| 質粒名稱: | dCas9-VPR_P2A_mCherry |
|---|---|
| 出品公司: | Addgene |
| 目錄編號: | 154193 |
| 存儲實驗室: | Anna Obenauf |
|
載體骨架基本信息 |
|
| 載體名稱: | pRRL |
| 載體出品公司: | Clontech |
| 空載體大小: | 4507bp |
| 完整質粒大小: | 14796bp |
| 載體類型: | 哺乳細胞表達, Lentiviral |
| 載體抗性: | 氨芐霉素 |
| 生長溫度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷貝數: | 未知 |
|
插入基因信息 |
|
| 插入基因名稱: | sp-dCas9-VPR_P2A_mCherry |
| 物種來源: | Synthetic |
| 插入大小: | 6579bp |
| 基因突變: | D10A, H840A |
| 啟動子: | EF1as |
| 克隆方法: | Gibson Cloning |
|
文獻引用方法 |
|
| 材料和方法部分: | dCas9-VPR_P2A_mCherry was a gift from Anna Obenauf (Addgene plasmid # 154193 ; //n2t.net/addgene:154193 ; RRID:Addgene_154193) |
| 參考文獻部分: | Isolating live cell clones from barcoded populations using CRISPRa-inducible reporters. Umkehrer C, Holstein F, Formenti L, Jude J, Froussios K, Neumann T, Cronin SM, Haas L, Lipp JJ, Burkard TR, Fellner M, Wiesner T, Zuber J, Obenauf AC. Nat Biotechnol. 2020 Jul 27. pii: 10.1038/s41587-020-0614-0. doi: 10.1038/s41587-020-0614-0. 10.1038/s41587-020-0614-0 PubMed 32719478 |
買家導航
dCas9-VPR_P2A_mCherry載體質粒基本信息
dCas9-VPR_P2A_mCherry質粒圖譜載體圖譜和dCas9-VPR_P2A_mCherry載體序列質粒序列多克隆位點信息
dCas9-VPR_P2A_mCherry質粒載體簡介
dCas9-VPR_P2A_mCherry質粒序列載體序列
LOCUS dCas9-VPR_P2A_mCherry 14796 bp DNA circular SYN 14-APR-2025
DEFINITION Lentiviral expression plasmid encoding dCas9-VPR and a P2A linked
mCherry marker.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14796)
AUTHORS Umkehrer C, Holstein F, Formenti L, Jude J, Froussios K, Neumann T,
Cronin SM, Haas L, Lipp JJ, Burkard TR, Fellner M, Wiesner T, Zuber
J, Obenauf AC
TITLE Isolating live cell clones from barcoded populations using
CRISPRa-inducible reporters.
JOURNAL Nat Biotechnol. 2020 Jul 27. pii: 10.1038/s41587-020-0614-0. doi:
10.1038/s41587-020-0614-0.
PUBMED 32719478
REFERENCE 2 (bases 1 to 14796)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
//gt12365.cn
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Biotechnol. 2020 Jul 27. pii: 10.1038/s41587-020-0614-0. doi:
10.1038/s41587-020-0614-0."
COMMENT Sequence Label: dCas9-VPR_P2A_mCherry
FEATURES Location/Qualifiers
source 1..14796
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter complement(335..546)
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
misc_feature 639..756
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(1054..1098)
/codon_start=1
/product="antigenic peptide corresponding to amino acids
655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
et al., 2013)"
/label=gp41 peptide
/note="recognized by the 2H10 single-chain llama nanobody"
/translation="KNEQELLELDKWASL"
misc_feature 1283..1516
/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 2009..2134
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 2181..2361
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(2379..2577)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(2404..2424)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 2579..2958
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 3130..3149
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 3224..3328
/gene="bla"
/label=AmpR promoter
CDS join(3329..3397,3398..4189)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(3547..3566)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 4360..4948
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 4849..4868
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 5102..5119
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 5236..5257
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter join(5272..5277,5278..5295,5296..5302)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 5310..5326
/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)."
