| 质粒名称: | xCas9(3.7)-BE4 |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 108381 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pCMV |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | xCas9(3.7)-BE4,rAPOBEC1-XTEN-nxCas9(3.7)-UGI-UGI-NLS |
| 物种来源: | Synthetic |
| 插入大小: | 5478 bp |
| 基因突变: | A262T, R324L, S409I, E480K, E543D, M694I, E1219V |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | NLS（插入件上的N端子） |
| 克隆方法: | Gibson克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV Forward |
| 3' 测序引物 | M13 |
|
文献引用方法 |
|
| 材料和方法部分: | xCas9(3.7)-BE4 was a gift from David Liu (Addgene plasmid # 108381 ; http://n2t.net/addgene:108381 ; RRID:Addgene_108381) |
| 参考文献部分: | Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X, Rees HA, Lin Z, Liu DR. Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155. 10.1038/nature26155 PubMed 29512652 |
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
-BE4质粒图谱.png)
载体简介
载体序列
LOCUS xCas9(3.7)-BE4 8877 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Mammalian expression vector for xCas9(3.7)-BE4.
ACCESSION .
VERSION .
KEYWORDS xCas9(3.7)-BE4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8877)
AUTHORS Hu JH, Miller SM, Geurts MH, Tang W, Chen L, Sun N, Zeina CM, Gao X,
Rees HA, Lin Z, Liu DR
TITLE Evolved Cas9 variants with broad PAM compatibility and high DNA
specificity.
JOURNAL Nature. 2018 Feb 28. pii: nature26155. doi: 10.1038/nature26155.
PUBMED 29512652
REFERENCE 2 (bases 1 to 8877)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8877
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 74..453
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 454..657
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 608..628
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
primer_bind 699..718
/label=T7
/note="T7 promoter, forward primer"
promoter 699..717
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 731..1417
/codon_start=1
/product="cytidine deaminase (C to U editing enzyme) from
rat"
/label=APOBEC-1
/note="can use ssDNA as a substrate (Komor et al., 2016)"
/translation="MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWG
GRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL
SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSN
EAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI
LWATGLK"
CDS 1514..5614
/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="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDTKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKLYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGIIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEKVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGDQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFIQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGVLQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 5645..5893
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5924..6172
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 6185..6205
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6214..6231
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
primer_bind complement(6254..6271)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 6260..6484
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(6555..6571)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6555..6571)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(6568..6590)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 6579..6595
/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(6603..6633)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 6648..6669
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(6786..6803)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(6957..7545)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7037..7056)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(7716..8576)
/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 8339..8358
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(8577..8681)
/gene="bla"
/label=AmpR promoter
primer_bind complement(8760..8779)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
ORIGIN
1 aattgcatga agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc
61 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
121 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
181 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
241 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
301 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
361 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
421 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
481 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
541 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
