gccgctaata cgactcacta tagggagagc cgccaccatg
661 aaacggacag ccgacggaag cgagttcgag tcaccaaaga agaagcggaa agtcgacaag
721 aagtacagca tcggcctgga catcggcacc aactctgtgg gctgggccgt gatcaccgac
781 gagtacaagg tgcccagcaa gaaattcaag gtgctgggca acaccgaccg gcacagcatc
841 aagaagaacc tgatcggagc cctgctgttc gacagcggcg aaacagccga ggccacccgg
901 ctgaagagaa ccgccagaag aagatacacc agacggaaga accggatctg ctatctgcaa
961 gagatcttca gcaacgagat ggccaaggtg gacgacagct tcttccacag actggaagag
1021 tccttcctgg tggaagagga taagaagcac gagcggcacc ccatcttcgg caacatcgtg
1081 gacgaggtgg cctaccacga gaagtacccc accatctacc acctgagaaa gaaactggtg
1141 gacagcaccg acaaggccga cctgcggctg atctatctgg ccctggccca catgatcaag
1201 ttccggggcc acttcctgat cgagggcgac ctgaaccccg acaacagcga cgtggacaag
1261 ctgttcatcc agctggtgca gacctacaac cagctgttcg aggaaaaccc catcaacgcc
1321 agcggcgtgg acgccaaggc catcctgtct gccagactga gcaagagcag acggctggaa
1381 aatctgatcg cccagctgcc cggcgagaag aagaatggcc tgttcggaaa cctgattgcc
1441 ctgagcctgg gcctgacccc caacttcaag agcaacttcg acctggccga ggatgccaaa
1501 ctgcagctga gcaaggacac ctacgacgac gacctggaca acctgctggc ccagatcggc
1561 gaccagtacg ccgacctgtt tctggccgcc aagaacctgt ccgacgccat cctgctgagc
1621 gacatcctga gagtgaacac cgagatcacc aaggcccccc tgagcgcctc tatgatcaag
1681 agatacgacg agcaccacca ggacctgacc ctgctgaaag ctctcgtgcg gcagcagctg
1741 cctgagaagt acaaagagat tttcttcgac cagagcaaga acggctacgc cggctacatt
1801 gacggcggag ccagccagga agagttctac aagttcatca agcccatcct ggaaaagatg
1861 gacggcaccg aggaactgct cgtgaagctg aacagagagg acctgctgcg gaagcagcgg
1921 accttcgaca acggcagcat cccccaccag atccacctgg gagagctgca cgccattctg
1981 cggcggcagg aagattttta cccattcctg aaggacaacc gggaaaagat cgagaagatc
2041 ctgaccttcc gcatccccta ctacgtgggc cctctggcca ggggaaacag cagattcgcc
2101 tggatgacca gaaagagcga ggaaaccatc accccctgga acttcgagga agtggtggac
2161 aagggcgctt ccgcccagag cttcatcgag cggatgacca acttcgataa gaacctgccc
2221 aacgagaagg tgctgcccaa gcacagcctg ctgtacgagt acttcaccgt gtataacgag
2281 ctgaccaaag tgaaatacgt gaccgaggga atgagaaagc ccgccttcct gagcggcgag
2341 cagaaaaagg ccatcgtgga cctgctgttc aagaccaacc ggaaagtgac cgtgaagcag
2401 ctgaaagagg actacttcaa gaaaatcgag tgcttcgact ccgtggaaat ctccggcgtg
2461 gaagatcggt tcaacgcctc cctgggcaca taccacgatc tgctgaaaat tatcaaggac
2521 aaggacttcc tggacaatga ggaaaacgag gacattctgg aagatatcgt gctgaccctg
2581 acactgtttg aggacagaga gatgatcgag gaacggctga aaacctatgc ccacctgttc
2641 gacgacaaag tgatgaagca gctgaagcgg cggagataca ccggctgggg caggctgagc
2701 cggaagctga tcaacggcat ccgggacaag cagtccggca agacaatcct ggatttcctg
2761 aagtccgacg gcttcgccaa cagaaacttc atgcagctga tccacgacga cagcctgacc
2821 tttaaagagg acatccagaa agcccaggtg tccggccagg gcgatagcct gcacgagcac
2881 attgccaatc tggccggcag ccccgccatt aagaagggca tcctgcagac agtgaaggtg
2941 gtggacgagc tcgtgaaagt gatgggccgg cacaagcccg agaacatcgt gatcgaaatg
3001 gccagagaga accagaccac ccagaaggga cagaagaaca gccgcgagag aatgaagcgg
3061 atcgaagagg gcatcaaaga gctgggcagc cagatcctga aagaacaccc cgtggaaaac
3121 acccagctgc agaacgagaa gctgtacctg tactacctgc agaatgggcg ggatatgtac
3181 gtggaccagg aactggacat caaccggctg tccgactacg atgtggacca tatcgtgcct
3241 cagagctttc tgaaggacga ctccatcgac aacaaggtgc tgaccagaag cgacaagaac
3301 cggggcaaga gcgacaacgt gccctccgaa gaggtcgtga agaagatgaa gaactactgg
3361 cggcagctgc tgaacgccaa gctgattacc cagagaaagt tcgacaatct gaccaaggcc
3421 gagagaggcg gcctgagcga actggataag gccggcttca tcaagagaca gctggtggaa
3481 acccggcaga tcacaaagca cgtggcacag atcctggact cccggatgaa cactaagtac
