| 质粒名称: | pHR-SFFV-KRAB-dCas9-P2A-mCherry |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 60954 |
| 存储实验室: | Jonathan Weissman |
|
载体骨架基本信息 |
|
| 载体名称: | pHR |
| 载体出品公司: | Addgene |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 9000 bp |
| 完整质粒大小: | --- |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | KRAB-dCas9-P2A-mCherry fusion |
| 物种来源: | -- |
| 插入大小: | 5244 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | pHR-SFFV-KRAB-dCas9-P2A-mCherry was a gift from Jonathan Weissman (Addgene plasmid # 60954 ; http://n2t.net/addgene:60954 ; RRID:Addgene_60954) |
| 参考文献部分: | Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014 Oct 9. 10.1016/j.cell.2014.09.029 PubMed 25307932 |
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 pHR-SFFV-KRAB-dCas9-P2A-mCherry 14247 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION 2nd Generation Lentiviral vector. Expresses an N-terminal KRAB-dCas9
fusion protein and mCherry.
ACCESSION .
VERSION .
KEYWORDS pHR-SFFV-KRAB-dCas9-P2A-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14247)
AUTHORS Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead
EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann M,
Weissman JS
TITLE Genome-Scale CRISPR-Mediated Control of Gene Repression and
Activation.
JOURNAL Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029.
Epub 2014 Oct 9.
PUBMED 25307932
REFERENCE 2 (bases 1 to 14247)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14247
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(1..17)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(2..18)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 108..341
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
promoter complement(366..384)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(367..384)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(674..693)
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind complement(907..926)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 1026..1048
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind complement(1086..1104)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 1172..1276
/gene="bla"
/label=AmpR promoter
CDS 1277..2137
/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 complement(1495..1514)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 2308..2896
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 2797..2816
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 3050..3067
/label=L4440
/note="L4440 vector, forward primer"
primer_bind complement(3137..3157)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3142..3471
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3322..3457
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3384..3403
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
intron 4605..4670
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS 4800..4820
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
polyA_signal 5245..5379
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5282..5301)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 5336..5355
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
LTR 5547..6180
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 6227..6352
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 6849..7082
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature 7596..7713
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 7866..8273
/label=SFFV promoter
/note="spleen focus-forming virus long terminal repeat
(LTR) promoter"
regulatory 8348..8357
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 8384..8569
/codon_start=1
/gene="ZNF10"
/gene_synonym="KOX1"
/product="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
/label=KRAB
/translation="RTLVTFKDVFVDFTREEWKLLDTAQQIVYRNVMLENYKNLVSLGY
QLTKPDVILRLEKGEEP"
CDS 8591..12694
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=dCas9
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the D10A mutation in the RuvC
catalytic domain and the H840A mutation in the HNH
catalytic domain"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 12698..12724
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 12743..12763
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 12770..12790