601 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctggt
661 ttagtgaacc gtcagatccg ctagagatcc gcggccgcta atacgactca ctatagggag
721 agccgccacc atgagctcag agactggccc agtggctgtg gaccccacat tgagacggcg
781 gatcgagccc catgagtttg aggtattctt cgatccgaga gagctccgca aggagacctg
841 cctgctttac gaaattaatt gggggggccg gcactccatt tggcgacata catcacagaa
901 cactaacaag cacgtcgaag tcaacttcat cgagaagttc acgacagaaa gatatttctg
961 tccgaacaca aggtgcagca ttacctggtt tctcagctgg agcccatgcg gcgaatgtag
1021 tagggccatc actgaattcc tgtcaaggta tccccacgtc actctgttta tttacatcgc
1081 aaggctgtac caccacgctg acccccgcaa tcgacaaggc ctgcgggatt tgatctcttc
1141 aggtgtgact atccaaatta tgactgagca ggagtcagga tactgctgga gaaactttgt
1201 gaattatagc ccgagtaatg aagcccactg gcctaggtat ccccatctgt gggtacgact
1261 gtacgttctt gaactgtact gcatcatact gggcctgcct ccttgtctca acattctgag
1321 aaggaagcag ccacagctga cattctttac catcgctctt cagtcttgtc attaccagcg
1381 actgccccca cacattctct gggccaccgg gttgaaatct ggtggttctt ctggtggttc
1441 tagcggcagc gagactcccg ggacctcaga gtccgccaca cccgaaagtt ctggtggttc
1501 ttctggtggt tctgacaaga agtactccat tgggctcgct atcggcacaa acagcgtcgg
1561 ctgggccgtc attacggacg agtacaaggt gccgagcaaa aaattcaaag ttctgggcaa
1621 taccgatcgc cacagcataa agaagaacct cattggcgcc ctcctgttcg actccgggga
1681 gacggccgaa gccacgcggc tcaaaagaac agcacggcgc agatataccc gcagaaagaa
1741 tcggatctgc tacctgcagg agatctttag taatgagatg gctaaggtgg atgactcttt
1801 cttccatagg ctggaggagt cctttttggt ggaggaggat aaaaagcacg agcgccaccc
1861 aatctttggc aatatcgtgg acgaggtggc gtaccatgaa aagtacccaa ccatatatca
1921 tctgaggaag aagcttgtag acagtactga taaggctgac ttgcggttga tctatctcgc
1981 gctggcgcat atgatcaaat ttcggggaca cttcctcatc gagggggacc tgaacccaga
2041 caacagcgat gtcgacaaac tctttatcca actggttcag acttacaatc agcttttcga
2101 agagaacccg atcaacgcat ccggagttga cgccaaagca atcctgagcg ctaggctgtc
2161 caaatcccgg cggctcgaaa acctcatcgc acagctccct ggggagaaga agaacggcct
2221 gtttggtaat cttatcgccc tgtccctcgg gctgaccccc aactttaaat ctaacttcga
2281 cctggccgaa gataccaagc ttcaactgag caaagacacc tacgatgatg atctcgacaa
2341 tctgctggcc cagatcggcg accagtacgc agaccttttt ttggcggcaa agaacctgtc
2401 agacgccatt ctgctgagtg atattctgcg agtgaacacg gagatcacca aagctccgct
2461 gagcgctagt atgatcaagc tctatgatga gcaccaccaa gacttgactt tgctgaaggc
2521 ccttgtcaga cagcaactgc ctgagaagta caaggaaatt ttcttcgatc agtctaaaaa
2581 tggctacgcc ggatacattg acggcggagc aagccaggag gaattttaca aatttattaa
2641 gcccatcttg gaaaaaatgg acggcaccga ggagctgctg gtaaagctta acagagaaga
2701 tctgttgcgc aaacagcgca ctttcgacaa tggaatcatc ccccaccaga ttcacctggg
2761 cgaactgcac gctatcctca ggcggcaaga ggatttctac ccctttttga aagataacag
2821 ggaaaagatt gagaaaatcc tcacatttcg gataccctac tatgtaggcc ccctcgcccg
2881 gggaaattcc agattcgcgt ggatgactcg caaatcagaa gagaccatca ctccctggaa
2941 cttcgagaaa gtcgtggata agggggcctc tgcccagtcc ttcatcgaaa ggatgactaa
3001 ctttgataaa aatctgccta acgaaaaggt gcttcctaaa cactctctgc tgtacgagta
3061 cttcacagtt tataacgagc tcaccaaggt caaatacgtc acagaaggga tgagaaagcc
3121 agcattcctg tctggagatc agaagaaagc tattgtggac ctcctcttca agacgaaccg
3181 gaaagttacc gtgaaacagc tcaaagaaga ctatttcaaa aagattgaat gtttcgactc
3241 tgttgaaatc agcggagtgg aggatcgctt caacgcatcc ctgggaacgt atcacgatct
3301 cctgaaaatc attaaagaca aggacttcct ggacaatgag gagaacgagg acattcttga
3361 ggacattgtc ctcaccctta cgttgtttga agatagggag atgattgaag aacgcttgaa
3421 aacttacgct catctcttcg acgacaaagt catgaagcag ctcaagaggc gccgatatac
3481 aggatggggg cggctgtcaa gaaaactgat caatgggatc cgagacaagc agagtggaaa
3541 gacaatcctg gattttctta agtccgatgg atttgccaac cggaacttca ttcagttgat
3601 ccatgatgac tctctcacct ttaaggagga catccagaaa gcacaagttt ctggccaggg
3661 ggacagtctt cacgagcaca tcgctaatct tgcaggtagc ccagctatca aaaagggaat
3721 actgcagacc gttaaggtcg tggatgaact cgtcaaagta atgggaaggc ataagcccga
3781 gaatatcgtt atcgagatgg cccgagagaa ccaaaccacc cagaagggac agaagaacag
3841 tagggaaagg atgaagagga ttgaagaggg tataaaagaa ctggggtccc aaatccttaa
3901 ggaacaccca gttgaaaaca cccagcttca gaatgagaag ctctacctgt actacctgca
3961 gaacggcagg gacatgtacg tggatcagga actggacatc aatcggctct ccgactacga
4021 cgtggatcat atcgtgcccc agtcttttct caaagatgat tctattgata ataaagtgtt
4081 gacaagatcc gataaaaaca gagggaagag tgataacgtc ccctcagaag aagttgtcaa
4141 gaaaatgaaa aattattggc ggcagctgct gaacgccaaa ctgatcacac aacggaagtt
4201 cgataatctg actaaggctg aacgaggtgg cctgtctgag ttggataaag ccggtttcat
4261 caaaaggcag cttgttgaga cacgccagat caccaagcac gtggcccaaa ttctcgattc
4321 acgcatgaac accaagtacg atgaaaatga caaactgatt cgagaggtga aagttattac
4381 tctgaagtct aagctggtct cagatttcag aaaggacttt cagttttata aggtgagaga
4441 gatcaacaat taccaccatg cgcatgatgc ctacctgaat gcagtggtag gcactgcact
4501 tatcaaaaaa tatcccaagc ttgaatctga atttgtttac ggagactata aagtgtacga
