| 质粒名称: | v3em-Nterm-PE2max |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 207853 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 完整质粒大小: | 9279 bp |
| 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: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | PE6c/Cas9 R221K,N394K,H840A-Tf1rt P70T,G72V,S87G,M102I,K106R,K118R,I128V,L158Q,F269L,A363V,K413E,S492N,K118R,S188K,I260L,S297Q,R288Q |
| 物种来源: | Synthetic |
| 插入大小: | 5800bp |
| 基因突变: | Tf1rt(P70T, G72V, S87G, M102I, K106R, K118R, I128V, L158Q, F269L, A363V, K413E, S492N, K118R, S188K, I260L, S297Q, R288Q) |
| GenBank编号 | JN247409 |
| 启动子: | CMV |
| 标签/融合蛋白: |
SV40 bpNLS (N terminal on insert)
SV40 bpNLS, c-Myc NLS (C terminal on insert
|
| 克隆方法: | Gibson Cloning |
| 5’测序引物 | T7-forward |
| 3’测序引物 | BGH-reverse |
|
文献引用方法 |
|
| 材料和方法部分: | pCMV-PE6c was a gift from David Liu (Addgene plasmid # 207853 ; http://n2t.net/addgene:207853 ; RRID:Addgene_207853) |
| 参考文献部分: | Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Doman JL, Pandey S, Neugebauer ME, An M, Davis JR, Randolph PB, McElroy A, Gao XD, Raguram A, Richter MF, Everette KA, Banskota S, Tian K, Tao YA, Tolar J, Osborn MJ, Liu DR. Cell. 2023 Aug 31;186(18):3983-4002.e26. doi: 10.1016/j.cell.2023.07.039. 10.1016/j.cell.2023.07.039 PubMed 37657419 |
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-PE6c 9279 bp ds-DNA circular SYN 19-SEP-2023
DEFINITION Mammalian expression of PE6c prime editor.
ACCESSION .
VERSION .
KEYWORDS pCMV-PE6c
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9279)
AUTHORS Doman JL, Pandey S, Neugebauer ME, An M, Davis JR, Randolph PB,
McElroy A, Gao XD, Raguram A, Richter MF, Everette KA, Banskota S,
Tian K, Tao YA, Tolar J, Osborn MJ, Liu DR
TITLE Phage-assisted evolution and protein engineering yield compact,
efficient prime editors.
JOURNAL Cell. 2023 Aug 31;186(18):3983-4002.e26. doi:
10.1016/j.cell.2023.07.039.
PUBMED 37657419
REFERENCE 2 (bases 1 to 9279)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 19, 2023 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..9279
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(171..1031)
/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 794..813
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(1032..1136)
/gene="bla"
/label=AmpR promoter
primer_bind complement(1215..1234)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 1406..1785
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1786..1989
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1940..1960
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 2031..2050
/label=T7
/note="T7 promoter, forward primer"
promoter 2031..2049
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 2099..2119
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2120..6220
/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 6278..6298
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7883..7903
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7913..7939
/codon_start=1
/product="nuclear localization signal of human c-Myc
proto-oncogene (Dang and Lee, 1988)"
/label=c-myc NLS
/translation="PAAKRVKLD"
CDS 7948..7965
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(7988..8005)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 7994..8218
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(8289..8305)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8289..8305)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8302..8324)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8313..8329
