| 质粒名称: | pET42b-BE3 |
|---|---|
| 出品公司： | addgene |
| 目录编号: | 87437 |
| 存储实验室: | David Liu |
|
载体骨架基本信息 |
|
| 载体名称: | pET_42b |
| 空载体大小: | 4083 bp |
| 完整质粒大小: | 10375bp |
| 载体类型: | 细菌细胞表达 |
| 载体抗性: | 卡那霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | BE3/GGS-His6-rAPOBEC1-XTEN-nCas9-GGS-UGI-GGS-NLS |
| 物种来源: | H. sapiens (human), R. norvegicus (rat); Bacillus phage PBS2 |
| 启动子: | T7 |
| 标签/融合蛋白: | His6 (N terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’测序引物 | TAATACGACTCACTATAGGG |
| 3’测序引物 | GCTAGTTATTGCTCAGCGG |
|
文献引用方法 |
|
| 材料和方法部分: | pET42b-BE3 was a gift from David Liu (Addgene plasmid # 87437 ; http://n2t.net/addgene:87437 ; RRID:Addgene_87437) |
| 参考文献部分: | Improving the DNA specificity and applicability of base editing through protein engineering and protein delivery. Rees HA, Komor AC, Yeh WH, Caetano-Lopes J, Warman M, Edge ASB, Liu DR. Nat Commun. 2017 Jun 6;8:15790. doi: 10.1038/ncomms15790. 10.1038/ncomms15790 PubMed 28585549 |
买家导航
pET42b-BE3载体质粒基本信息
pET42b-BE3质粒图谱载体图谱和pET42b-BE3载体序列质粒序列多克隆位点信息
pET42b-BE3质粒载体简介
pET42b-BE3质粒序列载体序列
LOCUS pET42b-BE3 10375 bp ds-DNA circular SYN 12-MAY-2021
DEFINITION Expresses BE3-NLS with an N-terminal His Tag (His6) for bacterial
expression.
ACCESSION .
VERSION .
KEYWORDS pET42b-BE3
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10375)
AUTHORS Rees HA, Komor AC, Yeh WH, Caetano-Lopes J, Warman M, Edge ASB, Liu
DR
TITLE Improving the DNA specificity and applicability of base editing
through protein engineering and protein delivery.
JOURNAL Nat Commun. 2017 Jun 6;8:15790. doi: 10.1038/ncomms15790.
PUBMED 28585549
REFERENCE 2 (bases 1 to 10375)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..10375
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS 21..38
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 39..722
/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="SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGG
RHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS
RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNE
AHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL
WATGLK"
CDS 771..4871
/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 4884..5132
/codon_start=1
/product="uracil-DNA glycosylase inhibitor from a Bacillus
subtilis bacteriophage (Mol et al., 1995)"
/label=UGI
/translation="TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTA
YDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML"
CDS 5145..5165
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
primer_bind complement(5231..5249)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 5245..5292
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
rep_origin 5329..5784
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(5416..5435)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 5626..5647
/label=F1ori-F
/note="F1 origin, forward primer"
CDS complement(5876..6691)
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin in bacteria or G418
(Geneticin(R)) in eukaryotes"
/translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind 6599..6618
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
primer_bind 6683..6702
/label=pENTR-R
/note="pENTR vectors, reverse primer"
rep_origin 6813..7401
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7302..7321
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 7555..7572
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 7587..7726
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(7652..7671)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 7812..7834
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(7828..8019)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
YELYRSCLARFGDDGENL"
CDS complement(8828..9910)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNHASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQLLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESWQ"
primer_bind 9872..9891
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(9911..9988)
/gene="lacI"
/label=lacI promoter
primer_bind 10194..10213
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 10297..10316
/label=T7
/note="T7 promoter, forward primer"
promoter 10297..10315
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 10316..10340
/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)."