3541 gacgagaatg acaagctgat ccgggaagtg aaagtgatca ccctgaagtc caagctggtg
3601 tccgatttcc ggaaggattt ccagttttac aaagtgcgcg agatcaacaa ctaccaccac
3661 gcccacgacg cctacctgaa cgccgtcgtg ggaaccgccc tgatcaaaaa gtaccctaag
3721 ctggaaagcg agttcgtgta cggcgactac aaggtgtacg acgtgcggaa gatgatcgcc
3781 aagagcgagc aggaaatcgg caaggctacc gccaagtact tcttctacag caacatcatg
3841 aactttttca agaccgagat taccctggcc aacggcgaga tccggaagcg gcctctgatc
3901 gagacaaacg gcgaaaccgg ggagatcgtg tgggataagg gccgggattt tgccaccgtg
3961 cggaaagtgc tgagcatgcc ccaagtgaat atcgtgaaaa agaccgaggt gcagacaggc
4021 ggcttcagca aagagtctat cctgcccaag aggaacagcg ataagctgat cgccagaaag
4081 aaggactggg accctaagaa gtacggcggc ttcgacagcc ccaccgtggc ctattctgtg
4141 ctggtggtgg ccaaagtgga aaagggcaag tccaagaaac tgaagagtgt gaaagagctg
4201 ctggggatca ccatcatgga aagaagcagc ttcgagaaga atcccatcga ctttctggaa
4261 gccaagggct acaaagaagt gaaaaaggac ctgatcatca agctgcctaa gtactccctg
4321 ttcgagctgg aaaacggccg gaagagaatg ctggcctctg ccggcgaact gcagaaggga
4381 aacgaactgg ccctgccctc caaatatgtg aacttcctgt acctggccag ccactatgag
4441 aagctgaagg gctcccccga ggataatgag cagaaacagc tgtttgtgga acagcacaag
4501 cactacctgg acgagatcat cgagcagatc agcgagttct ccaagagagt gatcctggcc
4561 gacgctaatc tggacaaagt gctgtccgcc tacaacaagc accgggataa gcccatcaga
4621 gagcaggccg agaatatcat ccacctgttt accctgacca atctgggagc ccctgccgcc
4681 ttcaagtact ttgacaccac catcgaccgg aagaggtaca ccagcaccaa agaggtgctg
4741 gacgccaccc tgatccacca gagcatcacc ggcctgtacg agacacggat cgacctgtct
4801 cagctgggag gtgactctgg aggatctagc ggaggatcct ctggcagcga gacaccagga
4861 acaagcgagt cagcaacacc agagagcagt ggcggcagca gcggcggcag cagcacccta
4921 aatatagaag atgagtatcg gctacatgag acctcaaaag agccagatgt ttctctaggg
4981 tccacatggc tgtctgattt tcctcaggcc tgggcggaaa ccgggggcat gggactggca
5041 gttcgccaag ctcctctgat catacctctg aaagcaacct ctacccccgt gtccataaaa
5101 caatacccca tgtcacaaga agccagactg gggatcaagc cccacataca gagactgttg
5161 gaccagggaa tactggtacc ctgccagtcc ccctggaaca cgcccctgct acccgttaag
5221 aaaccaggga ctaatgatta taggcctgtc caggatctga gagaagtcaa caagcgggtg
5281 gaagacatcc accccaccgt gcccaaccct tacaacctct tgagcgggct cccaccgtcc
5341 caccagtggt acactgtgct tgatttaaag gatgcctttt tctgcctgag actccacccc
5401 accagtcagc ctctcttcgc ctttgagtgg agagatccag agatgggaat ctcaggacaa
5461 ttgacctgga ccagactccc acagggtttc aaaaacagtc ccaccctgtt taatgaggca
5521 ctgcacagag acctagcaga cttccggatc cagcacccag acttgatcct gctacagtac
5581 gtggatgact tactgctggc cgccacttct gagctagact gccaacaagg tactcgggcc
5641 ctgttacaaa ccctagggaa cctcgggtat cgggcctcgg ccaagaaagc ccaaatttgc
5701 cagaaacagg tcaagtatct ggggtatctt ctaaaagagg gtcagagatg gctgactgag
5761 gccagaaaag agactgtgat ggggcagcct actccgaaga cccctcgaca actaagggag
5821 ttcctaggga aggcaggctt ctgtcgcctc ttcatccctg ggtttgcaga aatggcagcc
5881 cccctgtacc ctctcaccaa accggggact ctgtttaatt ggggcccaga ccaacaaaag
5941 gcctatcaag aaatcaagca agctcttcta actgccccag ccctggggtt gccagatttg
6001 actaagccct ttgaactctt tgtcgacgag aagcagggct acgccaaagg tgtcctaacg
6061 caaaaactgg gaccttggcg tcggccggtg gcctacctgt ccaaaaagct agacccagta
6121 gcagctgggt ggcccccttg cctacggatg gtagcagcca ttgccgtact gacaaaggat
6181 gcaggcaagc taaccatggg acagccacta gtcattctgg ccccccatgc agtagaggca
6241 ctagtcaaac aaccccccga ccgctggctt tccaacgccc ggatgactca ctatcaggcc
6301 ttgcttttgg acacggaccg ggtccagttc ggaccggtgg tagccctgaa cccggctacg
6361 ctgctcccac tgcctgagga agggctgcaa cacaactgcc ttgatatcct ggccgaagcc
6421 cacggaaccc gacccgacct aacggaccag ccgctcccag acgccgacca cacctggtac
