| 质粒名称: | pLV-dCas9-p300-P2A-PuroR |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 83889 |
| 存储实验室: | Charles Gersbach |
|
载体骨架基本信息 |
|
| 载体名称: | lentiCRISPR v2 |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7800 bp |
| 完整质粒大小: | 14538 bp |
| 载体类型: | 哺乳动物表达，慢病毒，CRISPR |
| 筛选标记: | 嘌呤霉素 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | S.pyogenes dCas9 with c-terminal human p300 Core effector fusion (aa 1048-1664 of human p300) |
| 物种来源: | H. sapiens (human), Synthetic; S. pyogenes |
| 插入大小: | 6138 bp |
| 基因突变: | D10A; H840A in S.pyogenes Cas9 |
| GenBank编号 | WP_010922251.1 NP_001420.2 |
| 启动子 | EFS |
| 标签/融合蛋白: |
nucleoplasmin NLS（插入物上的C末端）
Flag （插接件上的C端子）
|
| 克隆方法: | Gibson克隆 |
| 5' 测序引物 | ATCCGGTGCCTAGAGAAGGT |
|
文献引用方法 |
|
| 材料和方法部分: | pLV-dCas9-p300-P2A-PuroR was a gift from Charles Gersbach (Addgene plasmid # 83889 ; http://n2t.net/addgene:83889 ; RRID:Addgene_83889) |
| 参考文献部分: | CRISPR-Cas9 epigenome editing enables high-throughput screening for functional regulatory elements in the human genome. Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB, Crawford GE, Reddy TE, Gersbach CA. Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853. 10.1038/nbt.3853 PubMed 28369033 |
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 pLV-dCas9-p300-P2A-PuroR 14538 bp ds-DNA circular SYN 18-MAY-2020
DEFINITION Lentiviral Sp dCas9-p300-P2A-PuroR.
ACCESSION .
VERSION .
KEYWORDS pLV-dCas9-p300-P2A-PuroR
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14538)
AUTHORS Klann TS, Black JB, Chellappan M, Safi A, Song L, Hilton IB,
Crawford GE, Reddy TE, Gersbach CA
TITLE CRISPR-Cas9 epigenome editing enables high-throughput screening for
functional regulatory elements in the human genome.
JOURNAL Nat Biotechnol. 2017 Apr 3. doi: 10.1038/nbt.3853.
PUBMED 28369033
REFERENCE 2 (bases 1 to 14538)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported May 18, 2020 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14538
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 1..380
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 382..580
/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"
LTR 598..778
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 825..950
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1443..1676
/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 2203..2320
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2440..2651
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 2670..2679
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2676..6779
/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 6780..6827
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6834..8684
/codon_start=1
/gene="EP300"
/product="catalytic histone acetyltransferase core of the
human E1A-associated protein p300 (Delvecchio et al.,
2013)"
/label=p300 core
/translation="IFKPEELRQALMPTLEALYRQDPESLPFRQPVDPQLLGIPDYFDI
VKSPMDLSTIKRKLDTGQYQEPWQYVDDIWLMFNNAWLYNRKTSRVYKYCSKLSEVFEQ
EIDPVMQSLGYCCGRKLEFSPQTLCCYGKQLCTIPRDATYYSYQNRYHFCEKCFNEIQG
ESVSLGDDPSQPQTTINKEQFSKRKNDTLDPELFVECTECGRKMHQICVLHHEIIWPAG
FVCDGCLKKSARTRKENKFSAKRLPSTRLGTFLENRVNDFLRRQNHPESGEVTVRVVHA
SDKTVEVKPGMKARFVDSGEMAESFPYRTKALFAFEEIDGVDLCFFGMHVQEYGSDCPP
PNQRRVYISYLDSVHFFRPKCLRTAVYHEILIGYLEYVKKLGYTTGHIWACPPSEGDDY
IFHCHPPDQKIPKPKRLQEWYKKMLDKAVSERIVHDYKDIFKQATEDRLTSAKELPYFE
GDFWPNVLEESIKELEQEEEERKREENTSNESTDVTKGDSKNAKKKNNKKTSKNKSSLS
RGNKKKPGMPNVSNDLSQKLYATMEKHKEVFFVIRLIAGPAANSLPPIVDPDPLIPCDL
MDGRDAFLTLARDKHLEFSSLRRAQWSTMCMLVELHTQSQD"
regulatory 8448..8457
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 8691..8714
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 8724..8780
/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 8781..9377
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="TEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERV
TELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAA
QQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSA
PRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 9274..9294
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 9393..9981
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(9446..9466)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(9864..9875)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 10053..10286