RBS 10355..2
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
ORIGIN
1 gatataccat gggcagcagt catcatcatc accatcactc ttctgaaacc ggtccggttg
61 cggttgaccc gaccctgcgt cgtcgtatcg aaccgcacga attcgaagtt ttcttcgacc
121 cgcgtgaact gcgtaaagaa acctgcctgc tgtacgaaat caactggggt ggtcgtcact
181 ctatctggcg tcacacctct cagaacacca acaaacacgt tgaagttaac ttcatcgaaa
241 aattcaccac cgaacgttac ttctgcccga acacccgttg ctctatcacc tggttcctgt
301 cttggtctcc gtgcggtgaa tgctctcgtg cgatcaccga attcctgtct cgttacccgc
361 acgttaccct gttcatctac atcgcgcgtc tgtaccacca cgcggacccg cgtaaccgtc
421 agggtctgcg tgacctgatc tcttctggtg ttaccatcca gatcatgacc gaacaggaat
481 ctggttactg ctggcgtaac ttcgttaact actctccgtc taacgaagcg cactggccgc
541 gttacccgca cctgtgggtt cgtctgtacg ttctggaact gtactgcatc atcctgggtc
601 tgccgccgtg cctgaacatc ctgcgtcgta aacagccgca gctgaccttc ttcaccatcg
661 cgctgcagtc ttgccactac cagcgtctgc cgccgcacat cctgtgggcg accggtctga
721 aatccggtag cgaaacaccg gggacttcag aatcggccac cccggagtct gataagaaat
781 actcaatagg cttagctatc ggcacaaata gcgtcggatg ggcggtgatc actgatgaat
841 ataaggttcc gtctaaaaag ttcaaggttc tgggaaatac agaccgccac agtatcaaaa
901 aaaatcttat aggggctctt ttatttgaca gtggagagac agcggaagcg actcgtctca
961 aacggacagc tcgtagaagg tatacacgtc ggaagaatcg tatttgttat ctacaggaga
1021 ttttttcaaa tgagatggcg aaagtagatg atagtttctt tcatcgactt gaagagtctt
1081 ttttggtgga agaagacaag aagcatgaac gtcatcctat ttttggaaat atagtagatg
1141 aagttgctta tcatgagaaa tatccaacta tctatcatct gcgaaaaaaa ttggtagatt
1201 ctactgataa agcggatttg cgcttaatct atttggcctt agcgcatatg attaagtttc
1261 gtggtcattt tttgattgag ggagatttaa atcctgataa tagtgatgtg gacaaactat
1321 ttatccagtt ggtacaaacc tacaatcaat tatttgaaga aaaccctatt aacgcaagtg
1381 gagtagatgc taaagcgatt ctttctgcac gattgagtaa atcaagacga ttagaaaatc
1441 tcattgctca gctccccggt gagaagaaaa atggcttatt tgggaatctc attgctttgt
1501 cattgggttt gacccctaat tttaaatcaa attttgattt ggcagaagat gctaaattac
1561 agctttcaaa agatacttac gatgatgatt tagataattt attggcgcaa attggagatc
1621 aatatgctga tttgtttttg gcagctaaga atttatcaga tgctatttta ctttcagata
1681 tcctaagagt aaatactgaa ataactaagg ctcccctatc agcttcaatg attaaacgct
1741 acgatgaaca tcatcaagac ttgactcttt taaaagcttt agttcgacaa caacttccag
1801 aaaagtataa agaaatcttt tttgatcaat caaaaaacgg atatgcaggt tatattgatg
1861 ggggagctag ccaagaagaa ttttataaat ttatcaaacc aattttagaa aaaatggatg
1921 gtactgagga attattggtg aaactaaatc gtgaagattt gctgcgcaag caacggacct
1981 ttgacaacgg ctctattccc catcaaattc acttgggtga gctgcatgct attttgagaa
2041 gacaagaaga cttttatcca tttttaaaag acaatcgtga gaagattgaa aaaatcttga
2101 cttttcgaat tccttattat gttggtccat tggcgcgtgg caatagtcgt tttgcatgga
2161 tgactcggaa gtctgaagaa acaattaccc catggaattt tgaagaagtt gtcgataaag
2221 gtgcttcagc tcaatcattt attgaacgca tgacaaactt tgataaaaat cttccaaatg
2281 aaaaagtact accaaaacat agtttgcttt atgagtattt tacggtttat aacgaattga
2341 caaaggtcaa atatgttact gaaggaatgc gaaaaccagc atttctttca ggtgaacaga
2401 agaaagccat tgttgattta ctcttcaaaa caaatcgaaa agtaaccgtt aagcaattaa
2461 aagaagatta tttcaaaaaa atagaatgtt ttgatagtgt tgaaatttca ggagttgaag
2521 atagatttaa tgcttcatta ggtacctacc atgatttgct aaaaattatt aaagataaag
2581 attttttgga taatgaagaa aatgaagata tcttagagga tattgtttta acattgacct
2641 tatttgaaga tagggagatg attgaggaaa gacttaaaac atatgctcac ctctttgatg
2701 ataaggtgat gaaacagctt aaacgtcgcc gttatactgg ttggggacgt ttgtctcgaa
2761 aattgattaa tggtattagg gataagcaat ctggcaaaac aatattagat tttttgaaat
2821 cagatggttt tgccaatcgc aattttatgc agctgatcca tgatgatagt ttgacattta
2881 aagaagacat tcaaaaagca caagtgtctg gacaaggcga tagtttacat gaacatattg
2941 caaatttagc tggtagccct gctattaaaa aaggtatttt acagactgta aaagttgttg
3001 atgaattggt caaagtaatg gggcggcata agccagaaaa tatcgttatt gaaatggcac
3061 gtgaaaatca gacaactcaa aagggccaga aaaattcgcg agagcgtatg aaacgaatcg
3121 aagaaggtat caaagaatta ggaagtcaga ttcttaaaga gcatcctgtt gaaaatactc
3181 aattgcaaaa tgaaaagctc tatctctatt atctccaaaa tggaagagac atgtatgtgg
3241 accaagaatt agatattaat cgtttaagtg attatgatgt cgatcacatt gttccacaaa
