| 质粒名称: | Lenti-dCas9-KRAB-blast |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 89567 |
| 存储实验室: | Gary Hon |
|
载体骨架基本信息 |
|
| 载体名称: | plenti |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 14229 bp |
| 完整质粒大小: | -- |
| 载体修饰: | 用KRAB域替换VP64 |
| 载体类型: | 慢病毒载体 |
| 筛选标记: | 杀稻瘟菌素 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9 |
| 物种来源: | Synthetic |
| 插入大小: | -- |
| 基因突变: | D10A and H840A |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: |
KRAB（插入件上的C端子）
Blasticidin resistance gene（插入端C端）
|
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | Lenti-dCas9-KRAB-blast was a gift from Gary Hon (Addgene plasmid # 89567 ; http://n2t.net/addgene:89567 ; RRID:Addgene_89567) |
| 参考文献部分: | Multiplexed Engineering and Analysis of Combinatorial Enhancer Activity in Single Cells. Xie S, Duan J, Li B, Zhou P, Hon GC. Mol Cell. 2017 Apr 20;66(2):285-299.e5. doi: 10.1016/j.molcel.2017.03.007. Epub 2017 Apr 13. 10.1016/j.molcel.2017.03.007 PubMed 28416141 |
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 Lenti-dCas9-KRAB-blast 14229 bp ds-DNA circular SYN 22-APR-2019
DEFINITION Plasmid expression dCas9 protein in fusion with KRAB domain.
ACCESSION .
VERSION .
KEYWORDS Lenti-dCas9-KRAB-blast
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14229)
AUTHORS Xie S, Duan J, Li B, Zhou P, Hon GC
TITLE Multiplexed Engineering and Analysis of Combinatorial Enhancer
Activity in Single Cells.
JOURNAL Mol Cell. 2017 Apr 20;66(2):285-299.e5. doi:
10.1016/j.molcel.2017.03.007. Epub 2017 Apr 13.
PUBMED 28416141
REFERENCE 2 (bases 1 to 14229)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 22, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14229
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(47..66)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 238..617
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 619..817
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 772..792
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 835..1015
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1062..1187
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1680..1913
/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 2440..2557
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2687..3865
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 2918..3856
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 3813..3833
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 3881..3928
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 3929..8029
/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 N863A mutation in the HNH
catalytic domain"
/translation="DKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKARGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 8084..8104
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8147..8341
/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="RTLVTFKDVFVDFTREEWKLLDTAQQILYRNVMLENYKNLVSLGY
QLTKPDVILRLEKGEEPWLV"
CDS 8408..8428
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 8444..8497
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 8498..8896
/codon_start=1
/gene="Aspergillus terreus BSD"
/product="blasticidin S deaminase"
/label=BSD
/note="confers resistance to blasticidin"
/translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
KAIVKDSDGQPTAVGIRELLPSGYVWEG"
misc_feature 8921..9509
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(8974..8994)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9392..9403)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(9512..9528)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9513..9529)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 10034..10214
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(10240..10257)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 10246..10470
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 10516..10944
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(10603..10622)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10813..10834
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(10953..10973)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 10958..11287
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 11138..11273
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 11200..11219
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 11335..11382
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 11401..11775
/codon_start=1
/gene="Sh ble from Streptoalloteichus hindustanus"
/product="antibiotic-binding protein"
/label=BleoR
/note="confers resistance to bleomycin, phleomycin, and
Zeocin(TM)"
/translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
EFALRDPAGNCVHFVAEEQD"
polyA_signal 11905..12026
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(11942..11961)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 11996..12015
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(12075..12091)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12075..12091)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(12088..12110)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 12099..12115
/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(12123..12153)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12168..12189
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(12306..12323)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(12477..13065)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(12557..12576)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(13236..14096)
/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 13859..13878
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(14097..14201)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
241 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
301 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
361 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
421 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
481 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
541 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
601 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
661 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
721 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
781 gcggtaggcg tgtacggtgg gaggtctata taagcagcgc gttttgcctg tactgggtct
841 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
901 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
961 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtggc
1021 gcccgaacag ggacttgaaa gcgaaaggga aaccagagga gctctctcga cgcaggactc
1081 ggcttgctga agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa
1141 ttttgactag cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg
1201 ggagaattag atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata
1261 aattaaaaca tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc
1321 tgttagaaac atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga
1381 caggatcaga agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc
1441 aaaggataga gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca
1501 aaagtaagac caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg