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 12830..12886
/codon_start=1
/product="2A peptide from porcine teschovirus-1
polyprotein"
/label=P2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="ATNFSLLKQAGDVEENPGP"
CDS 12887..13597
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(13032..13050)
/label=mCherry-R
/note="mCherry, reverse primer"
primer_bind 13300..13319
/label=mCherry-F
/note="mCherry, forward primer"
misc_feature 13657..14245
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(13710..13730)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(14128..14139)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
ORIGIN
1 cgataccgtc gacctcgagg gaattaattc gagctcggta cctttaagac caatgactta
61 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
121 tcactcccaa cgaagacaag atctgctttt tgcttgtact gggtctctct ggttagacca
181 gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc ctcaataaag
241 cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag
301 atccctcaga cccttttagt cagtgtggaa aatctctagc agcatctaga attaattccg
361 tgtattctat agtgtcacct aaatcgtatg tgtatgatac ataaggttat gtattaattg
421 tagccgcgtt ctaacgacaa tatgtacaag cctaattgtg tagcatctgg cttactgaag
481 cagaccctat catctctctc gtaaactgcc gtcagagtcg gtttggttgg acgaaccttc
541 tgagtttctg gtaacgccgt cccgcacccg gaaatggtca gcgaaccaat cagcagggtc
601 atcgctagcc agatcctcta cgccggacgc atcgtggccg gcatcaccgg cgccacaggt
661 gcggttgctg gcgcctatat cgccgacatc accgatgggg aagatcgggc tcgccacttc
721 gggctcatga gcgcttgttt cggcgtgggt atggtggcag gccccgtggc cgggggactg
781 ttgggcgcca tctccttgca tgcaccattc cttgcggcgg cggtgctcaa cggcctcaac
841 ctactactgg gctgcttcct aatgcaggag tcgcataagg gagagcgtcg aatggtgcac
901 tctcagtaca atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc
961 cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac
1021 cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg
1081 aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta
1141 gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
1201 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
1261 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
1321 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
1381 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
1441 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
1501 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
1561 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
1621 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
1681 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
1741 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
1801 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
1861 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
1921 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
1981 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
2041 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
2101 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
2161 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
2221 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
2281 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
2341 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
2401 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct
2461 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
2521 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
2581 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
2641 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
2701 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
2761 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
2821 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
2881 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
2941 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
3001 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
3061 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
3121 tcattaatgc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
3181 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
3241 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
3301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
3361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
3421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagcttgga
3481 cacaagacag gcttgcgaga tatgtttgag aataccactt tatcccgcgt cagggagagg
3541 cagtgcgtaa aaagacgcgg actcatgtga aatactggtt tttagtgcgc cagatctcta
3601 taatctcgcg caacctattt tcccctcgaa cactttttaa gccgtagata aacaggctgg
3661 gacacttcac atgagcgaaa aatacatcgt cacctgggac atgttgcaga tccatgcacg
3721 taaactcgca agccgactga tgccttctga acaatggaaa ggcattattg ccgtaagccg
3781 tggcggtctg taccgggtgc gttactggcg cgtgaactgg gtattcgtca tgtcgatacc
3841 gtttgtattt ccagctacga tcacgacaac cagcgcgagc ttaaagtgct gaaacgcgca
3901 gaaggcgatg gcgaaggctt catcgttatt gatgacctgg tggataccgg tggtactgcg
3961 gttgcgattc gtgaaatgta tccaaaagcg cactttgtca ccatcttcgc aaaaccggct
4021 ggtcgtccgc tggttgatga ctatgttgtt gatatcccgc aagatacctg gattgaacag
4081 ccgtgggata tgggcgtcgt attcgtcccg ccaatctccg gtcgctaatc ttttcaacgc
4141 ctggcactgc cgggcgttgt tctttttaac ttcaggcggg ttacaatagt ttccagtaag
4201 tattctggag gctgcatcca tgacacaggc aaacctgagc gaaaccctgt tcaaaccccg
4261 ctttaaacat cctgaaacct cgacgctagt ccgccgcttt aatcacggcg cacaaccgcc
4321 tgtgcagtcg gcccttgatg gtaaaaccat ccctcactgg tatcgcatga ttaaccgtct
4381 gatgtggatc tggcgcggca ttgacccacg cgaaatcctc gacgtccagg cacgtattgt
4441 gatgagcgat gccgaacgta ccgacgatga tttatacgat acggtgattg gctaccgtgg
4501 cggcaactgg atttatgagt gggccccgga tctttgtgaa ggaaccttac ttctgtggtg
4561 tgacataatt ggacaaacta cctacagaga tttaaagctc taaggtaaat ataaaatttt
4621 taagtgtata atgtgttaaa ctactgattc taattgtttg tgtattttag attccaacct
4681 atggaactga tgaatgggag cagtggtgga atgcctttaa tgaggaaaac ctgttttgct
4741 cagaagaaat gccatctagt gatgatgagg ctactgctga ctctcaacat tctactcctc
4801 caaaaaagaa gagaaaggta gaagacccca aggactttcc ttcagaattg ctaagttttt
4861 tgagtcatgc tgtgtttagt aatagaactc ttgcttgctt tgctatttac accacaaagg
4921 aaaaagctgc actgctatac aagaaaatta tggaaaaata ttctgtaacc tttataagta
4981 ggcataacag ttataatcat aacatactgt tttttcttac tccacacagg catagagtgt
5041 ctgctattaa taactatgct caaaaattgt gtacctttag ctttttaatt tgtaaagggg
5101 ttaataagga atatttgatg tatagtgcct tgactagaga tcataatcag ccataccaca
5161 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
5221 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
5281 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
5341 ttgtccaaac tcatcaatgt atcttatcat gtctggatca actggataac tcaagctaac
5401 caaaatcatc ccaaacttcc caccccatac cctattacca ctgccaatta cctgtggttt
5461 catttactct aaacctgtga ttcctctgaa ttattttcat tttaaagaaa ttgtatttgt
5521 taaatatgta ctacaaactt agtagttgga agggctaatt cactcccaaa gaagacaaga
5581 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
5641 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
5701 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
5761 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
5821 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
5881 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
5941 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
6001 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
6061 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
6121 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
6181 gtggcgcccg aacagggact tgaaagcgaa agggaaacca gaggagctct ctcgacgcag
6241 gactcggctt gctgaagcgc gcacggcaag aggcgagggg cggcgactgg tgagtacgcc
6301 aaaaattttg actagcggag gctagaagga gagagatggg tgcgagagcg tcagtattaa
6361 gcgggggaga attagatcgc gatgggaaaa aattcggtta aggccagggg gaaagaaaaa
6421 atataaatta aaacatatag tatgggcaag cagggagcta gaacgattcg cagttaatcc
6481 tggcctgtta gaaacatcag aaggctgtag acaaatactg ggacagctac aaccatccct
6541 tcagacagga tcagaagaac ttagatcatt atataataca gtagcaaccc tctattgtgt
6601 gcatcaaagg atagagataa aagacaccaa ggaagcttta gacaagatag aggaagagca
6661 aaacaaaagt aagaccaccg cacagcaagc ggccggccgc tgatcttcag acctggagga
6721 ggagatatga gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa
6781 ccattaggag tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca
6841 gtgggaatag gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca
6901 gcgtcaatga cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag
6961 aacaatttgc tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc
7021 atcaagcagc tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc
7081 ctggggattt ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct
7141 agttggagta ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac
7201 agagaaatta acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag
7261 caagaaaaga atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg
7321 tttaacataa caaattggct gtggtatata aaattattca taatgatagt aggaggcttg
7381 gtaggtttaa gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat
7441 tcaccattat cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga
7501 atagaagaag aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatct
7561 cgacggtatc gccaaatggc agtattcatc cacaatttta aaagaaaagg ggggattggg
7621 gggtacagtg caggggaaag aatagtagac ataatagcaa cagacataca aactaaagaa
7681 ttacaaaaac aaattacaaa aattcaaaat tttcgggttt attacaggga cagcagagat
7741 ccagtttgga tcgataagct tgatatcgaa ttcctgcagc cccgataaaa taaaagattt
7801 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
7861 ctgcagtaac gccattttgc aaggcatgga aaaataccaa accaagaata gagaagttca
7921 gatcaagggc gggtacatga aaatagctaa cgttgggcca aacaggatat ctgcggtgag
7981 cagtttcggc cccggcccgg ggccaagaac agatggtcac cgcagtttcg gccccggccc
8041 gaggccaaga acagatggtc cccagatatg gcccaaccct cagcagtttc ttaagaccca
8101 tcagatgttt ccaggctccc ccaaggacct gaaatgaccc tgcgccttat ttgaattaac
8161 caatcagcct gcttctcgct tctgttcgcg cgcttctgct tcccgagctc tataaaagag
8221 ctcacaaccc ctcactcggc gcgccagtcc tccgacagac tgagtcgccc gggggggatc
8281 tggagctctc gagaattctc acgcgtctgc aggatatcaa gcttgcggta ccgcgggccc
8341 gggatccgcc accatggatg ctaagtcact aactgcctgg tcccggacac tggtgacctt
8401 caaggatgta tttgtggact tcaccaggga ggagtggaag ctgctggaca ctgctcagca
8461 gatcgtgtac agaaatgtga tgctggagaa ctataagaac ctggtttcct tgggttatca
8521 gcttactaag ccagatgtga tcctccggtt ggagaaggga gaagagcccg gcggttccgg
8581 cggagggtcg atggacaaga agtacagcat cggcctggcc atcggcacca actctgtggg
8641 ctgggccgtg atcaccgacg agtacaaggt gcccagcaag aaattcaagg tgctgggcaa
8701 caccgaccgg cacagcatca agaagaacct gatcggcgcc ctgctgttcg acagcggaga
8761 aacagccgag gccacccggc tgaagagaac cgccagaaga agatacacca gacggaagaa
8821 ccggatctgc tatctgcaag agatcttcag caacgagatg gccaaggtgg acgacagctt
8881 cttccacaga ctggaagagt ccttcctggt ggaagaggat aagaagcacg agcggcaccc
8941 catcttcggc aacatcgtgg acgaggtggc ctaccacgag aagtacccca ccatctacca
9001 cctgagaaag aaactggtgg acagcaccga caaggccgac ctgcggctga tctatctggc
9061 cctggcccac atgatcaagt tccggggcca cttcctgatc gagggcgacc tgaaccccga
9121 caacagcgac gtggacaagc tgttcatcca gctggtgcag acctacaacc agctgttcga
9181 ggaaaacccc atcaacgcca gcggcgtgga cgccaaggcc atcctgtctg ccagactgag
9241 caagagcaga cggctggaaa atctgatcgc ccagctgccc ggcgagaaga agaatggcct
9301 gttcggcaac ctgattgccc tgagcctggg cctgaccccc aacttcaaga gcaacttcga
9361 cctggccgag gatgccaaac tgcagctgag caaggacacc tacgacgacg acctggacaa
9421 cctgctggcc cagatcggcg accagtacgc cgacctgttt ctggccgcca agaacctgtc
9481 cgacgccatc ctgctgagcg acatcctgag agtgaacacc gagatcacca aggcccccct
9541 gagcgcctct atgatcaaga gatacgacga gcaccaccag gacctgaccc tgctgaaagc
9601 tctcgtgcgg cagcagctgc ctgagaagta caaagagatt ttcttcgacc agagcaagaa
9661 cggctacgcc ggctacatcg atggcggagc cagccaggaa gagttctaca agttcatcaa
9721 gcccatcctg gaaaagatgg acggcaccga ggaactgctc gtgaagctga acagagagga
9781 cctgctgcgg aagcagcgga ccttcgacaa cggcagcatc ccccaccaga tccacctggg
9841 agagctgcac gccattctgc ggcggcagga agatttttac ccattcctga aggacaaccg
9901 ggaaaagatc gagaagatcc tgaccttccg catcccctac tacgtgggcc ctctggccag
9961 gggaaacagc agattcgcct ggatgaccag aaagagcgag gaaaccatca ccccctggaa
10021 cttcgaggaa gtggtggaca agggcgccag cgcccagagc ttcatcgagc ggatgaccaa
10081 cttcgataag aacctgccca acgagaaggt gctgcccaag cacagcctgc tgtacgagta
10141 cttcaccgtg tacaacgagc tgaccaaagt gaaatacgtg accgagggaa tgagaaagcc
10201 cgccttcctg agcggcgagc agaaaaaagc catcgtggac ctgctgttca agaccaaccg
10261 gaaagtgacc gtgaagcagc tgaaagagga ctacttcaag aaaatcgagt gcttcgactc
10321 cgtggaaatc tccggcgtgg aagatcggtt caacgcctcc ctgggcacat accacgatct
10381 gctgaaaatt atcaaggaca aggacttcct ggacaatgag gaaaacgagg acattctgga
10441 agatatcgtg ctgaccctga cactgtttga ggacagagag atgatcgagg aacggctgaa
10501 aacctatgcc cacctgttcg acgacaaagt gatgaagcag ctgaagcggc ggagatacac
10561 cggctggggc aggctgagcc ggaagctgat caacggcatc cgggacaagc agtccggcaa
10621 gacaatcctg gatttcctga agtccgacgg cttcgccaac agaaacttca tgcagctgat
10681 ccacgacgac agcctgacct ttaaagagga catccagaaa gcccaggtgt ccggccaggg