3301 gtttccttaa agacgattca atagacaata aggtcttaac gcgttctgat aaaaatcgtg
3361 gtaaatcgga taacgttcca agtgaagaag tagtcaaaaa gatgaaaaac tattggagac
3421 aacttctaaa cgccaagtta atcactcaac gtaagtttga taatttaacg aaagctgaac
3481 gtggaggttt gagtgaactt gataaagctg gttttatcaa acgccaattg gttgaaactc
3541 gccaaatcac taagcatgtg gcacaaattt tggatagtcg catgaatact aaatacgatg
3601 aaaatgataa acttattcga gaggttaaag tgattacctt aaaatctaaa ttagtttctg
3661 acttccgaaa agatttccaa ttctataaag tacgtgagat taacaattac catcatgccc
3721 atgatgcgta tctaaatgcc gtcgttggaa ctgctttgat taagaaatat ccaaaacttg
3781 aatcggagtt tgtctatggt gattataaag tttatgatgt tcgtaaaatg attgctaagt
3841 ctgagcaaga aataggcaaa gcaaccgcaa aatatttctt ttactctaat atcatgaact
3901 tcttcaaaac agaaattaca cttgcaaatg gagagattcg caaacgccct ctaatcgaaa
3961 ctaatgggga aactggagaa attgtctggg ataaagggcg agattttgcc acagtgcgca
4021 aagtattgtc catgccccaa gtcaatattg tcaagaaaac agaagtacag acaggcggat
4081 tctccaagga gtcaatttta ccaaaaagaa attcggacaa gcttattgct cgtaaaaaag
4141 actgggatcc aaaaaaatat ggtggttttg atagtccaac ggtagcttat tcagtcctag
4201 ttgttgctaa ggtggaaaaa gggaaatcga agaagttaaa atccgttaaa gagttactag
4261 ggatcacaat tatggaaaga agttcctttg aaaaaaatcc gattgacttt ttagaagcta
4321 aaggatataa ggaagttaaa aaagacttaa tcattaaact acctaaatat agtctttttg
4381 agttagaaaa cggtcgtaaa cggatgctgg ctagtgccgg agaattacaa aaaggaaatg
4441 agctggctct gccaagcaaa tatgtgaatt ttttatattt agctagtcat tatgaaaagt
4501 tgaagggtag tccagaagat aacgaacaaa aacaattgtt tgtggagcag cataagcatt
4561 atttagatga gattattgag caaatcagtg aattttctaa gcgtgttatt ttagcagatg
4621 ccaatttaga taaagttctt agtgcatata acaaacatag agacaaacca atacgtgaac
4681 aagcagaaaa tattattcat ttatttacgt tgacgaatct tggagctccc gctgctttta
4741 aatattttga tacaacaatt gatcgtaaac gatatacgtc tacaaaagaa gttttagatg
4801 ccactcttat ccatcaatcc atcactggtc tttatgaaac acgcattgat ttgagtcagc
4861 taggaggtga ctctggtggt tctactaatc tgtcagatat tattgaaaag gagaccggta
4921 agcaactggt tatccaggaa tccatcctca tgctcccaga ggaggtggaa gaagtcattg
4981 ggaacaagcc ggaaagcgat atactcgtgc acaccgccta cgacgagagc accgacgaga
5041 atgtcatgct tctgactagc gacgcccctg aatacaagcc ttgggctctg gtcatacagg
5101 atagcaacgg tgagaacaag attaagatgc tctctggtgg ttctcccaag aagaagagga
5161 aagtctaata attgattaat acctaggctg ctaaacaaag cccgaaagga agctgagttg
5221 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
5281 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
5341 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
5401 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
5461 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
5521 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
5581 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
5641 aaactggaac aacactcaac catatctcgg tctattcttt tgatttataa gggattttgc
5701 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
5761 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
5821 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga
5881 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
5941 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
6001 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
6061 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
6121 ccggtgagaa tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat
6181 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
6241 gagcgagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
6301 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
6361 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
6421 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
6481 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
6541 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
6601 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag
6661 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
6721 cagacagttt tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
6781 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
6841 tgctgattgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
