| 质粒名称: | pT7-PEmax for IVT |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 178113 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pT7 |
| 载体出品公司: | Clontech |
| 完整质粒大小: | 9874bp |
| 载体类型: | Template for in vitro transcription |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PEmax / SV40_bpNLS-SpCas9(R221K,N394K,H840A)-SGGSx2-SV40_bpNLS-SGGSx2-MMLV_RT(D200N, T306K, W313F, T330P, L603W)-SV40_bpNLS-cMyc_NLS |
| 物种来源: | Synthetic; SpCas9 is from Streptococcus pyogenes; MMLV_RT is from the Moloney murine leukemia virus |
| 插入大小: | 6393bp |
| 基因突变: | Detailed in manuscript |
| GenBank编号 | JN247409 |
| 启动子: | T7 (inactivated) |
| 标签/融合蛋白: |
SV40 bpNLS (N terminal on insert)
SV40 bpNLS (C terminal on insert)
c-Myc NLS (C terminal on insert)
|
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | CMV-F |
| 3’测序引物 | BGH-reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pT7-PEmax for IVT was a gift from David Liu (Addgene plasmid # 178113 ; http://n2t.net/addgene:178113 ; RRID:Addgene_178113) |
| 参考文献部分: | Enhanced prime editing systems by manipulating cellular determinants of editing outcomes. Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B, Liu DR. Cell. 2021 Oct 28;184(22):5635-5652.e29. doi: 10.1016/j.cell.2021.09.018. 10.1016/j.cell.2021.09.018 PubMed 34653350 |
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 pT7-PEmax for IVT 9874 bp ds-DNA circular SYN 30-NOV-2021
DEFINITION Template for in vitro transcription of PEmax.
ACCESSION .
VERSION .
KEYWORDS pT7-PEmax for IVT
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9874)
AUTHORS Chen PJ, Hussmann JA, Yan J, Knipping F, Ravisankar P, Chen PF, Chen
C, Nelson JW, Newby GA, Sahin M, Osborn MJ, Weissman JS, Adamson B,
Liu DR
TITLE Enhanced prime editing systems by manipulating cellular determinants
of editing outcomes.
JOURNAL Cell. 2021 Oct 9. pii: S0092-8674(21)01065-5. doi:
10.1016/j.cell.2021.09.018.
PUBMED 34653350
REFERENCE 2 (bases 1 to 9874)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 30, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9874
/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"
CDS 340..360
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 361..4461
/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
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRKLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLKREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 4519..4539
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6640..6660
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6670..6696
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
polyA_signal 6804..7028
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(7099..7115)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(7099..7115)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(7112..7134)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 7123..7139
/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(7147..7177)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 7192..7213
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(7330..7347)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(7501..8089)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7581..7600)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(8260..9120)
/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 8883..8902
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(9121..9225)
/gene="bla"
/label=AmpR promoter
primer_bind complement(9304..9323)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 9495..9874
/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 agatctcgag ctcggtacct
241 aatacgacac actataagga aataagagag aaaagaagag taagaagaaa tataagagcc
301 accatgaaac ggacagccga cggaagcgag ttcgagtcac caaagaagaa gcggaaagtc
361 gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg ggccgtgatc
421 accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac cgaccggcac
481 agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac agccgaggcc
541 acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg gatctgctat
601 ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt ccacagactg
661 gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat cttcggcaac
721 atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct gagaaagaaa
781 ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct ggcccacatg
841 atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa cagcgacgtg
901 gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga aaaccccatc
961 aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa gagcagaaag
1021 ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt cggaaacctg
1081 attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct ggccgaggat
1141 gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct gctggcccag
1201 atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga cgccatcctg
1261 ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag cgcctctatg
1321 atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct cgtgcggcag
1381 cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg ctacgccggc
1441 tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc catcctggaa
1501 aagatggacg gcaccgagga actgctcgtg aagctgaaga gagaggacct gctgcggaag
1561 cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga gctgcacgcc
1621 attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga aaagatcgag
1681 aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg aaacagcaga
1741 ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt cgaggaagtg
1801 gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt cgataagaac
1861 ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt caccgtgtat
1921 aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc cttcctgagc
1981 ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa agtgaccgtg
2041 aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt ggaaatctcc
2101 ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct gaaaattatc
2161 aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga tatcgtgctg
2221 accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac ctatgcccac
2281 ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg ctggggcagg
2341 ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac aatcctggat
2401 ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca cgacgacagc
2461 ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga tagcctgcac
2521 gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct gcagacagtg
2581 aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa catcgtgatc
2641 gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg cgagagaatg
2701 aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga acaccccgtg
2761 gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa tgggcgggat
2821 atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt ggacgctatc
2881 gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac cagaagcgac
2941 aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa gatgaagaac
3001 tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga caatctgacc
3061 aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa gagacagctg
3121 gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg gatgaacact
3181 aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct gaagtccaag
3241 ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat caacaactac
