| 质粒名称: | lenti-EF1a-dCas9-KRAB-Puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 99372 |
| 存储实验室: | Kristen Brennand |
|
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: | 9192 bp |
| 完整质粒大小: | 14337 bp |
| 载体修饰: | -- |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | -- |
| 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: | dCas9-KRAB-T2A-Puro |
| 物种来源: | -- |
| 插入大小: | 5145 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 吉布森克隆 |
| 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-EF1a-dCas9-KRAB-Puro was a gift from Kristen Brennand (Addgene plasmid # 99372 ; http://n2t.net/addgene:99372 ; RRID:Addgene_99372) |
| 参考文献部分: | Evaluating Synthetic Activation and Repression of Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and Astrocytes. Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I, Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ. Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi: 10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27. 10.1016/j.stemcr.2017.06.012 PubMed 28757163 |
买家导航
lenti-EF1a-dCas9-KRAB-Puro载体质粒基本信息
lenti-EF1a-dCas9-KRAB-Puro质粒图谱载体图谱和lenti-EF1a-dCas9-KRAB-Puro载体序列质粒序列多克隆位点信息
lenti-EF1a-dCas9-KRAB-Puro质粒载体简介
lenti-EF1a-dCas9-KRAB-Puro质粒序列载体序列
LOCUS lenti-EF1a-dCas9-KRAB-Puro 14337 bp ds-DNA circular SYN 07-AUG-2019
DEFINITION 3rd generation lenti vector encoding dCas9-KRAB with 2A puromycin
resistance marker (EF1a-dCas9-KRAB-T2A-Puro-WPRE).
ACCESSION .
VERSION .
KEYWORDS lenti-EF1a-dCas9-KRAB-Puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14337)
AUTHORS Ho SM, Hartley BJ, Flaherty E, Rajarajan P, Abdelaal R, Obiorah I,
Barretto N, Muhammad H, Phatnani HP, Akbarian S, Brennand KJ
TITLE Evaluating Synthetic Activation and Repression of
Neuropsychiatric-Related Genes in hiPSC-Derived NPCs, Neurons, and
Astrocytes.
JOURNAL Stem Cell Reports. 2017 Aug 8;9(2):615-628. doi:
10.1016/j.stemcr.2017.06.012. Epub 2017 Jul 27.
PUBMED 28757163
REFERENCE 2 (bases 1 to 14337)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 7, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..14337
/organism="synthetic DNA construct"
/mol_type="other DNA"
misc_feature 179..412
/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 939..1056
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1186..2364
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
intron 1417..2355
/label=EF-1-alpha intron A
/note="intron upstream of the start codon of human
EF-1-alpha"
primer_bind 2312..2332
/label=EF1a-F
/note="Human elongation factor-1a promoter, forward primer"
CDS 2380..2427
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 2428..6528
/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 6583..6603
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 6628..6822
/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
QLTKPDVILRLEKGEEPWLV"
CDS 6844..6897
/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 6910..7503
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="EYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIERVT
ELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLAAQ
QQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETSAP
RNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind 7400..7420
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 7528..8116
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7581..7601)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(7999..8010)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind complement(8119..8135)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8120..8136)
/label=pBluescriptKS
/note="For pBluescript vector"
LTR 8641..8821
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(8847..8864)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 8853..9077
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 9123..9551
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(9210..9229)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 9420..9441
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(9560..9580)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 9565..9894
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 9745..9880
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 9807..9826
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 9942..9989
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 10008..10382
/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 10512..10633
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(10549..10568)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 10603..10622
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(10682..10698)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10682..10698)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10695..10717)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10706..10722
/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(10730..10760)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10775..10796
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(10913..10930)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11084..11672)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11164..11183)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(11843..12703)
/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 12466..12485
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(12704..12808)
/gene="bla"
/label=AmpR promoter