6901 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
6961 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
7021 ctcgctctgc taatcatgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
7081 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
7141 tcgtgcacac agccaagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
7201 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
7261 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
7321 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
7381 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
7441 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
7501 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
7561 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
7621 ggtatttcac accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa
7681 gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc
7741 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
7801 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
7861 gaggcagctg cggtaaagct cataagcgtg gtcgtgaagc gattcacaga tgtctgcctg
7921 ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa
7981 gcgggccatg ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg
8041 atttctgttc atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt
8101 tactgatgat gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg
8161 gatgcggcgg gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga
8221 tgtaggtgtt ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt
8281 gcagggcgct gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca
8341 tgttgttgct caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat
8401 cggtgattca ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga
8461 caggagcacg atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc
8521 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggagt gcaagattcc
8581 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
8641 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
8701 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
8761 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
8821 gttgcgctca ctgccagctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
8881 cggccaacgc gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca
8941 ccagtgagac gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca
9001 agcggtccac gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg
9061 ggatataaca tgagctgtct tcggtatcgt cgtatcccac taccgagata tccgcaccaa
9121 cgcgcagccc ggactcggta atggcgcgca ttgcgcccag cgccatctga tcgttggcaa
9181 ccagcatcgc agtgggaacg atgccctcat tcagcatttg catggtttgt tgaaaaccgg
9241 acatggcact ccagtcgcct tcccgttccg ctatcggctg aatttgattg cgagtgagat
9301 atttatgcca gccagccaga cgcagacgcg ccgagacaga acttaatggg cccgctaaca
9361 gcgcgatttg ctggtgaccc aatgcgacca gatgctccac gcccagtcgc gtaccgtctt
9421 catgggagaa aataatactg ttgatgggtg tctggtcaga gacatcaaga aataacgccg
9481 gaacattagt gcaggcagct tccacagcaa tggcatcctg gtcatccagc ggatagttaa
9541 tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac cgccgcttta caggcttcga
9601 cgccgcttcg ttctaccatc gacaccacca cgctggcacc cagttgatcg gcgcgagatt
9661 taatcgccgc gacaatttgc gacggcgcgt gcagggccag actggaggtg gcaacgccaa
9721 tcagcaacaa ctgtttgccc gccagttgtt gtgccacgcg gttgggaatg taattcagct
9781 ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga aacgtggctg gcatggttca
9841 ccacgcggga aacggtctga taagagacac cggcatactc tgcgacatcg tataacgtta
9901 ctggtttcac attcaccacc ctgaattgac tctcttccgg gcgctatcat gccataccgc
9961 gaaaggtttt gcgccattcg atggtgtccg ggatctcgac gctctccctt atgcgactcc
10021 tgcattagga agcagcccag tagtaggttg aggccgttga gcaccgccgc cgcaaggaat
10081 ggtgcatgca aggagatggc gcccaacagt cccccggcca cggggcctgc caccataccc
10141 acgccgaaac aagcgctcat gagcccaaag tggcgagccc gatcttcccc atcggtgatg
10201 tcggcgatat aggcgccagc aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt
10261 ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat acgactcact ataggggaat
10321 tgtgagcgga taacaattcc cctctagaaa taattttgtt taactttaag aagga
//