6481 acggatggaa gcagtctctt acaagaggga cagcgtaagg cgggagctgc ggtgaccacc
6541 gagaccgagg taatctgggc taaagccctg ccagccggga catccgctca gcgggctgaa
6601 ctgatagcac tcacccaggc cctaaagatg gcagaaggta agaagctaaa tgtttatact
6661 gatagccgtt atgcttttgc tactgcccat atccatggag aaatatacag aaggcgtggg
6721 tggctcacat cagaaggcaa agagatcaaa aataaagacg agatcttggc cctactaaaa
6781 gccctctttc tgcccaaaag acttagcata atccattgtc caggacatca aaagggacac
6841 agcgccgagg ctagaggcaa ccggatggct gaccaagcgg cccgaaaggc agccatcaca
6901 gagactccag acacctctac cctcctcata gaaaattcat caccctctgg cggctcaaaa
6961 agaaccgccg acggcagcga attcgagccc aagaagaaga ggaaagtcta accggtcatc
7021 atcaccatca ccattgagtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
7081 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
7141 ctgtcctttc ctaataaaat gagaaaattg catcgcattg tctgagtagg tgtcattcta
7201 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
7261 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcga
7321 taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
7381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
7441 agggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
7501 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
7561 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
7621 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
7681 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
7741 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
7801 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
7861 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
7921 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
7981 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
8041 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
8101 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
8161 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
8221 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
8281 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
8341 gatctcaaga agatcctttg atcttttcta cggggtctga cactcagtgg aacgaaaact
8401 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
8461 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
8521 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
8581 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
8641 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
8701 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
8761 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
8821 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
8881 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
8941 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
9001 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
9061 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
9121 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
9181 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
9241 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
9301 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
9361 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
9421 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
9481 cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat cgatctcccg
9541 atcccctagg gtctactctc agtacaatct gctctgatgc cgcatagtta agccagtatc
9601 tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa
9661 caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg cgttttgcgc
9721 tgcttcgcga tgtacgggcc agatatacgc gtt
//
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