/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(8337..8367)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8382..8403
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8520..8537)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8691..9279)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8771..8790)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
ORIGIN
1 gaagatcctt tgatcttttc tacggggtct gacactcagt ggaacgaaaa ctcacgttaa
61 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
121 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
181 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
241 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
301 atgataccgc gagaaccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
361 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
421 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
481 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
541 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
601 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
661 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
721 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
781 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
841 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
901 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
961 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
1021 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
1081 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
1141 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atcgatctcc cgatccccta
1201 gggtcgactc tcagtacaat ctgctctgat gccgcatagt taagccagta tctgctccct
1261 gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa
1321 ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc gctgcttcgc
1381 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
1441 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
1501 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
1561 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
1621 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
1681 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
1741 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
1801 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
1861 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
1921 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
1981 agcagagctg gtttagtgaa ccgtcagatc cgctagagat ccgcggccgc taatacgact
2041 cactataggg agagccgcca ccatgaaacg gacagccgac ggaagcgagt tcgagtcacc
2101 aaagaagaag cggaaagtcg acaagaagta cagcatcggc ctggacatcg gcaccaactc
2161 tgtgggctgg gccgtgatca ccgacgagta caaggtgccc agcaagaaat tcaaggtgct
2221 gggcaacacc gaccggcaca gcatcaagaa gaacctgatc ggagccctgc tgttcgacag
2281 cggcgaaaca gccgaggcca cccggctgaa gagaaccgcc agaagaagat acaccagacg
2341 gaagaaccgg atctgctatc tgcaagagat cttcagcaac gagatggcca aggtggacga
2401 cagcttcttc cacagactgg aagagtcctt cctggtggaa gaggataaga agcacgagcg
2461 gcaccccatc ttcggcaaca tcgtggacga ggtggcctac cacgagaagt accccaccat
2521 ctaccacctg agaaagaaac tggtggacag caccgacaag gccgacctgc ggctgatcta
2581 tctggccctg gcccacatga tcaagttccg gggccacttc ctgatcgagg gcgacctgaa
2641 ccccgacaac agcgacgtgg acaagctgtt catccagctg gtgcagacct acaaccagct
2701 gttcgaggaa aaccccatca acgccagcgg cgtggacgcc aaggccatcc tgtctgccag
2761 actgagcaag agcagaaagc tggaaaatct gatcgcccag ctgcccggcg agaagaagaa
2821 tggcctgttc ggaaacctga ttgccctgag cctgggcctg acccccaact tcaagagcaa
2881 cttcgacctg gccgaggatg ccaaactgca gctgagcaag gacacctacg acgacgacct
2941 ggacaacctg ctggcccaga tcggcgacca gtacgccgac ctgtttctgg ccgccaagaa
3001 cctgtccgac gccatcctgc tgagcgacat cctgagagtg aacaccgaga tcaccaaggc