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(10312..10329)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 10318..10542
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 10588..11016
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(10675..10694)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 10885..10906
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(11025..11045)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 11030..11359
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 11210..11345
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 11272..11291
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 11407..11454
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 11473..11847
/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 11977..12098
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(12014..12033)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 12068..12087
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(12147..12163)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12147..12163)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(12160..12182)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 12171..12187
/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(12195..12225)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12240..12261
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(12378..12395)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(12549..13137)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(12629..12648)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(13308..14168)
/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 13931..13950
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(14169..14273)
/gene="bla"
/label=AmpR promoter
primer_bind complement(14348..14367)
/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 cgcgttttgc ctgtactggg
601 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
661 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
721 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
781 ggcgcccgaa cagggacttg aaagcgaaag ggaaaccaga ggagctctct cgacgcagga
841 ctcggcttgc tgaagcgcgc acggcaagag gcgaggggcg gcgactggtg agtacgccaa
901 aaattttgac tagcggaggc tagaaggaga gagatgggtg cgagagcgtc agtattaagc
961 gggggagaat tagatcgcga tgggaaaaaa ttcggttaag gccaggggga aagaaaaaat
1021 ataaattaaa acatatagta tgggcaagca gggagctaga acgattcgca gttaatcctg
1081 gcctgttaga aacatcagaa ggctgtagac aaatactggg acagctacaa ccatcccttc
1141 agacaggatc agaagaactt agatcattat ataatacagt agcaaccctc tattgtgtgc
1201 atcaaaggat agagataaaa gacaccaagg aagctttaga caagatagag gaagagcaaa
1261 acaaaagtaa gaccaccgca cagcaagcgg ccgctgatct tcagacctgg aggaggagat
1321 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
1381 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
1441 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
1501 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
1561 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
1621 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
1681 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
1741 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
1801 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
1861 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
1921 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
1981 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
2041 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
2101 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
2161 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
2221 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
2281 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
2341 cagagatcca gtttggttaa ttaaggtacc gaggcgatag ggttaaggga gctagctagg
2401 tcttgaaagg agtgggaatt ggctccggtg cccgtcagtg ggcagagcgc acatcgccca
2461 cagtccccga gaagttgggg ggaggggtcg gcaattgatc cggtgcctag agaaggtggc
2521 gcggggtaaa ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg
2581 gagaaccgta tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg
2641 ccagaacaca ggaccggttc tagagcgctg ccaccatgga caagaagtac agcatcggcc
2701 tggccatcgg caccaactct gtgggctggg ccgtgatcac cgacgagtac aaggtgccca
2761 gcaagaaatt caaggtgctg ggcaacaccg accggcacag catcaagaag aacctgatcg
2821 gagccctgct gttcgacagc ggcgaaacag ccgaggccac ccggctgaag agaaccgcca