1561 agggacaatt ggagaagtga attatataaa tataaagtag taaaaattga accattagga
1621 gtagcaccca ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata
1681 ggagctttgt tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcgtcaatg
1741 acgctgacgg tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg
1801 ctgagggcta ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag
1861 ctccaggcaa gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt
1921 tggggttgct ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt
1981 aataaatctc tggaacagat ttggaatcac acgacctgga tggagtggga cagagaaatt
2041 aacaattaca caagcttaat acactcctta attgaagaat cgcaaaacca gcaagaaaag
2101 aatgaacaag aattattgga attagataaa tgggcaagtt tgtggaattg gtttaacata
2161 acaaattggc tgtggtatat aaaattattc ataatgatag taggaggctt ggtaggttta
2221 agaatagttt ttgctgtact ttctatagtg aatagagtta ggcagggata ttcaccatta
2281 tcgtttcaga cccacctccc aaccccgagg ggacccgaca ggcccgaagg aatagaagaa
2341 gaaggtggag agagagacag agacagatcc attcgattag tgaacggatc ggcactgcgt
2401 gcgccaattc tgcagacaaa tggcagtatt catccacaat tttaaaagaa aaggggggat
2461 tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca tacaaactaa
2521 agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca gggacagcag
2581 agatccagtt tggttaatta gctagctgca aagatggata aagttttaaa cagagaggaa
2641 tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct ccggtgcccg
2701 tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa
2761 ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg
2821 gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa
2881 cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt gtggttcccg
2941 cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt ccacctggct
3001 gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc
3061 ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg
3121 ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg ataagtctct
3181 agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag atagtcttgt
3241 aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg gcggcgacgg
3301 ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc ggccaccgag
3361 aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc
3421 gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga
3481 aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg
3541 agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc
3601 ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt tctcgagctt
3661 ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt ttccccacac
3721 tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct ccttggaatt
3781 tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg ttcaaagttt
3841 ttttcttcca tttcaggtgt cgtgacgtac ggccaccatg aaaaggccgg cggccacgaa
3901 aaaggccggc caggcaaaaa agaaaaagga caagaagtac agcatcggcc tggccatcgg
3961 caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca gcaagaaatt
4021 caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg gagccctgct
4081 gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca gaagaagata
4141 caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg agatggccaa
4201 ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag aggataagaa
4261 gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc acgagaagta
4321 ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg ccgacctgcg
4381 gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc tgatcgaggg
4441 cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg tgcagaccta
4501 caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca aggccatcct
4561 gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc tgcccggcga
4621 gaagaagaat ggcctgttcg gcaacctgat tgccctgagc ctgggcctga cccccaactt
4681 caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg acacctacga
4741 cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc tgtttctggc
4801 cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga acaccgagat
4861 caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc accaggacct
4921 gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag agattttctt
4981 cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc aggaagagtt
5041 ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac tgctcgtgaa
5101 gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca gcatccccca
5161 ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt tttacccatt
5221 cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc cctactacgt
5281 gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga gcgaggaaac
5341 catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc agagcttcat
5401 cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc ccaagcacag
5461 cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat acgtgaccga
5521 gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg tggacctgct
5581 gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact tcaagaaaat
5641 cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg cctccctggg
5701 cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca atgaggaaaa
5761 cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca gagagatgat
5821 cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga agcagctgaa
5881 gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg gcatccggga
5941 caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg ccaacagaaa
6001 cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc agaaagccca
6061 ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg gcagccccgc
6121 cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga aagtgatggg
6181 ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga ccacccagaa
6241 gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca aagagctggg
6301 cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg agaagctgta
6361 cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg acatcaaccg
6421 gctgtccgac tacgatgtgg accacatcgt gcctcagagc tttctgaagg acgactccat
6481 cgacaacaag gtgctgacca gaagcgacaa ggcccggggc aagagcgaca acgtgccctc
6541 cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg ccaagctgat
6601 tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga gcgaactgga
6661 taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa agcacgtggc
6721 acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc tgatccggga
6781 agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg atttccagtt
6841 ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc tgaacgccgt
6901 cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg tgtacggcga
6961 ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa tcggcaaggc
7021 taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg agattaccct
7081 ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa ccggggagat
7141 cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca tgccccaagt
7201 gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt ctatcctgcc
7261 caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta agaagtacgg
7321 cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag tggaaaaggg
7381 caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca tggaaagaag
7441 cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag aagtgaaaaa
7501 ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg gccggaagag
7561 aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc cctccaaata
