| 质粒名称: | SP-dCas9-VPR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 63798 |
| 存储实验室: |
George Church
|
|
载体骨架基本信息 |
|
| 载体名称: | pcDNA3.3 TOPO |
| 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: | 哺乳动物表达，CRISPR，合成生物学 |
| 筛选标记: | 新霉素（G418） |
| 高拷贝/低拷贝: | -- |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | SP-dCas9-VPR,VP64-p65-Rta |
| 物种来源: | H. sapiens (human), M. musculus (mouse) |
| 插入大小: | 5799 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | CMV |
| 标签/融合蛋白: | VPR（VP64-p65-Rta）（插入的C端子） |
| 克隆方法: | Gateway 克隆 |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | SP-dCas9-VPR was a gift from George Church (Addgene plasmid # 63798 ; http://n2t.net/addgene:63798 ; RRID:Addgene_63798) |
| 参考文献部分: | Highly efficient Cas9-mediated transcriptional programming. Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan D, Braff JL, Davidsohn N, Housden BE, Perrimon N, Weiss R, Aach J, Collins JJ, Church GM. Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312. 10.1038/nmeth.3312 PubMed 25730490 |
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 SP-dCas9-VPR 11319 bp ds-DNA circular SYN 30-SEP-2019
DEFINITION SP-dCas9 with VP64-p65-Rta (VPR) fused to it's C-terminus; mammalian
vector.
ACCESSION .
VERSION .
KEYWORDS SP-dCas9-VPR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11319)
AUTHORS Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin
S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan D, Braff JL,
Davidsohn N, Housden BE, Perrimon N, Weiss R, Aach J, Collins JJ,
Church GM
TITLE Highly efficient Cas9-mediated transcriptional programming.
JOURNAL Nat Methods. 2015 Mar 2. doi: 10.1038/nmeth.3312.
PUBMED 25730490
REFERENCE 2 (bases 1 to 11319)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Sep 30, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11319
/organism="synthetic DNA construct"
/mol_type="other DNA"
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 581..605
/label=LNCX
/note="Human CMV promoter, forward primer"
regulatory 693..702
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 699..4802
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=Cas9m4
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the mutations D10A, D839A,
H840A, and N863A (Mali et al., 2013)"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 4815..4835
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
protein_bind 4872..4896
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS 4926..5075
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
CDS 5100..5120
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5550..5906
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALL"
CDS 5925..6494
/codon_start=1
/product="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
/label=Rta AD
/translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
DTFLNDECLLHAMHISTGLSIFDTSLF"
protein_bind complement(6499..6523)
/gene="mutant version of attB"
/label=attB2
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
promoter complement(6573..6590)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(6656..6675)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 6700..6748
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 6950..7378
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(7037..7056)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 7247..7268
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(7387..7407)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 7392..7721
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 7572..7707
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 7634..7653
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 7788..8582
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7842..7861)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8452..8471
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 8758..8879
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8795..8814)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8849..8868
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(8928..8944)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8928..8944)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8941..8963)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8952..8968