10741 cgatagcctg cacgagcaca ttgccaatct ggccggcagc cccgccatta agaagggcat
10801 cctgcagaca gtgaaggtgg tggacgagct cgtgaaagtg atgggccggc acaagcccga
10861 gaacatcgtg atcgaaatgg ccagagagaa ccagaccacc cagaagggac agaagaacag
10921 ccgcgagaga atgaagcgga tcgaagaggg catcaaagag ctgggcagcc agatcctgaa
10981 agaacacccc gtggaaaaca cccagctgca gaacgagaag ctgtacctgt actacctgca
11041 gaatgggcgg gatatgtacg tggaccagga actggacatc aaccggctgt ccgactacga
11101 tgtggacgct atcgtgcctc agagctttct gaaggacgac tccatcgata acaaagtgct
11161 gactcggagc gacaagaacc ggggcaagag cgacaacgtg ccctccgaag aggtcgtgaa
11221 gaagatgaag aactactggc gccagctgct gaatgccaag ctgattaccc agaggaagtt
11281 cgacaatctg accaaggccg agagaggcgg cctgagcgaa ctggataagg ccggcttcat
11341 caagagacag ctggtggaaa cccggcagat cacaaagcac gtggcacaga tcctggactc
11401 ccggatgaac actaagtacg acgagaacga caaactgatc cgggaagtga aagtgatcac
11461 cctgaagtcc aagctggtgt ccgatttccg gaaggatttc cagttttaca aagtgcgcga
11521 gatcaacaac taccaccacg cccacgacgc ctacctgaac gccgtcgtgg gaaccgccct
11581 gatcaaaaag taccctaagc tggaaagcga gttcgtgtac ggcgactaca aggtgtacga
11641 cgtgcggaag atgatcgcca agagcgagca ggaaatcggc aaggctaccg ccaagtactt
11701 cttctacagc aacatcatga actttttcaa gaccgagatt accctggcca acggcgagat
11761 ccggaagcgg cctctgatcg agacaaacgg cgaaacaggc gagatcgtgt gggataaggg
11821 ccgggacttt gccaccgtgc ggaaagtgct gtctatgccc caagtgaata tcgtgaaaaa
11881 gaccgaggtg cagacaggcg gcttcagcaa agagtctatc ctgcccaaga ggaacagcga
11941 caagctgatc gccagaaaga aggactggga ccctaagaag tacggcggct tcgacagccc
12001 caccgtggcc tattctgtgc tggtggtggc caaagtggaa aagggcaagt ccaagaaact
12061 gaagagtgtg aaagagctgc tggggatcac catcatggaa agaagcagct tcgagaagaa
12121 tcccatcgac tttctggaag ccaagggcta caaagaagtg aaaaaggacc tgatcatcaa
12181 gctgcctaag tactccctgt tcgagctgga aaacggccgg aagagaatgc tggcctctgc
12241 cggcgaactg cagaagggaa acgaactggc cctgccctcc aaatatgtga acttcctgta
12301 cctggccagc cactatgaga agctgaaggg ctcccccgag gataatgagc agaaacagct
12361 gtttgtggaa cagcacaaac actacctgga cgagatcatc gagcagatca gcgagttctc
12421 caagagagtg atcctggccg acgctaatct ggacaaggtg ctgagcgcct acaacaagca
12481 cagagacaag cctatcagag agcaggccga gaatatcatc cacctgttta ccctgaccaa
12541 tctgggagcc cctgccgcct tcaagtactt tgacaccacc atcgaccgga agaggtacac
12601 cagcaccaaa gaggtgctgg acgccaccct gatccaccag agcatcaccg gcctgtacga
12661 gacacggatc gacctgtctc agctgggagg cgacgcctat ccctatgacg tgcccgatta
12721 tgccagcctg ggcagcggct cccccaagaa aaaacgcaag gtggaagatc ctaagaaaaa
12781 gcggaaagtg gacggcattg gtagtgggag caacggcagc agcggatccg ctactaactt
12841 cagcctgctg aagcaggctg gagacgtgga ggagaaccct ggacctatgg tgagcaaggg
12901 cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc acatggaggg
12961 ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc cctacgaggg
13021 cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg cctgggacat
13081 cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg ccgacatccc
13141 cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga tgaacttcga
13201 ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg agttcatcta
13261 caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc agaagaagac
13321 catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc tgaagggcga
13381 gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg tcaagaccac
13441 ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca tcaagttgga
13501 catcacctcc cacaacgagg actacaccat cgtggaacag tacgaacgcg ccgagggccg
13561 ccactccacc ggcggcatgg acgagctgta caagtaagcg gccgcgactc tagagtcgac
13621 ctgcaggcat gcaagcttga tatcaagctt atcgataatc aacctctgga ttacaaaatt
13681 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
13741 gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt ctcctccttg
13801 tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag gcaacgtggc
13861 gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc caccacctgt
13921 cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga actcatcgcc
13981 gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa ttccgtggtg
14041 ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac ctggattctg
14101 cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct tccttcccgc
14161 ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca gacgagtcgg
14221 atctcccttt gggccgcctc cccgcat
//