3301 caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat caaaaagtac
3361 cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt gcggaagatg
3421 atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt ctacagcaac
3481 atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg gaagcggcct
3541 ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg ggattttgcc
3601 accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac cgaggtgcag
3661 acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa gctgatcgcc
3721 agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac cgtggcctat
3781 tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa gagtgtgaaa
3841 gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc catcgacttt
3901 ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct gcctaagtac
3961 tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg cgaactgcag
4021 aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct ggccagccac
4081 tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt tgtggaacag
4141 cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa gagagtgatc
4201 ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg ggataagccc
4261 atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct gggagcccct
4321 gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag caccaaagag
4381 gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac acggatcgac
4441 ctgtctcagc tgggaggtga ctccggcgga agctctggtg gcagcaagcg gaccgccgac
4501 ggctctgaat tcgagagccc taagaagaaa agaaaggtga gcggaggctc tagcggcgga
4561 agcaccctga acattgaaga cgagtataga ctgcatgaaa caagcaagga acccgatgtg
4621 tccctgggct ccacctggct gtccgacttt ccccaggcct gggccgagac aggaggaatg
4681 ggcctggccg tgcggcaggc acccctgatc atccctctga aggccacctc tacacccgtg
4741 agcatcaagc agtaccctat gtctcaggag gccagactgg gcatcaagcc tcacatccag
4801 aggctgctgg accagggcat cctggtgcca tgccagagcc cctggaacac accactgctg
4861 cccgtgaaga agccaggcac caatgactat agacccgtgc aggatctgag agaggtgaac
4921 aagagggtgg aggatatcca ccccaccgtg cccaaccctt acaatctgct gtccggcctg
4981 cccccttctc accagtggta tacagtgctg gacctgaagg atgccttctt ttgtctgaga
5041 ctgcacccta ccagccagcc actgttcgcc tttgagtgga gggaccctga gatgggcatc
5101 tctggccagc tgacctggac acgcctgcct cagggcttca agaatagccc aacactgttt
5161 aacgaggccc tgcaccgcga cctggcagat ttccggatcc agcacccaga tctgatcctg
5221 ctgcagtacg tggacgatct gctgctggcc gccaccagcg agctggattg ccagcaggga
5281 acacgcgccc tgctgcagac cctgggaaac ctgggatata gggcatccgc caagaaggcc
5341 cagatctgtc agaagcaggt gaagtacctg ggctatctgc tgaaggaggg ccagagatgg
5401 ctgacagagg ccaggaagga gacagtgatg ggccagccaa cacccaagac cccaagacag
5461 ctgagggagt tcctgggcaa agcaggattt tgcaggctgt tcatcccagg attcgcagag
5521 atggcagcac ctctgtaccc actgaccaag ccgggcaccc tgtttaattg gggccctgac
5581 cagcagaagg cctatcagga gatcaagcag gccctgctga cagcaccagc cctgggcctg
5641 ccagacctga ccaagccttt cgagctgttt gtggatgaga agcagggcta cgccaagggc
5701 gtgctgaccc agaagctggg accatggaga cggcccgtgg cctatctgtc caagaagctg
5761 gacccagtgg cagcaggatg gccaccatgc ctgaggatgg tggcagcaat cgccgtgctg
5821 acaaaggatg ccggcaagct gaccatggga cagccactgg tcatcctggc accacacgca
5881 gtggaggccc tggtgaagca gcctccagat cgctggctgt ctaacgcccg gatgacacac
5941 taccaggccc tgctgctgga caccgatcgc gtgcagtttg gccctgtggt ggccctgaat
6001 ccagccaccc tgctgcctct gccagaggag ggcctgcagc acaactgtct ggacatcctg
6061 gcagaggcac acggaacaag gccagacctg accgatcagc ccctgcctga cgccgatcac
6121 acatggtata ccgatggaag ctccctgctg caggagggcc agaggaaggc aggagcagca
6181 gtgaccacag agacagaagt gatctgggcc aaggccctgc cagcaggcac atccgcccag
6241 cgggccgagc tgatcgccct gacccaggcc ctgaagatgg ccgagggcaa gaagctgaac
6301 gtgtacacag actccagata tgccttcgcc accgcacaca tccacggaga gatctacagg
6361 cgccggggct ggctgacctc tgagggcaag gagatcaaga acaaggatga gatcctggcc
6421 ctgctgaagg ccctgtttct gcccaagcgg ctgagcatca tccactgtcc tggacaccag
6481 aagggacact ccgccgaggc aaggggcaat cggatggccg accaggccgc cagaaaggct
6541 gctattactg aaactcccga cacttccact ctgctgattg aaaactcctc cccttctggc
6601 ggctcaaaaa gaaccgccga cggcagcgaa ttcgagtctc ccaagaagaa gaggaaagtc
6661 ggctctggcc ctgccgctaa gagagtgaag ctggactaat taattaagct gccttctgcg
6721 gggcttgcct tctggccatg cccttcttct ctcccttgca cctgtacctc ttggtctttg
6781 aataaagcct gagtaggaag cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
6841 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
6901 tgagaaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
6961 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
7021 ctctatggct tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta
7081 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
7141 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag
7201 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
7261 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
7321 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
7381 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
7441 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
7501 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
7561 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
7621 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
7681 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
7741 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
7801 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
7861 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
7921 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
7981 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
8041 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
8101 gatcttttct acggggtctg acactcagtg gaacgaaaac tcacgttaag ggattttggt
8161 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
8221 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
8281 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
8341 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
8401 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
8461 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
8521 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
8581 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
8641 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
8701 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
8761 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
8821 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
8881 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
8941 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
9001 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
9061 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
9121 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
9181 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
9241 agtgccacct gacgtcgacg gatcgggaga tcgatctccc gatcccctag ggtctactct
9301 cagtacaatc tgctctgatg ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt
9361 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
9421 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
9481 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
9541 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
9601 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
9661 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
9721 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
9781 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
9841 gtacatctac gtattagtca tcgctattac catg
//
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