primer_bind complement(12883..12902)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 13074..13453
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 13455..13653
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 13608..13628
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
LTR 13671..13851
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 13898..14023
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
ORIGIN
1 aagtaagacc accgcacagc aagcggccgc tgatcttcag acctggagga ggagatatga
61 gggacaattg gagaagtgaa ttatataaat ataaagtagt aaaaattgaa ccattaggag
121 tagcacccac caaggcaaag agaagagtgg tgcagagaga aaaaagagca gtgggaatag
181 gagctttgtt ccttgggttc ttgggagcag caggaagcac tatgggcgca gcgtcaatga
241 cgctgacggt acaggccaga caattattgt ctggtatagt gcagcagcag aacaatttgc
301 tgagggctat tgaggcgcaa cagcatctgt tgcaactcac agtctggggc atcaagcagc
361 tccaggcaag aatcctggct gtggaaagat acctaaagga tcaacagctc ctggggattt
421 ggggttgctc tggaaaactc atttgcacca ctgctgtgcc ttggaatgct agttggagta
481 ataaatctct ggaacagatt tggaatcaca cgacctggat ggagtgggac agagaaatta
541 acaattacac aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga
601 atgaacaaga attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa
661 caaattggct gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa
721 gaatagtttt tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat
781 cgtttcagac ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag
841 aaggtggaga gagagacaga gacagatcca ttcgattagt gaacggatcg gcactgcgtg
901 cgccaattct gcagacaaat ggcagtattc atccacaatt ttaaaagaaa aggggggatt
961 ggggggtaca gtgcagggga aagaatagta gacataatag caacagacat acaaactaaa
1021 gaattacaaa aacaaattac aaaaattcaa aattttcggg tttattacag ggacagcaga
1081 gatccagttt ggttaattag ctagctgcaa agatggataa agttttaaac agagaggaat
1141 ctttgcagct aatggacctt ctaggtcttg aaaggagtgg gaattggctc cggtgcccgt
1201 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
1261 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
1321 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
1381 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
1441 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
1501 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
1561 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
1621 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
1681 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
1741 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
1801 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
1861 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
1921 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
1981 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
2041 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
2101 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
2161 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
2221 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
2281 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
2341 tttcttccat ttcaggtgtc gtgacgtacg gccaccatga aaaggccggc ggccacgaaa
2401 aaggccggcc aggcaaaaaa gaaaaaggac aagaagtaca gcatcggcct ggccatcggc
2461 accaactctg tgggctgggc cgtgatcacc gacgagtaca aggtgcccag caagaaattc
2521 aaggtgctgg gcaacaccga ccggcacagc atcaagaaga acctgatcgg agccctgctg
2581 ttcgacagcg gcgaaacagc cgaggccacc cggctgaaga gaaccgccag aagaagatac
2641 accagacgga agaaccggat ctgctatctg caagagatct tcagcaacga gatggccaag
2701 gtggacgaca gcttcttcca cagactggaa gagtccttcc tggtggaaga ggataagaag
2761 cacgagcggc accccatctt cggcaacatc gtggacgagg tggcctacca cgagaagtac
2821 cccaccatct accacctgag aaagaaactg gtggacagca ccgacaaggc cgacctgcgg
2881 ctgatctatc tggccctggc ccacatgatc aagttccggg gccacttcct gatcgagggc
2941 gacctgaacc ccgacaacag cgacgtggac aagctgttca tccagctggt gcagacctac
3001 aaccagctgt tcgaggaaaa ccccatcaac gccagcggcg tggacgccaa ggccatcctg
3061 tctgccagac tgagcaagag cagacggctg gaaaatctga tcgcccagct gcccggcgag
3121 aagaagaatg gcctgttcgg caacctgatt gccctgagcc tgggcctgac ccccaacttc
3181 aagagcaact tcgacctggc cgaggatgcc aaactgcagc tgagcaagga cacctacgac
3241 gacgacctgg acaacctgct ggcccagatc ggcgaccagt acgccgacct gtttctggcc
3301 gccaagaacc tgtccgacgc catcctgctg agcgacatcc tgagagtgaa caccgagatc
3361 accaaggccc ccctgagcgc ctctatgatc aagagatacg acgagcacca ccaggacctg
3421 accctgctga aagctctcgt gcggcagcag ctgcctgaga agtacaaaga gattttcttc
3481 gaccagagca agaacggcta cgccggctac attgacggcg gagccagcca ggaagagttc
3541 tacaagttca tcaagcccat cctggaaaag atggacggca ccgaggaact gctcgtgaag
3601 ctgaacagag aggacctgct gcggaagcag cggaccttcg acaacggcag catcccccac
3661 cagatccacc tgggagagct gcacgccatt ctgcggcggc aggaagattt ttacccattc
3721 ctgaaggaca accgggaaaa gatcgagaag atcctgacct tccgcatccc ctactacgtg
3781 ggccctctgg ccaggggaaa cagcagattc gcctggatga ccagaaagag cgaggaaacc
3841 atcaccccct ggaacttcga ggaagtggtg gacaagggcg cttccgccca gagcttcatc
3901 gagcggatga ccaacttcga taagaacctg cccaacgaga aggtgctgcc caagcacagc
3961 ctgctgtacg agtacttcac cgtgtataac gagctgacca aagtgaaata cgtgaccgag
4021 ggaatgagaa agcccgcctt cctgagcggc gagcagaaaa aggccatcgt ggacctgctg
4081 ttcaagacca accggaaagt gaccgtgaag cagctgaaag aggactactt caagaaaatc
4141 gagtgcttcg actccgtgga aatctccggc gtggaagatc ggttcaacgc ctccctgggc
4201 acataccacg atctgctgaa aattatcaag gacaaggact tcctggacaa tgaggaaaac
4261 gaggacattc tggaagatat cgtgctgacc ctgacactgt ttgaggacag agagatgatc
4321 gaggaacggc tgaaaaccta tgcccacctg ttcgacgaca aagtgatgaa gcagctgaag
