| 质粒名称: | lenti-SpCas9 puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 104994 |
| 存储实验室: | Brett Stringer |
|
载体骨架基本信息 |
|
| 载体名称: | lentiCRISPRv2 puro (Addgene #98290) |
| 载体出品公司: | Brett Stringer from Feng Zhang |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 12671bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 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: | KpnI-XhoI-BsrGI-NheI |
| 物种来源: | -- |
| 插入大小: | 24bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | 人细胞色素c氧化酶亚基VIII（N端） |
| 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: | lenti-SpCas9 puro was a gift from Brett Stringer (Addgene plasmid # 104994 ; http://n2t.net/addgene:104994 ; RRID:Addgene_104994) |
| 参考文献部分: | A reference collection of patient-derived cell line and xenograft models of proneural, classical and mesenchymal glioblastoma. Stringer BW, Day BW, D'Souza RCJ, Jamieson PR, Ensbey KS, Bruce ZC, Lim YC, Goasdoue K, Offenhauser C, Akgul S, Allan S, Robertson T, Lucas P, Tollesson G, Campbell S, Winter C, Do H, Dobrovic A, Inglis PL, Jeffree RL, Johns TG, Boyd AW. Sci Rep. 2019 Mar 20;9(1):4902. doi: 10.1038/s41598-019-41277-z. 10.1038/s41598-019-41277-z PubMed 30894629 |
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 lentiCRISPRv2 puro 14877 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Variant of lentiCRISPRv2 that confers puromycin resistance.
ACCESSION .
VERSION .
KEYWORDS lentiCRISPRv2 puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14877)
AUTHORS Brett Stringer
TITLE Brett Stringer Lab Plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 14877)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14877
/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 242..621
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 623..821
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 776..796
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 839..1019
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1066..1191
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1684..1917
/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 2444..2561
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2612..2852
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind 2612..2632
/label=hU6-F
/note="Human U6 promoter, forward primer"
primer_bind 2783..2802
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
misc_RNA 4742..4817
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter 4879..5090
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 5109..5118
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 5115..9218
/codon_start=1
/product="Cas9 (Csn1) endonuclease from the Streptococcus
pyogenes Type II CRISPR/Cas system"
/label=Cas9
/note="generates RNA-guided double strand breaks in DNA"
/translation="MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
CDS 9219..9266
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 9267..9290
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 9300..9356
/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 9357..9953
/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 9850..9870
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 9969..10557
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(10022..10042)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(10440..10451)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 10629..10862
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(10888..10905)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 10894..11118
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 11164..11592
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11251..11270)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11461..11482
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(11601..11621)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 11606..11935
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 11786..11921
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 11848..11867
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 11983..12030
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 12049..12423
/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 12553..12674
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(12590..12609)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 12644..12663
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(12723..12739)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(12723..12739)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(12736..12758)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 12747..12763
/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(12771..12801)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 12816..12837
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(12954..12971)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(13125..13713)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(13205..13224)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(13884..14744)
/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 14507..14526
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(14745..14849)
/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 acgcgcgcgt
241 tgacattgat tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc
301 ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc
361 aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg
421 actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat
481 caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc
541 tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta catctacgta
601 ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag
661 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
721 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
781 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gcgcgttttg cctgtactgg
841 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
901 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
961 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
1021 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
1081 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
1141 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
1201 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
1261 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
1321 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
1381 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
1441 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
1501 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
1561 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
1621 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
1681 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
1741 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
1801 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
1861 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
1921 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
1981 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
2041 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
2101 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
2161 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