/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(8976..9006)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9021..9042
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9159..9176)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9330..9918)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9410..9429)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(10089..10949)
/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 10712..10731
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10950..11054)
/gene="bla"
/label=AmpR promoter
primer_bind complement(11129..11148)
/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 agctcgttta gtgaaccgtc
601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
661 ccagcctccg gactctagag gatcgaaccc ttgccaccat ggacaagaag tactccattg
721 ggctcgctat cggcacaaac agcgtcggct gggccgtcat tacggacgag tacaaggtgc
781 cgagcaaaaa attcaaagtt ctgggcaata ccgatcgcca cagcataaag aagaacctca
841 ttggcgccct cctgttcgac tccggggaga cggccgaagc cacgcggctc aaaagaacag
901 cacggcgcag atatacccgc agaaagaatc ggatctgcta cctgcaggag atctttagta
961 atgagatggc taaggtggat gactctttct tccataggct ggaggagtcc tttttggtgg
1021 aggaggataa aaagcacgag cgccacccaa tctttggcaa tatcgtggac gaggtggcgt
1081 accatgaaaa gtacccaacc atatatcatc tgaggaagaa gcttgtagac agtactgata
1141 aggctgactt gcggttgatc tatctcgcgc tggcgcatat gatcaaattt cggggacact
1201 tcctcatcga gggggacctg aacccagaca acagcgatgt cgacaaactc tttatccaac
1261 tggttcagac ttacaatcag cttttcgaag agaacccgat caacgcatcc ggagttgacg
1321 ccaaagcaat cctgagcgct aggctgtcca aatcccggcg gctcgaaaac ctcatcgcac
1381 agctccctgg ggagaagaag aacggcctgt ttggtaatct tatcgccctg tcactcgggc
1441 tgacccccaa ctttaaatct aacttcgacc tggccgaaga tgccaagctt caactgagca
1501 aagacaccta cgatgatgat ctcgacaatc tgctggccca gatcggcgac cagtacgcag
1561 accttttttt ggcggcaaag aacctgtcag acgccattct gctgagtgat attctgcgag
1621 tgaacacgga gatcaccaaa gctccgctga gcgctagtat gatcaagcgc tatgatgagc
1681 accaccaaga cttgactttg ctgaaggccc ttgtcagaca gcaactgcct gagaagtaca
1741 aggaaatttt cttcgatcag tctaaaaatg gctacgccgg atacattgac ggcggagcaa
1801 gccaggagga attttacaaa tttattaagc ccatcttgga aaaaatggac ggcaccgagg
1861 agctgctggt aaagcttaac agagaagatc tgttgcgcaa acagcgcact ttcgacaatg
1921 gaagcatccc ccaccagatt cacctgggcg aactgcacgc tatcctcagg cggcaagagg
1981 atttctaccc ctttttgaaa gataacaggg aaaagattga gaaaatcctc acatttcgga
2041 taccctacta tgtaggcccc ctcgcccggg gaaattccag attcgcgtgg atgactcgca
2101 aatcagaaga gaccatcact ccctggaact tcgaggaagt cgtggataag ggggcctctg
2161 cccagtcctt catcgaaagg atgactaact ttgataaaaa tctgcctaac gaaaaggtgc
2221 ttcctaaaca ctctctgctg tacgagtact tcacagttta taacgagctc accaaggtca
2281 aatacgtcac agaagggatg agaaagccag cattcctgtc tggagagcag aagaaagcta
2341 tcgtggacct cctcttcaag acgaaccgga aagttaccgt gaaacagctc aaagaagact
2401 atttcaaaaa gattgaatgt ttcgactctg ttgaaatcag cggagtggag gatcgcttca
2461 acgcatccct gggaacgtat cacgatctcc tgaaaatcat taaagacaag gacttcctgg
2521 acaatgagga gaacgaggac attcttgagg acattgtcct cacccttacg ttgtttgaag
2581 atagggagat gattgaagaa cgcttgaaaa cttacgctca tctcttcgac gacaaagtca
2641 tgaaacagct caagaggcgc cgatatacag gatgggggcg gctgtcaaga aaactgatca
2701 atgggatccg agacaagcag agtggaaaga caatcctgga ttttcttaag tccgatggat
2761 ttgccaaccg gaacttcatg cagttgatcc atgatgactc tctcaccttt aaggaggaca
2821 tccagaaagc acaagtttct ggccaggggg acagtcttca cgagcacatc gctaatcttg
2881 caggtagccc agctatcaaa aagggaatac tgcagaccgt taaggtcgtg gatgaactcg
2941 tcaaagtaat gggaaggcat aagcccgaga atatcgttat cgagatggcc cgagagaacc
3001 aaactaccca gaagggacag aagaacagta gggaaaggat gaagaggatt gaagagggta
3061 taaaagaact ggggtcccaa atccttaagg aacacccagt tgaaaacacc cagcttcaga
3121 atgagaagct ctacctgtac tacctgcaga acggcaggga catgtacgtg gatcaggaac
3181 tggacatcaa tcggctctcc gactacgacg tggctgctat cgtgccccag tcttttctca
3241 aagatgattc tattgataat aaagtgttga caagatccga taaagctaga gggaagagtg
3301 ataacgtccc ctcagaagaa gttgtcaaga aaatgaaaaa ttattggcgg cagctgctga
3361 acgccaaact gatcacacaa cggaagttcg ataatctgac taaggctgaa cgaggtggcc
3421 tgtctgagtt ggataaagcc ggcttcatca aaaggcagct tgttgagaca cgccagatca
3481 ccaagcacgt ggcccaaatt ctcgattcac gcatgaacac caagtacgat gaaaatgaca
3541 aactgattcg agaggtgaaa gttattactc tgaagtctaa gctggtctca gatttcagaa