4381 cggcggagat acaccggctg gggcaggctg agccggaagc tgatcaacgg catccgggac
4441 aagcagtccg gcaagacaat cctggatttc ctgaagtccg acggcttcgc caacagaaac
4501 ttcatgcagc tgatccacga cgacagcctg acctttaaag aggacatcca gaaagcccag
4561 gtgtccggcc agggcgatag cctgcacgag cacattgcca atctggccgg cagccccgcc
4621 attaagaagg gcatcctgca gacagtgaag gtggtggacg agctcgtgaa agtgatgggc
4681 cggcacaagc ccgagaacat cgtgatcgaa atggccagag agaaccagac cacccagaag
4741 ggacagaaga acagccgcga gagaatgaag cggatcgaag agggcatcaa agagctgggc
4801 agccagatcc tgaaagaaca ccccgtggaa aacacccagc tgcagaacga gaagctgtac
4861 ctgtactacc tgcagaatgg gcgggatatg tacgtggacc aggaactgga catcaaccgg
4921 ctgtccgact acgatgtgga ccacatcgtg cctcagagct ttctgaagga cgactccatc
4981 gacaacaagg tgctgaccag aagcgacaag gcccggggca agagcgacaa cgtgccctcc
5041 gaagaggtcg tgaagaagat gaagaactac tggcggcagc tgctgaacgc caagctgatt
5101 acccagagaa agttcgacaa tctgaccaag gccgagagag gcggcctgag cgaactggat
5161 aaggccggct tcatcaagag acagctggtg gaaacccggc agatcacaaa gcacgtggca
5221 cagatcctgg actcccggat gaacactaag tacgacgaga atgacaagct gatccgggaa
5281 gtgaaagtga tcaccctgaa gtccaagctg gtgtccgatt tccggaagga tttccagttt
5341 tacaaagtgc gcgagatcaa caactaccac cacgcccacg acgcctacct gaacgccgtc
5401 gtgggaaccg ccctgatcaa aaagtaccct aagctggaaa gcgagttcgt gtacggcgac
5461 tacaaggtgt acgacgtgcg gaagatgatc gccaagagcg agcaggaaat cggcaaggct
5521 accgccaagt acttcttcta cagcaacatc atgaactttt tcaagaccga gattaccctg
5581 gccaacggcg agatccggaa gcggcctctg atcgagacaa acggcgaaac cggggagatc
5641 gtgtgggata agggccggga ttttgccacc gtgcggaaag tgctgagcat gccccaagtg
5701 aatatcgtga aaaagaccga ggtgcagaca ggcggcttca gcaaagagtc tatcctgccc
5761 aagaggaaca gcgataagct gatcgccaga aagaaggact gggaccctaa gaagtacggc
5821 ggcttcgaca gccccaccgt ggcctattct gtgctggtgg tggccaaagt ggaaaagggc
5881 aagtccaaga aactgaagag tgtgaaagag ctgctgggga tcaccatcat ggaaagaagc
5941 agcttcgaga agaatcccat cgactttctg gaagccaagg gctacaaaga agtgaaaaag
6001 gacctgatca tcaagctgcc taagtactcc ctgttcgagc tggaaaacgg ccggaagaga
6061 atgctggcct ctgccggcga actgcagaag ggaaacgaac tggccctgcc ctccaaatat
6121 gtgaacttcc tgtacctggc cagccactat gagaagctga agggctcccc cgaggataat
6181 gagcagaaac agctgtttgt ggaacagcac aagcactacc tggacgagat catcgagcag
6241 atcagcgagt tctccaagag agtgatcctg gccgacgcta atctggacaa agtgctgtcc
6301 gcctacaaca agcaccggga taagcccatc agagagcagg ccgagaatat catccacctg
6361 tttaccctga ccaatctggg agcccctgcc gccttcaagt actttgacac caccatcgac
6421 cggaagaggt acaccagcac caaagaggtg ctggacgcca ccctgatcca ccagagcatc
6481 accggcctgt acgagacacg gatcgacctg tctcagctgg gaggcgacag cgctggagga
6541 ggtggaagcg gaggaggagg aagcggagga ggaggtagcg gacctaagaa aaagaggaag
6601 gtggcggccg ctggatccgg acgggctaga acactggtta cgttcaagga cgtgtttgtg
6661 gactttacac gtgaggagtg gaaattgctg gatactgcgc aacaaattgt gtatcgaaat
6721 gtcatgcttg agaattacaa gaacctcgtc agtctcggat accagttgac gaaaccggat
6781 gtgatcctta ggctcgaaaa gggggaagaa ccttggctgg taattaactg tacaggttca
6841 ggggagggaa gaggttccct gctcacatgc ggagacgtag aagagaatcc cggacccatg
6901 acctctagag agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
6961 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
7021 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
7081 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
7141 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
7201 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
7261 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
7321 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
7381 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
7441 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
7501 tgagaattcg atatcaagct tatcggtaat caacctctgg attacaaaat ttgtgaaaga
7561 ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg
7621 cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc
7681 tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc
7741 actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt
7801 tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt
7861 gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg
7921 aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg
7981 tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg
8041 ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt
8101 tgggccgcct ccccgcatcg ataccgtcga cctcgagacc tagaaaaaca tggagcaatc
8161 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
8221 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
8281 gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct aattcactcc
8341 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
8401 tggcagaact acacaccagg gccagggatc agatatccac tgacctttgg atggtgctac
8461 aagctagtac cagttgagca agagaaggta gaagaagcca atgaaggaga gaacacccgc
8521 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt attagagtgg
8581 aggtttgaca gccgcctagc atttcatcac atggcccgag agctgcatcc ggactgtact
8641 gggtctctct ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca
8701 ctgcttaagc ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg
8761 tgtgactctg gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc
8821 agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt
8881 tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc
8941 ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg
9001 tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga
9061 tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta gggggtatcc
9121 ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