3061 ccccctgagc gcctctatga tcaagagata cgacgagcac caccaggacc tgaccctgct
3121 gaaagctctc gtgcggcagc agctgcctga gaagtacaaa gagattttct tcgaccagag
3181 caagaacggc tacgccggct acattgacgg cggagccagc caggaagagt tctacaagtt
3241 catcaagccc atcctggaaa agatggacgg caccgaggaa ctgctcgtga agctgaagag
3301 agaggacctg ctgcggaagc agcggacctt cgacaacggc agcatccccc accagatcca
3361 cctgggagag ctgcacgcca ttctgcggcg gcaggaagat ttttacccat tcctgaagga
3421 caaccgggaa aagatcgaga agatcctgac cttccgcatc ccctactacg tgggccctct
3481 ggccagggga aacagcagat tcgcctggat gaccagaaag agcgaggaaa ccatcacccc
3541 ctggaacttc gaggaagtgg tggacaaggg cgcttccgcc cagagcttca tcgagcggat
3601 gaccaacttc gataagaacc tgcccaacga gaaggtgctg cccaagcaca gcctgctgta
3661 cgagtacttc accgtgtata acgagctgac caaagtgaaa tacgtgaccg agggaatgag
3721 aaagcccgcc ttcctgagcg gcgagcagaa aaaggccatc gtggacctgc tgttcaagac
3781 caaccggaaa gtgaccgtga agcagctgaa agaggactac ttcaagaaaa tcgagtgctt
3841 cgactccgtg gaaatctccg gcgtggaaga tcggttcaac gcctccctgg gcacatacca
3901 cgatctgctg aaaattatca aggacaagga cttcctggac aatgaggaaa acgaggacat
3961 tctggaagat atcgtgctga ccctgacact gtttgaggac agagagatga tcgaggaacg
4021 gctgaaaacc tatgcccacc tgttcgacga caaagtgatg aagcagctga agcggcggag
4081 atacaccggc tggggcaggc tgagccggaa gctgatcaac ggcatccggg acaagcagtc
4141 cggcaagaca atcctggatt tcctgaagtc cgacggcttc gccaacagaa acttcatgca
4201 gctgatccac gacgacagcc tgacctttaa agaggacatc cagaaagccc aggtgtccgg
4261 ccagggcgat agcctgcacg agcacattgc caatctggcc ggcagccccg ccattaagaa
4321 gggcatcctg cagacagtga aggtggtgga cgagctcgtg aaagtgatgg gccggcacaa
4381 gcccgagaac atcgtgatcg aaatggccag agagaaccag accacccaga agggacagaa
4441 gaacagccgc gagagaatga agcggatcga agagggcatc aaagagctgg gcagccagat
4501 cctgaaagaa caccccgtgg aaaacaccca gctgcagaac gagaagctgt acctgtacta
4561 cctgcagaat gggcgggata tgtacgtgga ccaggaactg gacatcaacc ggctgtccga
4621 ctacgatgtg gacgctatcg tgcctcagag ctttctgaag gacgactcca tcgacaacaa
4681 ggtgctgacc agaagcgaca agaaccgggg caagagcgac aacgtgccct ccgaagaggt
4741 cgtgaagaag atgaagaact actggcggca gctgctgaac gccaagctga ttacccagag
4801 aaagttcgac aatctgacca aggccgagag aggcggcctg agcgaactgg ataaggccgg
4861 cttcatcaag agacagctgg tggaaacccg gcagatcaca aagcacgtgg cacagatcct
4921 ggactcccgg atgaacacta agtacgacga gaatgacaag ctgatccggg aagtgaaagt
4981 gatcaccctg aagtccaagc tggtgtccga tttccggaag gatttccagt tttacaaagt
5041 gcgcgagatc aacaactacc accacgccca cgacgcctac ctgaacgccg tcgtgggaac
5101 cgccctgatc aaaaagtacc ctaagctgga aagcgagttc gtgtacggcg actacaaggt
5161 gtacgacgtg cggaagatga tcgccaagag cgagcaggaa atcggcaagg ctaccgccaa
5221 gtacttcttc tacagcaaca tcatgaactt tttcaagacc gagattaccc tggccaacgg
5281 cgagatccgg aagcggcctc tgatcgagac aaacggcgaa accggggaga tcgtgtggga
5341 taagggccgg gattttgcca ccgtgcggaa agtgctgagc atgccccaag tgaatatcgt
5401 gaaaaagacc gaggtgcaga caggcggctt cagcaaagag tctatcctgc ccaagaggaa
5461 cagcgataag ctgatcgcca gaaagaagga ctgggaccct aagaagtacg gcggcttcga
5521 cagccccacc gtggcctatt ctgtgctggt ggtggccaaa gtggaaaagg gcaagtccaa
5581 gaaactgaag agtgtgaaag agctgctggg gatcaccatc atggaaagaa gcagcttcga
5641 gaagaatccc atcgactttc tggaagccaa gggctacaaa gaagtgaaaa aggacctgat
5701 catcaagctg cctaagtact ccctgttcga gctggaaaac ggccggaaga gaatgctggc
5761 ctctgccggc gaactgcaga agggaaacga actggccctg ccctccaaat atgtgaactt
5821 cctgtacctg gccagccact atgagaagct gaagggctcc cccgaggata atgagcagaa
5881 acagctgttt gtggaacagc acaagcacta cctggacgag atcatcgagc agatcagcga
5941 gttctccaag agagtgatcc tggccgacgc taatctggac aaagtgctgt ccgcctacaa
6001 caagcaccgg gataagccca tcagagagca ggccgagaat atcatccacc tgtttaccct