3601 aggactttca gttttataag gtgagagaga tcaacaatta ccaccatgcg catgatgcct
3661 acctgaatgc agtggtaggc actgcactta tcaaaaaata tcccaagctt gaatctgaat
3721 ttgtttacgg agactataaa gtgtacgatg ttaggaaaat gatcgcaaag tctgagcagg
3781 aaataggcaa ggccaccgct aagtacttct tttacagcaa tattatgaat tttttcaaga
3841 ccgagattac actggccaat ggagagattc ggaagcgacc acttatcgaa acaaacggag
3901 aaacaggaga aatcgtgtgg gacaagggta gggatttcgc gacagtccgg aaggtcctgt
3961 ccatgccgca ggtgaacatc gttaaaaaga ccgaagtaca gaccggaggc ttctccaagg
4021 aaagtatcct cccgaaaagg aacagcgaca agctgatcgc acgcaaaaaa gattgggacc
4081 ccaagaaata cggcggattc gattctccta cagtcgctta cagtgtactg gttgtggcca
4141 aagtggagaa agggaagtct aaaaaactca aaagcgtcaa ggaactgctg ggcatcacaa
4201 tcatggagcg atcaagcttc gaaaaaaacc ccatcgactt tctcgaggcg aaaggatata
4261 aagaggtcaa aaaagacctc atcattaagc ttcccaagta ctctctcttt gagcttgaaa
4321 acggccggaa acgaatgctc gctagtgcgg gcgagctgca gaaaggtaac gagctggcac
4381 tgccctctaa atacgttaat ttcttgtatc tggccagcca ctatgaaaag ctcaaagggt
4441 ctcccgaaga taatgagcag aagcagctgt tcgtggaaca acacaaacac taccttgatg
4501 agatcatcga gcaaataagc gaattctcca aaagagtgat cctcgccgac gctaacctcg
4561 ataaggtgct ttctgcttac aataagcaca gggataagcc catcagggag caggcagaaa
4621 acattatcca cttgtttact ctgaccaact tgggcgcgcc tgcagccttc aagtacttcg
4681 acaccaccat agacagaaag cggtacacct ctacaaagga ggtcctggac gccacactga
4741 ttcatcagtc aattacgggg ctctatgaaa caagaatcga cctctctcag ctcggtggag
4801 acagcagggc tgaccccaag aagaagagga aggtgtcgcc agggatccgt cgacttgacg
4861 cgttgatatc aacaagtttg tacaaaaaag caggctacaa agaggccagc ggttccggac
4921 gggctgacgc attggacgat tttgatctgg atatgctggg aagtgacgcc ctcgatgatt
4981 ttgaccttga catgcttggt tcggatgccc ttgatgactt tgacctcgac atgctcggca
5041 gtgacgccct tgatgatttc gacctggaca tgctgattaa ctctagaagt tccggatctc
5101 cgaaaaagaa acgcaaagtt ggtagccagt acctgcccga caccgacgac cggcaccgga
5161 tcgaggaaaa gcggaagcgg acctacgaga cattcaagag catcatgaag aagtccccct
5221 tcagcggccc caccgaccct agacctccac ctagaagaat cgccgtgccc agcagatcca
5281 gcgccagcgt gccaaaacct gccccccagc cttacccctt caccagcagc ctgagcacca
5341 tcaactacga cgagttccct accatggtgt tccccagcgg ccagatctct caggcctctg
5401 ctctggctcc agcccctcct caggtgctgc ctcaggctcc tgctcctgca ccagctccag
5461 ccatggtgtc tgcactggct caggcaccag cacccgtgcc tgtgctggct cctggacctc
5521 cacaggctgt ggctccacca gcccctaaac ctacacaggc cggcgagggc acactgtctg
5581 aagctctgct gcagctgcag ttcgacgacg aggatctggg agccctgctg ggaaacagca
5641 ccgatcctgc cgtgttcacc gacctggcca gcgtggacaa cagcgagttc cagcagctgc
5701 tgaaccaggg catccctgtg gcccctcaca ccaccgagcc catgctgatg gaataccccg
5761 aggccatcac ccggctcgtg acaggcgctc agaggcctcc tgatccagct cctgcccctc
5821 tgggagcacc aggcctgcct aatggactgc tgtctggcga cgaggacttc agctctatcg
5881 ccgatatgga tttctcagcc ttgctgggct ctggcagcgg cagccgggat tccagggaag
5941 ggatgttttt gccgaagcct gaggccggct ccgctattag tgacgtgttt gagggccgcg
6001 aggtgtgcca gccaaaacga atccggccat ttcatcctcc aggaagtcca tgggccaacc
6061 gcccactccc cgccagcctc gcaccaacac caaccggtcc agtacatgag ccagtcgggt
6121 cactgacccc ggcaccagtc cctcagccac tggatccagc gcccgcagtg actcccgagg
6181 ccagtcacct gttggaggat cccgatgaag agacgagcca ggctgtcaaa gcccttcggg
6241 agatggccga tactgtgatt ccccagaagg aagaggctgc aatctgtggc caaatggacc
6301 tttcccatcc gcccccaagg ggccatctgg atgagctgac aaccacactt gagtccatga
6361 ccgaggatct gaacctggac tcacccctga ccccggaatt gaacgagatt ctggatacct
6421 tcctgaacga cgagtgcctc ttgcatgcca tgcatatcag cacaggactg tccatcttcg
6481 acacatctct gttttgaaac ccagctttct tgtacaaagt ggttgatatc atctagagcg
6541 gccgcaggta cctgaataac taaggccgct tccctttagt gagggttaat gcttcgagca
6601 gacaaagggt tcgatcccta ccggttagta atgagtttaa acgggggagg ctaactgaaa
6661 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
6721 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
6781 tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc ttccttttcc
6841 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
6901 ggccctgcca tagcagatct gcgcagctgg ggctctaggg ggtatcccca cgcgccctgt
6961 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
7021 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
7081 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
7141 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
7201 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
7261 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
7321 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa
7381 ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
7441 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
7501 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
7561 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
7621 gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag ctattccaga
7681 agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg ggagcttgta
7741 tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg attgaacaag
7801 atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg
7861 cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc
7921 cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag gacgaggcag
7981 cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca
8041 ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat
8101 ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata
8161 cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac
8221 gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc
8281 tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc gaggatctcg
8341 tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg
8401 gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta
8461 cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg
8521 gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct
8581 gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct gccatcacga
8641 gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac
8701 gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccccaac
8761 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
8821 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
8881 catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc atagctgttt
8941 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
9001 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
9061 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
9121 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
9181 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
9241 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
9301 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
9361 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
9421 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
9481 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
9541 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
9601 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
9661 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
9721 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
9781 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
9841 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
9901 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
9961 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
10021 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
10081 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
10141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
10201 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
10261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
10321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
10381 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
10441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
10501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
10561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
10621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
10681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
10741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
10801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
10861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
10921 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
10981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
11041 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
11101 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
11161 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
11221 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
11281 ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