2881 gaagaagata caccagacgg aagaaccgga tctgctatct gcaagagatc ttcagcaacg
2941 agatggccaa ggtggacgac agcttcttcc acagactgga agagtccttc ctggtggaag
3001 aggataagaa gcacgagcgg caccccatct tcggcaacat cgtggacgag gtggcctacc
3061 acgagaagta ccccaccatc taccacctga gaaagaaact ggtggacagc accgacaagg
3121 ccgacctgcg gctgatctat ctggccctgg cccacatgat caagttccgg ggccacttcc
3181 tgatcgaggg cgacctgaac cccgacaaca gcgacgtgga caagctgttc atccagctgg
3241 tgcagaccta caaccagctg ttcgaggaaa accccatcaa cgccagcggc gtggacgcca
3301 aggccatcct gtctgccaga ctgagcaaga gcagacggct ggaaaatctg atcgcccagc
3361 tgcccggcga gaagaagaat ggcctgttcg gaaacctgat tgccctgagc ctgggcctga
3421 cccccaactt caagagcaac ttcgacctgg ccgaggatgc caaactgcag ctgagcaagg
3481 acacctacga cgacgacctg gacaacctgc tggcccagat cggcgaccag tacgccgacc
3541 tgtttctggc cgccaagaac ctgtccgacg ccatcctgct gagcgacatc ctgagagtga
3601 acaccgagat caccaaggcc cccctgagcg cctctatgat caagagatac gacgagcacc
3661 accaggacct gaccctgctg aaagctctcg tgcggcagca gctgcctgag aagtacaaag
3721 agattttctt cgaccagagc aagaacggct acgccggcta cattgacggc ggagccagcc
3781 aggaagagtt ctacaagttc atcaagccca tcctggaaaa gatggacggc accgaggaac
3841 tgctcgtgaa gctgaacaga gaggacctgc tgcggaagca gcggaccttc gacaacggca
3901 gcatccccca ccagatccac ctgggagagc tgcacgccat tctgcggcgg caggaagatt
3961 tttacccatt cctgaaggac aaccgggaaa agatcgagaa gatcctgacc ttccgcatcc
4021 cctactacgt gggccctctg gccaggggaa acagcagatt cgcctggatg accagaaaga
4081 gcgaggaaac catcaccccc tggaacttcg aggaagtggt ggacaagggc gcttccgccc
4141 agagcttcat cgagcggatg accaacttcg ataagaacct gcccaacgag aaggtgctgc
4201 ccaagcacag cctgctgtac gagtacttca ccgtgtataa cgagctgacc aaagtgaaat
4261 acgtgaccga gggaatgaga aagcccgcct tcctgagcgg cgagcagaaa aaggccatcg
4321 tggacctgct gttcaagacc aaccggaaag tgaccgtgaa gcagctgaaa gaggactact
4381 tcaagaaaat cgagtgcttc gactccgtgg aaatctccgg cgtggaagat cggttcaacg
4441 cctccctggg cacataccac gatctgctga aaattatcaa ggacaaggac ttcctggaca
4501 atgaggaaaa cgaggacatt ctggaagata tcgtgctgac cctgacactg tttgaggaca
4561 gagagatgat cgaggaacgg ctgaaaacct atgcccacct gttcgacgac aaagtgatga
4621 agcagctgaa gcggcggaga tacaccggct ggggcaggct gagccggaag ctgatcaacg
4681 gcatccggga caagcagtcc ggcaagacaa tcctggattt cctgaagtcc gacggcttcg
4741 ccaacagaaa cttcatgcag ctgatccacg acgacagcct gacctttaaa gaggacatcc
4801 agaaagccca ggtgtccggc cagggcgata gcctgcacga gcacattgcc aatctggccg
4861 gcagccccgc cattaagaag ggcatcctgc agacagtgaa ggtggtggac gagctcgtga
4921 aagtgatggg ccggcacaag cccgagaaca tcgtgatcga aatggccaga gagaaccaga
4981 ccacccagaa gggacagaag aacagccgcg agagaatgaa gcggatcgaa gagggcatca
5041 aagagctggg cagccagatc ctgaaagaac accccgtgga aaacacccag ctgcagaacg
5101 agaagctgta cctgtactac ctgcagaatg ggcgggatat gtacgtggac caggaactgg
5161 acatcaaccg gctgtccgac tacgatgtgg acgccatcgt gcctcagagc tttctgaagg
5221 acgactccat cgacaacaag gtgctgacca gaagcgacaa gaaccggggc aagagcgaca
5281 acgtgccctc cgaagaggtc gtgaagaaga tgaagaacta ctggcggcag ctgctgaacg
5341 ccaagctgat tacccagaga aagttcgaca atctgaccaa ggccgagaga ggcggcctga
5401 gcgaactgga taaggccggc ttcatcaaga gacagctggt ggaaacccgg cagatcacaa
5461 agcacgtggc acagatcctg gactcccgga tgaacactaa gtacgacgag aatgacaagc
5521 tgatccggga agtgaaagtg atcaccctga agtccaagct ggtgtccgat ttccggaagg
5581 atttccagtt ttacaaagtg cgcgagatca acaactacca ccacgcccac gacgcctacc
5641 tgaacgccgt cgtgggaacc gccctgatca aaaagtaccc taagctggaa agcgagttcg
5701 tgtacggcga ctacaaggtg tacgacgtgc ggaagatgat cgccaagagc gagcaggaaa
5761 tcggcaaggc taccgccaag tacttcttct acagcaacat catgaacttt ttcaagaccg
5821 agattaccct ggccaacggc gagatccgga agcggcctct gatcgagaca aacggcgaaa
5881 ccggggagat cgtgtgggat aagggccggg attttgccac cgtgcggaaa gtgctgagca
5941 tgccccaagt gaatatcgtg aaaaagaccg aggtgcagac aggcggcttc agcaaagagt
6001 ctatcctgcc caagaggaac agcgataagc tgatcgccag aaagaaggac tgggacccta
6061 agaagtacgg cggcttcgac agccccaccg tggcctattc tgtgctggtg gtggccaaag
6121 tggaaaaggg caagtccaag aaactgaaga gtgtgaaaga gctgctgggg atcaccatca
6181 tggaaagaag cagcttcgag aagaatccca tcgactttct ggaagccaag ggctacaaag
6241 aagtgaaaaa ggacctgatc atcaagctgc ctaagtactc cctgttcgag ctggaaaacg
6301 gccggaagag aatgctggcc tctgccggcg aactgcagaa gggaaacgaa ctggccctgc
6361 cctccaaata tgtgaacttc ctgtacctgg ccagccacta tgagaagctg aagggctccc
6421 ccgaggataa tgagcagaaa cagctgtttg tggaacagca caagcactac ctggacgaga
6481 tcatcgagca gatcagcgag ttctccaaga gagtgatcct ggccgacgct aatctggaca