6061 gaccaatctg ggagcccctg ccgccttcaa gtactttgac accaccatcg accggaagag
6121 gtacaccagc accaaagagg tgctggacgc caccctgatc caccagagca tcaccggcct
6181 gtacgagaca cggatcgacc tgtctcagct gggaggtgac tccggcggaa gctctggtgg
6241 cagcaagcgg accgccgacg gctctgaatt cgagagccct aagaagaaaa gaaaggtgag
6301 cggaggctct agcggcggaa gcatcagcag ctctaagcac accctgagcc agatgaacaa
6361 ggtgagcaac atcgtgaagg aacccgagct gcccgacatc tacaaggaat ttaaggacat
6421 caccgccgac accaataccg agaagctgcc taagcctatt aagggcctgg aatttgaagt
6481 ggaactgaca caggagaact acagactgcc tatccggaac tatcctctga ctccagtcaa
6541 gatgcaggcc atgaacgacg agatcaatca aggcctgaaa ggcggcatca tcagagagag
6601 caaggccatc aacgcctgcc ctgttatatt cgtgcccaga aaggaaggca cactgcgcat
6661 ggtggtcgac tacaggcccc tgaacaagta cgtgaagccc aacgtctacc ccctgccact
6721 gattgaacaa ctgctggcca agatccaggg cagcaccatt ttcaccaagc tggacctgaa
6781 aagcgcctac caccagatca gagtgcgaaa gggagatgag cacaagctgg ccttccggtg
6841 ccccagagga gtcttcgagt acctggtgat gccttacggc atcaagacag cccctgccca
6901 ctttcagtac ttcatcaaca caatcctggg cgaggccaag gaaagccacg tggtgtgcta
6961 catggacgac atcctgatcc attccaagtc cgagtccgaa cacgtgaaac atgtgaagga
7021 cgtgctgcaa aagctgaaga acgctaatct gatcatcaac caggccaaat gcgagtttca
7081 ccagagccaa gtgaagttcc tgggctacca catcagcgag aagggcttaa ccccatgtca
7141 ggagaacatc gacaaggtgc tgcagtggaa acagcctaaa aaccagaagg aactgagaca
7201 gttcctgggc caggtgaact acctgagaaa attcatcccc aaaaccagcc agttgaccca
7261 ccctctgaac aaactcctga aaaaggatgt cagatggaaa tggaccccta cccagaccca
7321 ggctatcgag aatatcaagc aatgtctggt gtctcctcct gtgctgaggc acttcgactt
7381 cagcaagaag atcctgcttg agacagacgt ttctgatgtg gccgtgggag ccgtgctgag
7441 ccagaagcat gatgatgata agtactaccc tgtgggctat tacagcgcta aaatgagcaa
7501 agcccagctg aattattctg tgtccgacaa ggagatgctg gctatcatca aaagcctgga
7561 gcactggcgg cactacctgg aatctacaat cgagcccttc aagatcctga ccgaccacag
7621 aaacctgatt ggaagaatca caaacgagag cgaaccagag aacaagcggc tggccagatg
7681 gcagctgttc ctgcaggact tcaacttcga gatcaactac agacctggct ctgcaaatca
7741 catcgccgat gccctgtcta gaatcgtgga cgagactgag cctatcccta aggacaacga
7801 agataacagc atcaacttcg tgaaccagat cagcatctct ggcggctcaa aaagaaccgc
7861 cgacggcagc gaattcgagt ctcccaagaa gaagaggaaa gtcggctctg gccctgccgc
7921 taagagagtg aagctggact aaccggtcat catcaccatc accattgagt ttaaacccgc
7981 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
8041 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgagaaaatt
8101 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
8161 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
8221 tctgaggcgg aaagaaccag ctggggctcg ataccgtcga cctctagcta gagcttggcg
8281 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
8341 atacgagccg gaagcataaa gtgtaaagcc tagggtgcct aatgagtgag ctaactcaca
8401 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
8461 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
8521 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
8581 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
8641 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
8701 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
8761 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
8821 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
8881 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
8941 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
9001 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
9061 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
9121 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
9181 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
9241 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaa
//