primer_bind 5315..5337
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 5334..5350
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 5334..5350
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 5399..5520
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(5410..5429)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 5464..5483
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(5650..6024)
/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"
promoter complement(6043..6090)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(6138..6467)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 6152..6287
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind complement(6206..6225)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 6452..6472
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(6481..6909)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(6591..6612)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 6803..6822
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 6955..7179
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 7168..7185
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
LTR 7211..7391
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind 7896..7912
/label=pBluescriptKS
/note="For pBluescript vector"
primer_bind 7897..7913
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 7916..8504
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 8022..8033
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 8431..8451
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(8529..9239)
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
primer_bind complement(8807..8826)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 9076..9094
/label=mCherry-R
/note="mCherry, reverse primer"
CDS complement(9240..9296)
/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 complement(9312..9881)
/codon_start=1
/product="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
/label=Rta AD
/translation="RDSREGMFLPKPEAGSAISDVFEGREVCQPKRIRPFHPPGSPWAN
RPLPASLAPTPTGPVHEPVGSLTPAPVPQPLDPAPAVTPEASHLLEDPDEETSQAVKAL
REMADTVIPQKEEAAICGQMDLSHPPPRGHLDELTTTLESMTEDLNLDSPLTPELNEIL
DTFLNDECLLHAMHISTGLSIFDTSLF"
CDS complement(9900..10256)
/codon_start=1
/product="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
/label=RelA (p65) AD
/translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS
EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD
EDFSSIADMDFSALL"
CDS complement(10686..10706)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(10731..10880)
/codon_start=1
/product="tetrameric repeat of the minimal activation
domain of herpes simplex virus VP16 (Beerli et al., 1998)"
/label=VP64
/translation="DALDDFDLDMLGSDALDDFDLDMLGSDALDDFDLDMLGSDALDDF
DLDML"
protein_bind complement(10910..10934)
/gene="mutant version of attB"
/label=attB1
/bound_moiety="BP Clonase(TM)"
/note="recombination site for the Gateway(R) BP reaction"
CDS complement(10971..10991)
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS complement(join(11004..12518,12519..12521,12522..12587,
12588..12593,12594..14796,1..281,282..284,285..311))
/codon_start=1
/product="catalytically dead mutant of the Cas9
endonuclease from the Streptococcus pyogenes Type II
CRISPR/Cas system"
/label=Cas9m4
/note="RNA-guided DNA-binding protein that lacks
endonuclease activity due to the mutations D10A, D839A,
H840A, and N863A (Mali et al., 2013)"
/translation="MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKK
NLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEES
FLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIK
FRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL
ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ
IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR
QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLL
RKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARG
NSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEY
FTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFD
SVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER
LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM
QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGR
HKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY
LYYLQNGRDMYVDQELDINRLSDYDVAAIVPQSFLKDDSIDNKVLTRSDKARGKSDNVP
SEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKH
VAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL
NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKT
EITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSK
ESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGI
TIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNE
LALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILAD
ANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL
DATLIHQSITGLYETRIDLSQLGGD"
ORIGIN
1 gactcctcca gcctatggaa gaaagagtca tccaccttag ccatctcatt actaaagatc
61 tcctgcaggt agcagatccg attctttctg cgggtatatc tgcgccgtgc tgttcttttg