9181 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
9241 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
9301 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
9361 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
9421 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
9481 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
9541 taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
9601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
9661 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
9721 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
9781 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct
9841 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc
9901 ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg
9961 catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg gccaagttga
10021 ccagtgccgt tccggtgctc accgcgcgcg acgtcgccgg agcggtcgag ttctggaccg
10081 accggctcgg gttctcccgg gacttcgtgg aggacgactt cgccggtgtg gtccgggacg
10141 acgtgaccct gttcatcagc gcggtccagg accaggtggt gccggacaac accctggcct
10201 gggtgtgggt gcgcggcctg gacgagctgt acgccgagtg gtcggaggtc gtgtccacga
10261 acttccggga cgcctccggg ccggccatga ccgagatcgg cgagcagccg tgggggcggg
10321 agttcgccct gcgcgacccg gccggcaact gcgtgcactt cgtggccgag gagcaggact
10381 gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
10441 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
10501 tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
10561 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
10621 tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat
10681 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
10741 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
10801 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
10861 tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca
10921 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
10981 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
11041 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
11101 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
11161 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
11221 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
11281 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
11341 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
11401 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
11461 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
11521 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
11581 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
11641 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
11701 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
11761 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
11821 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
11881 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
11941 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
12001 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
12061 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
12121 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
12181 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
12241 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
12301 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
12361 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
12421 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
12481 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
12541 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
12601 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
12661 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
12721 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
12781 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg
12841 acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc
12901 cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc
12961 gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct
13021 tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg
13081 attattgact agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
13141 ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc
13201 ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca
13261 ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta
13321 tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta
13381 tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat
13441 cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga
13501 ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca
13561 aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg
13621 taggcgtgta cggtgggagg tctatataag cagcgcgttt tgcctgtact gggtctctct
13681 ggttagacca gatctgagcc tgggagctct ctggctaact agggaaccca ctgcttaagc
13741 ctcaataaag cttgccttga gtgcttcaag tagtgtgtgc ccgtctgttg tgtgactctg
13801 gtaactagag atccctcaga cccttttagt cagtgtggaa aatctctagc agtggcgccc
13861 gaacagggac ttgaaagcga aagggaaacc agaggagctc tctcgacgca ggactcggct
13921 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
13981 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
14041 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
14101 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
14161 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
14221 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
14281 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaa
//
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