2221 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
2281 attatcgttt cagacccacc tcccaacccc gaggggaccc gacaggcccg aaggaataga
2341 agaagaaggt ggagagagag acagagacag atccattcga ttagtgaacg gatcggcact
2401 gcgtgcgcca attctgcaga caaatggcag tattcatcca caattttaaa agaaaagggg
2461 ggattggggg gtacagtgca ggggaaagaa tagtagacat aatagcaaca gacatacaaa
2521 ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt tcgggtttat tacagggaca
2581 gcagagatcc agtttggtta attaaggtac cgagggccta tttcccatga ttccttcata
2641 tttgcatata cgatacaagg ctgttagaga gataattaga attaatttga ctgtaaacac
2701 aaagatatta gtacaaaata cgtgacgtag aaagtaataa tttcttgggt agtttgcagt
2761 tttaaaatta tgttttaaaa tggactatca tatgcttacc gtaacttgaa agtatttcga
2821 tttcttggct ttatatatct tgtggaaagg acgaaacacc ggagacggtt gtaaatgagc
2881 acacaaaata cacatgctaa aatattatat tctatgacct ttataaaatc aaccaaaatc
2941 ttctttttaa taactttagt atcaataatt agaattttta tgttcctttt tgcaaacttt
3001 taataaaaat gagcaaaata aaaaaacgct agttttagta actcgcgttg ttttcttcac
3061 ctttaataat agctactcca ccacttgttc ctaagcggtc agctcctgct tcaatcattt
3121 tttgagcatc ttcaaatgtt ctaactccac cagctgcttt aactaaagca ttgtctttaa
3181 caactgactt cattagttta acatcttcaa atgttgcacc tgattttgaa aatcctgttg
3241 atgttttaac aaattctaat ccagcttcaa cagctatttc acaagctttc atgatttctt
3301 cttttgttaa taaacaattt tccataatac atttaacaac atgtgatcca gctgcttttt
3361 ttacagcttt catgtcttct aaaactaatt cataattttt gtcttttaat gcaccaatat
3421 ttaataccat atcaatttct gttgcaccat ctttaattgc ttcagaaact tcgaatgctt
3481 ttgtagctgt tgtgcatgca cctagaggaa aacctacaac atttgttatt cctacatttg
3541 tgccttttaa taattcttta caatagcttg ttcaatatga attaacacaa actgttgcaa
3601 aatcaaattc aattgcttca tcacataatt gtttaatttc agctttcgta gcatcttgtt
3661 ttaataatgt gtgatctata tatttgttta gtttcatttt ttctcctata tattcatttt
3721 taattttaat tctttaataa tttcgtctac tttaacttta gcgttttgaa cagattcacc
3781 aacacctata aaataaattt ttagtttagg ttcagttcca cttgggcgaa cagcaaatca
3841 tgacttatct tctaaataaa attttagtaa gtcttgtcct ggcatattat acattccatc
3901 gatgtagtct tcaacattaa caactttaag tccagcaatt tgagttaagg gtgttgctct
3961 caatgatttc attaatggtt caatttttaa tttcttttct tctggtttaa aattcaagtt
4021 taaagtgaaa gtgtaatatg cacccatttc tttaaataaa tcttctaaat agtctactaa
4081 tgttttattt tgttttttat aaaatcaagc agcctctgct attaatatag aagcttgtat
4141 tccatcttta tctctagctg agtcatcaat tacatatcca taactttctt cataagcaaa
4201 aacaaaattt aatccgttat cttcttcttt agcaatttct ctacccattc atttaaatcc
4261 agttaaagtt tttacaatat taactccata tttttcatga gcgattctat cacccaaatc
4321 acttgttaca aaacttgaat atagagccgg attttttgga atgctattta agcgttttag
4381 atttgataat tttcaatcaa ttaaaattgg tcctgtttga tttccatcta atcttacaaa
4441 atgaccatca tgttttattg ccattccaaa tctgtcagca tctgggtcat tcataataat
4501 aatatctgca tcatgtttaa taccatattc aagcggtatt tttcatgcag gatcaaattc
4561 tggatttgga tttacaacat ttttaaatgt ttcatcttca aatgcatgct cttcaacctc
4621 aataacgtta tatcctgatt cacgtaatat ttttggggta aatttagttc ctgttccatt
4681 aactgcgcta aaaataattt ttaaatcttt tttagcttct tgctcttttt tgtacgtctc
4741 tgttttagag ctagaaatag caagttaaaa taaggctagt ccgttatcaa cttgaaaaag
4801 tggcaccgag tcggtgcttt tttgaattcg ctagctaggt cttgaaagga gtgggaattg
4861 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
4921 gaggggtcgg caattgatcc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
4981 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
5041 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gaccggttct
5101 agagcgctgc caccatggac aagaagtaca gcatcggcct ggacatcggc accaactctg
5161 tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc aaggtgctgg
5221 gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg ttcgacagcg
5281 gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac accagacgga
5341 agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag gtggacgaca
5401 gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag cacgagcggc
5461 accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac cccaccatct
5521 accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg ctgatctatc
5581 tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc gacctgaacc
5641 ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac aaccagctgt
5701 tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg tctgccagac
5761 tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag aagaagaatg
5821 gcctgttcgg aaacctgatt gccctgagcc tgggcctgac ccccaacttc aagagcaact
5881 tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac gacgacctgg
5941 acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc gccaagaacc
6001 tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc accaaggccc
6061 ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg accctgctga
6121 aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc gaccagagca
6181 agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc tacaagttca
6241 tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag ctgaacagag
6301 aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac cagatccacc
6361 tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc ctgaaggaca
6421 accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg ggccctctgg
6481 ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc atcaccccct
6541 ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc gagcggatga
6601 ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc ctgctgtacg
6661 agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag ggaatgagaa
6721 agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg ttcaagacca
6781 accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc gagtgcttcg
6841 actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc acataccacg
6901 atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac gaggacattc
6961 tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc gaggaacggc
7021 tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag cggcggagat
7081 acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac aagcagtccg
7141 gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac ttcatgcagc
7201 tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag gtgtccggcc
7261 agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc attaagaagg
7321 gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc cggcacaagc
7381 ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag ggacagaaga
7441 acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc agccagatcc
7501 tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac ctgtactacc
7561 tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg ctgtccgact
7621 acgatgtgga ccatatcgtg cctcagagct ttctgaagga cgactccatc gacaacaagg
7681 tgctgaccag aagcgacaag aaccggggca agagcgacaa cgtgccctcc gaagaggtcg
7741 tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt acccagagaa
7801 agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat aaggccggct
7861 tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca cagatcctgg
7921 actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa gtgaaagtga
7981 tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt tacaaagtgc
8041 gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc gtgggaaccg
8101 ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac tacaaggtgt
8161 acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct accgccaagt