7621 tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc ccgaggataa
7681 tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga tcatcgagca
7741 gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca aagtgctgtc
7801 cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata tcatccacct
7861 gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca ccaccatcga
7921 ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc accagagcat
7981 caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca gcgctggagg
8041 aggtggaagc ggaggaggag gaagcggagg aggaggtagc ggacctaaga aaaagaggaa
8101 ggtggcggcc gctggatccg atgctaagtc actgactgcc tggtcccgga cactggtgac
8161 cttcaaggat gtgtttgtgg acttcaccag ggaggagtgg aagctgctgg acactgctca
8221 gcagatcctg tacagaaatg tgatgctgga gaactataag aacctggttt ccttgggtta
8281 tcagcttact aagccagatg tgatcctccg gttggagaag ggagaagagc cctggctggt
8341 ggagagagaa attcaccaag agacccatcc tgattcagag actgcatttg aaatcaaatc
8401 atcagttccg aaaaagaaac gcaaagtttg tacaggcagt ggagagggca gaggaagtct
8461 gctaacatgc ggtgacgtcg aggagaatcc tggcccaatg gccaagcctt tgtctcaaga
8521 agaatccacc ctcattgaaa gagcaacggc tacaatcaac agcatcccca tctctgaaga
8581 ctacagcgtc gccagcgcag ctctctctag cgacggccgc atcttcactg gtgtcaatgt
8641 atatcatttt actgggggac cttgtgcaga actcgtggtg ctgggcactg ctgctgctgc
8701 ggcagctggc aacctgactt gtatcgtcgc gatcggaaat gagaacaggg gcatcttgag
8761 cccctgcgga cggtgccgac aggtgcttct cgatctgcat cctgggatca aagccatagt
8821 gaaggacagt gatggacagc cgacggcagt tgggattcgt gaattgctgc cctctggtta
8881 tgtgtgggag ggctaagaat tcgatatcaa gcttatcggt aatcaacctc tggattacaa
8941 aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata
9001 cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc
9061 cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg
9121 tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac
9181 ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat
9241 cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt
9301 ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat
9361 tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc
9421 ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag
9481 tcggatctcc ctttgggccg cctccccgca tcgataccgt cgacctcgag acctagaaaa
9541 acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga
9601 agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat
9661 gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaaggggg gactggaagg
9721 gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg
9781 ctacttccct gattggcaga actacacacc agggccaggg atcagatatc cactgacctt
9841 tggatggtgc tacaagctag taccagttga gcaagagaag gtagaagaag ccaatgaagg
9901 agagaacacc cgcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga
9961 agtattagag tggaggtttg acagccgcct agcatttcat cacatggccc gagagctgca
10021 tccggactgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
10081 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
10141 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
10201 gaaaatctct agcagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt
10261 gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc
10321 ccactgtcct ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt
10381 ctattctggg gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca
10441 ggcatgctgg ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct
10501 ctagggggta tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
10561 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
10621 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
10681 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
10741 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
10801 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
10861 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
10921 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
10981 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
11041 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
11101 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
11161 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
11221 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
11281 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
11341 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
11401 atggccaagt tgaccagtgc cgttccggtg ctcaccgcgc gcgacgtcgc cggagcggtc
11461 gagttctgga ccgaccggct cgggttctcc cgggacttcg tggaggacga cttcgccggt
11521 gtggtccggg acgacgtgac cctgttcatc agcgcggtcc aggaccaggt ggtgccggac
11581 aacaccctgg cctgggtgtg ggtgcgcggc ctggacgagc tgtacgccga gtggtcggag
11641 gtcgtgtcca cgaacttccg ggacgcctcc gggccggcca tgaccgagat cggcgagcag
11701 ccgtgggggc gggagttcgc cctgcgcgac ccggccggca actgcgtgca cttcgtggcc
11761 gaggagcagg actgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg
11821 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
11881 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
11941 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
12001 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
12061 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
12121 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
12181 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
12241 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
12301 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
12361 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
12421 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
12481 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
12541 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
12601 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
12661 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
12721 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
12781 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
12841 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
12901 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
12961 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
13021 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
13081 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
13141 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
13201 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
13261 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
13321 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
13381 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
13441 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
13501 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
13561 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
13621 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
13681 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
13741 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
13801 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
13861 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
13921 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
13981 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
14041 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
14101 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
14161 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
14221 ccacctgac
//