6541 aagtgctgtc cgcctacaac aagcaccggg ataagcccat cagagagcag gccgagaata
6601 tcatccacct gtttaccctg accaatctgg gagcccctgc cgccttcaag tactttgaca
6661 ccaccatcga ccggaagagg tacaccagca ccaaagaggt gctggacgcc accctgatcc
6721 accagagcat caccggcctg tacgagacac ggatcgacct gtctcagctg ggaggcgaca
6781 agcgacctgc cgccacaaag aaggctggac aggctaagaa gaagaaagga tccattttca
6841 aaccagaaga actacgacag gcactgatgc caactttgga ggcactttac cgtcaggatc
6901 cagaatccct tccctttcgt caacctgtgg accctcagct tttaggaatc cctgattact
6961 ttgatattgt gaagagcccc atggatcttt ctaccattaa gaggaagtta gacactggac
7021 agtatcagga gccctggcag tatgtcgatg atatttggct tatgttcaat aatgcctggt
7081 tatataaccg gaaaacatca cgggtataca aatactgctc caagctctct gaggtctttg
7141 aacaagaaat tgacccagtg atgcaaagcc ttggatactg ttgtggcaga aagttggagt
7201 tctctccaca gacactgtgt tgctacggca aacagttgtg cacaatacct cgtgatgcca
7261 cttattacag ttaccagaac aggtatcatt tctgtgagaa gtgtttcaat gagatccaag
7321 gggagagcgt ttctttgggg gatgaccctt cccagcctca aactacaata aataaagaac
7381 aattttccaa gagaaaaaat gacacactgg atcctgaact gtttgttgaa tgtacagagt
7441 gcggaagaaa gatgcatcag atctgtgtcc ttcaccatga gatcatctgg cctgctggat
7501 tcgtctgtga tggctgttta aagaaaagtg cacgaactag gaaagaaaat aagttttctg
7561 ctaaaaggtt gccatctacc agacttggca cctttctaga gaatcgtgtg aatgactttc
7621 tgaggcgaca gaatcaccct gagtcaggag aggtcactgt tagagtagtt catgcttctg
7681 acaaaaccgt ggaagtaaaa ccaggcatga aagcaaggtt tgtggacagt ggagagatgg
7741 cagaatcctt tccataccga accaaagccc tctttgcctt tgaagaaatt gatggtgttg
7801 acctgtgctt ctttggcatg catgttcaag agtatggctc tgactgccct ccacccaacc
7861 agaggagagt atacatatct tacctcgata gtgttcattt cttccgtcct aaatgcttga
7921 ggactgcagt ctatcatgaa atcctaattg gatatttaga atatgtcaag aaattaggtt
7981 acacaacagg gcatatttgg gcatgtccac caagtgaggg agatgattat atcttccatt
8041 gccatcctcc tgaccagaag atacccaagc ccaagcgact gcaggaatgg tacaaaaaaa
8101 tgcttgacaa ggctgtatca gagcgtattg tccatgacta caaggatatt tttaaacaag
8161 ctactgaaga tagattaaca agtgcaaagg aattgcctta tttcgagggt gatttctggc
8221 ccaatgttct ggaagaaagc attaaggaac tggaacagga ggaagaagag agaaaacgag
8281 aggaaaacac cagcaatgaa agcacagatg tgaccaaggg agacagcaaa aatgctaaaa
8341 agaagaataa taagaaaacc agcaaaaata agagcagcct gagtaggggc aacaagaaga
8401 aacccgggat gcccaatgta tctaacgacc tctcacagaa actatatgcc accatggaga
8461 agcataaaga ggtcttcttt gtgatccgcc tcattgctgg ccctgctgcc aactccctgc
8521 ctcccattgt tgatcctgat cctctcatcc cctgcgatct gatggatggt cgggatgcgt
8581 ttctcacgct ggcaagggac aagcacctgg agttctcttc actccgaaga gcccagtggt
8641 ccaccatgtg catgctggtg gagctgcaca cgcagagcca ggacggatcc gattacaaag
8701 acgatgacga taagggatcc ggcgcaacaa acttctctct gctgaaacaa gccggagatg
8761 tcgaagagaa tcctggaccg accgagtaca agcccacggt gcgcctcgcc acccgcgacg
8821 acgtccccag ggccgtacgc accctcgccg ccgcgttcgc cgactacccc gccacgcgcc
8881 acaccgtcga tccggaccgc cacatcgagc gggtcaccga gctgcaagaa ctcttcctca
8941 cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg
9001 tctggaccac gccggagagc gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca
9061 tggccgagtt gagcggttcc cggctggccg cgcagcaaca gatggaaggc ctcctggcgc
9121 cgcaccggcc caaggagccc gcgtggttcc tggccaccgt cggagtctcg cccgaccacc
9181 agggcaaggg tctgggcagc gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg
9241 gggtgcccgc cttcctggag acctccgcgc cccgcaacct ccccttctac gagcggctcg
9301 gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc
9361 gcaagcccgg tgcctgaacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
9421 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
9481 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
9541 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
9601 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
9661 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
9721 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
9781 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
9841 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
9901 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
9961 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
10021 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
10081 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
10141 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
10201 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
10261 ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg
10321 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
10381 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
10441 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
10501 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
10561 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
10621 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
10681 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
10741 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
10801 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
10861 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
10921 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
10981 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
11041 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
11101 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
11161 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
11221 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
11281 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
11341 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
11401 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
11461 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
11521 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
11581 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
11641 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
11701 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
11761 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
11821 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
11881 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
11941 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
12001 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
12061 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
12121 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
12181 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
12241 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
12301 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
12361 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
12421 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
12481 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
12541 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
12601 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
12661 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
12721 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
12781 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
12841 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
12901 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
12961 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
13021 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
13081 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
13141 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
13201 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
13261 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
13321 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
13381 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
13441 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
13501 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
13561 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
13621 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
13681 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
13741 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
13801 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
13861 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
13921 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
13981 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
14041 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
14101 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
14161 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
14221 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
14281 ccccgaaaag tgccacctga cgtcgacgga tcgggagatc tcccgatccc ctatggtgca
14341 ctctcagtac aatctgctct gatgccgcat agttaagcca gtatctgctc cctgcttgtg
14401 tgttggaggt cgctgagtag tgcgcgagca aaatttaagc tacaacaagg caaggcttga
14461 ccgacaattg catgaagaat ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac
14521 gggccagata tacgcgtt
//