8221 acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg gccaacggcg
8281 agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc gtgtgggata
8341 agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg aatatcgtga
8401 aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc aagaggaaca
8461 gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc ggcttcgaca
8521 gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc aagtccaaga
8581 aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc agcttcgaga
8641 agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag gacctgatca
8701 tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga atgctggcct
8761 ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat gtgaacttcc
8821 tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat gagcagaaac
8881 agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag atcagcgagt
8941 tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc gcctacaaca
9001 agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg tttaccctga
9061 ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac cggaagaggt
9121 acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc accggcctgt
9181 acgagacacg gatcgacctg tctcagctgg gaggcgacaa gcgacctgcc gccacaaaga
9241 aggctggaca ggctaagaag aagaaagatt acaaagacga tgacgataag ggatccggcg
9301 caacaaactt ctctctgctg aaacaagccg gagatgtcga agagaatcct ggaccgaccg
9361 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc
9421 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca
9481 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
9541 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
9601 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
9661 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
9721 ggttcctggc caccgtcgga gtctcgcccg accaccaggg caagggtctg ggcagcgccg
9781 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
9841 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
9901 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgaacgcgtt
9961 aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt attcttaact
10021 atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat catgctattg
10081 cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg tctctttatg
10141 aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa
10201 cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact ttcgctttcc
10261 ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc tggacagggg
10321 ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt
10381 ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc tacgtccctt
10441 cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg cggcctcttc
10501 cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc tccccgcgtc
10561 gactttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
10621 ggggggactg gaagggctaa ttcactccca acgaagacaa gatctgcttt ttgcttgtac
10681 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
10741 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
10801 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
10861 cagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
10921 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
10981 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
11041 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
11101 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
11161 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
11221 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
11281 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
11341 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
11401 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
11461 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
11521 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
11581 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
11641 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
11701 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
11761 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
11821 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
11881 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
11941 cccgggagct tgtatatcca ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg
12001 gcatagtata tcggcatagt ataatacgac aaggtgagga actaaaccat ggccaagttg
12061 accagtgccg ttccggtgct caccgcgcgc gacgtcgccg gagcggtcga gttctggacc
12121 gaccggctcg ggttctcccg ggacttcgtg gaggacgact tcgccggtgt ggtccgggac
12181 gacgtgaccc tgttcatcag cgcggtccag gaccaggtgg tgccggacaa caccctggcc
12241 tgggtgtggg tgcgcggcct ggacgagctg tacgccgagt ggtcggaggt cgtgtccacg
12301 aacttccggg acgcctccgg gccggccatg accgagatcg gcgagcagcc gtgggggcgg
12361 gagttcgccc tgcgcgaccc ggccggcaac tgcgtgcact tcgtggccga ggagcaggac
12421 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
12481 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
12541 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
12601 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
12661 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
12721 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
12781 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
12841 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
12901 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
12961 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
13021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
13081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
13141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
13201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
13261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
13321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
13381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
13441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
13501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
13561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
13621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
13681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
13741 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
13801 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
13861 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
13921 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
13981 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
14041 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
14101 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
14161 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
14221 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
14281 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
14341 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
14401 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
14461 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
14521 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
14581 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
14641 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
14701 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
14761 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
14821 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgac
//
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