121 agccgcgtgg cttcggccgt ctccccggag tcgaacagga gggcgccaat gaggttcttc
181 tttatgctgt ggcgatcggt attgcccaga actttgaatt ttttgctcgg caccttgtac
241 tcgtccgtaa tgacggccca gccgacgctg tttgtgccga tagcgagccc aatggagtac
301 ttcttgtcca tggtggagcg ctctagaacc ggtcctgtgt tctggcggca aacccgttgc
361 gaaaaagaac gttcacggcg actactgcac ttatatacgg ttctccccca ccctcgggaa
421 aaaggcggag ccagtacacg acatcacttt cccagtttac cccgcgccac cttctctagg
481 caccggatca attgccgacc cctcccccca acttctcggg gactgtgggc gatgtgcgct
541 ctgcccactg acgggcaccg gagccaattc ccactccttt caagacctag ctagctaatt
601 aaccaaactg gatctctgct gtccctgtaa taaacccgaa aattttgaat ttttgtaatt
661 tgtttttgta attctttagt ttgtatgtct gttgctatta tgtctactat tctttcccct
721 gcactgtacc ccccaatccc cccttttctt ttaaaattgt ggatgaatac tgccatttgt
781 ctgcagaatt ggcgcacgca gtgccgatcc gttcactaat cgaatggatc tgtctctgtc
841 tctctctcca ccttcttctt ctattccttc gggcctgtcg ggtcccctcg gggttgggag
901 gtgggtctga aacgataatg gtgaatatcc ctgcctaact ctattcacta tagaaagtac
961 agcaaaaact attcttaaac ctaccaagcc tcctactatc attatgaata attttatata
1021 ccacagccaa tttgttatgt taaaccaatt ccacaaactt gcccatttat ctaattccaa
1081 taattcttgt tcattctttt cttgctggtt ttgcgattct tcaattaagg agtgtattaa
1141 gcttgtgtaa ttgttaattt ctctgtccca ctccatccag gtcgtgtgat tccaaatctg
1201 ttccagagat ttattactcc aactagcatt ccaaggcaca gcagtggtgc aaatgagttt
1261 tccagagcaa ccccaaatcc ccaggagctg ttgatccttt aggtatcttt ccacagccag
1321 gattcttgcc tggagctgct tgatgcccca gactgtgagt tgcaacagat gctgttgcgc
1381 ctcaatagcc ctcagcaaat tgttctgctg ctgcactata ccagacaata attgtctggc
1441 ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt cctgctgctc ccaagaaccc
1501 aaggaacaaa gctcctattc ccactgctct tttttctctc tgcaccactc ttctctttgc
1561 cttggtgggt gctactccta atggttcaat ttttactact ttatatttat ataattcact
1621 tctccaattg tccctcatat ctcctcctcc aggtctgaag atcagcggcc gcttgctgtg
1681 cggtggtctt acttttgttt tgctcttcct ctatcttgtc taaagcttcc ttggtgtctt
1741 ttatctctat cctttgatgc acacaataga gggttgctac tgtattatat aatgatctaa
1801 gttcttctga tcctgtctga agggatggtt gtagctgtcc cagtatttgt ctacagcctt
1861 ctgatgtttc taacaggcca ggattaactg cgaatcgttc tagctccctg cttgcccata
1921 ctatatgttt taatttatat tttttctttc cccctggcct taaccgaatt ttttcccatc
1981 gcgatctaat tctcccccgc ttaatactga cgctctcgca cccatctctc tccttctagc
2041 ctccgctagt caaaattttt ggcgtactca ccagtcgccg cccctcgcct cttgccgtgc
2101 gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc tggtttccct ttcgctttca
2161 agtccctgtt cgggcgccac tgctagagat tttccacact gactaaaagg gtctgaggga
2221 tctctagtta ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa
2281 gctttattga ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat
2341 ctggtctaac cagagagacc cagtacaggc aaaacgcgct gcttatatag acctcccacc
2401 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
2461 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
2521 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
2581 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
2641 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
2701 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
2761 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
2821 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
2881 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
2941 tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac atcgcgaagc agcgcaaaac
3001 gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa gccttgcctt gttgtagctt
3061 aaattttgct cgcgcactac tcagcgacct ccaacacaca agcagggagc agatactggc
3121 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatagggga tcgggagatc
3181 tcccgatccg tcgacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
3241 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
3301 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
3361 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
3421 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
3481 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
3541 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
3601 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
3661 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
3721 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
3781 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
3841 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
3901 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
3961 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
4021 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
4081 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
4141 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
4201 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
4261 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
4321 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
4381 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
4441 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
4501 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
4561 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