4561 tgttaggaaa atgatcgcaa agtctgagca ggaaataggc aaggccaccg ctaagtactt
4621 cttttacagc aatattatga attttttcaa gaccgagatt acactggcca atggagagat
4681 tcggaagcga ccacttatcg aaacaaacgg agaaacagga gaaatcgtgt gggacaaggg
4741 tagggatttc gcgacagtcc ggaaggtcct gtccatgccg caggtgaaca tcgttaaaaa
4801 gaccgaagta cagaccggag gcttctccaa ggaaagtatc ctcccgaaaa ggaacagcga
4861 caagctgatc gcacgcaaaa aagattggga ccccaagaaa tacggcggat tcgattctcc
4921 tacagtcgct tacagtgtac tggttgtggc taaagtggag aaagggaagt ctaaaaaact
4981 caaaagcgtc aaggaactgc tgggcatcac aatcatggag cgatcaagct tcgaaaaaaa
5041 ccccatcgac tttctcgagg cgaaaggata taaagaggtc aaaaaagacc tcatcattaa
5101 gcttcccaag tactctctct ttgagcttga aaacggccgg aaacgaatgc tcgctagtgc
5161 gggcgtgctg cagaaaggta acgagctggc actgccctct aaatacgtta atttcttgta
5221 tctggccagc cactatgaaa agctcaaagg gtctcccgaa gataatgagc agaagcagct
5281 gttcgtggaa caacacaaac actaccttga tgagatcatc gagcaaataa gcgaattctc
5341 caaaagagtg atcctcgccg acgctaacct cgataaggtg ctttctgctt acaataagca
5401 cagggataag cccatcaggg agcaggcaga aaacattatc cacttgttta ctctgaccaa
5461 cttgggcgcg cctgcagcct tcaagtactt cgacactacc atagacagaa agcggtacac
5521 ctctacaaag gaggtcctgg acgccacact gattcatcag tcaattacgg ggctctatga
5581 aacaagaatc gacctctctc agctcggtgg agactctggt ggttctggag gatctggtgg
5641 ttctactaat ctgtcagata ttattgaaaa ggagaccggt aagcaactgg ttatccagga
5701 atccatcctc atgctcccag aggaggtgga agaagtcatt gggaacaagc cggaaagcga
5761 tatactcgtg cacaccgcct acgacgagag caccgacgag aatgtcatgc ttctgactag
5821 cgacgcccct gaatacaagc cttgggctct ggtcatacag gatagcaacg gtgagaacaa
5881 gattaagatg ctctctggtg gttctggagg atctggtggt tctactaatc tgtcagatat
5941 tattgaaaag gagaccggta agcaactggt tatccaggaa tccatcctca tgctcccaga
6001 ggaggtggaa gaagtcattg ggaacaagcc ggaaagcgat atactcgtgc acaccgccta
6061 cgacgagagc accgacgaga atgtcatgct tctgactagc gacgcccctg aatacaagcc
6121 ttgggctctg gtcatacagg atagcaacgg tgagaacaag attaagatgc tctctggtgg
6181 ttctcccaag aagaagagga aagtctaacc ggtcatcatc accatcacca ttgagtttaa
6241 acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt ttgcccctcc
6301 cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta ataaaatgag
6361 gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg ggtggggcag
6421 gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc ggtgggctct
6481 atggcttctg aggcggaaag aaccagctgg ggctcgatac cgtcgacctc tagctagagc
6541 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
6601 cacaacatac gagccggaag cataaagtgt aaagcctagg gtgcctaatg agtgagctaa
6661 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
6721 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
6781 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
6841 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
6901 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
6961 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
7021 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
7081 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
7141 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
7201 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
7261 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
7321 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
7381 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
7441 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
7501 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
7561 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
7621 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
7681 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
7741 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
7801 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
7861 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
7921 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
7981 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
8041 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
8101 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
8161 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
8221 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
8281 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
8341 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
8401 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
8461 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
8521 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
8581 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
8641 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
8701 ccacctgacg tcgacggatc gggagatcga tctcccgatc ccctagggtc gactctcagt
8761 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
8821 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgac
//