4621 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
4681 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
4741 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
4801 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
4861 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
4921 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
4981 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
5041 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
5101 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
5161 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
5221 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
5281 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
5341 cagctatgac catgattacg ccaagctcta gctagaggtc gacggtatac agacatgata
5401 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
5461 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
5521 ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc agccggcgtc
5581 ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat cgaaatctcg
5641 tagcacgtgt cagtcctgct cctcggccac gaagtgcacg cagttgccgg ccgggtcgcg
5701 cagggcgaac tcccgccccc acggctgctc gccgatctcg gtcatggccg gcccggaggc
5761 gtcccggaag ttcgtggaca cgacctccga ccactcggcg tacagctcgt ccaggccgcg
5821 cacccacacc caggccaggg tgttgtccgg caccacctgg tcctggaccg cgctgatgaa
5881 cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc tccacgaagt cccgggagaa
5941 cccgagccgg tcggtccaga actcgaccgc tccggcgacg tcgcgcgcgg tgagcaccgg
6001 aacggcactg gtcaacttgg ccatggttta gttcctcacc ttgtcgtatt atactatgcc
6061 gatatactat gccgatgatt aattgtcaac acgtgctgat cagatccgaa aatggatata
6121 caagctcccg ggagcttttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
6181 tggaatagct cagaggcaga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
6241 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
6301 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
6361 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
6421 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagaatt
6481 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
6541 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
6601 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
6661 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
6721 gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc
6781 cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg
6841 caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac
6901 agggcgcgtg gggatacccc ctagagcccc agctggttct ttccgcctca gaagccatag
6961 agcccaccgc atccccagca tgcctgctat tgtcttccca atcctccccc ttgctgtcct
7021 gccccacccc accccccaga atagaatgac acctactcag acaatgcgat gcaatttcct
7081 cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg
7141 aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat cagcgggttt
7201 aaacgggccc tgctagagat tttccacact gactaaaagg gtctgaggga tctctagtta
7261 ccagagtcac acaacagacg ggcacacact acttgaagca ctcaaggcaa gctttattga
7321 ggcttaagca gtgggttccc tagttagcca gagagctccc aggctcagat ctggtctaac
7381 cagagagacc cagtacagtc cggatgcagc tctcgggcca tgtgatgaaa tgctaggcgg
7441 ctgtcaaacc tccactctaa tacttctctc tccgggtcat ccatcccatg caggctcaca
7501 gggtgtaaca agcgggtgtt ctctccttca ttggcttctt ctaccttctc ttgctcaact
7561 ggtactagct tgtagcacca tccaaaggtc agtggatatc tgatccctgg ccctggtgtg
7621 tagttctgcc aatcagggaa gtagccttgt gtgtggtaga tccacagatc aaggatatct
7681 tgtcttcgtt gggagtgaat tagcccttcc agtcccccct tttcttttaa aaagtggcta
7741 agatctacag ctgccttgta agtcattggt cttaaaggta cctgaggtgt gactggaaaa
7801 cccacctcct cctcctcttg tgcttctagc caggcacaat cagcattggt agctgctgta
7861 ttgctacttg tgattgctcc atgtttttct aggtctcgag gtcgacggta tcgatgcggg
7921 gaggcggccc aaagggagat ccgactcgtc tgagggcgaa ggcgaagacg cggaagaggc
7981 cgcagagccg gcagcaggcc gcgggaagga aggtccgctg gattgagggc cgaagggacg
8041 tagcagaagg acgtcccgcg cagaatccag gtggcaacac aggcgagcag ccaaggaaag
8101 gacgatgatt tccccgacaa caccacggaa ttgtcagtgc ccaacagccg agcccctgtc
8161 cagcagcggg caaggcaggc ggcgatgagt tccgccgtgg caatagggag ggggaaagcg
8221 aaagtcccgg aaaggagctg acaggtggtg gcaatgcccc aaccagtggg ggttgcgtca
8281 gcaaacacag tgcacaccac gccacgttgc ctgacaacgg gccacaactc ctcataaaga
8341 gacagcaacc aggatttata caaggaggag aaaatgaaag ccatacggga agcaatagca
8401 tgatacaaag gcattaaagc agcgtatcca catagcgtaa aaggagcaac atagttaaga
8461 ataccagtca atctttcaca aattttgtaa tccagaggtt gattaccgat aagcttgata
8521 tcgaattctc acttgtacag ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt
8581 tcgtactgtt ccacgatggt gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg
8641 acgttgtagg cgccgggcag ctgcacgggc ttcttggcct tgtaggtggt cttgacctca
8701 gcgtcgtagt ggccgccgtc cttcagcttc agcctctgct tgatctcgcc cttcagggcg
8761 ccgtcctcgg ggtacatccg ctcggaggag gcctcccagc ccatggtctt cttctgcatt
8821 acggggccgt cggaggggaa gttggtgccg cgcagcttca ccttgtagat gaactcgccg
8881 tcctgcaggg aggagtcctg ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg
8941 cgctcccatt tgaagccctc ggggaaggac agcttcaagt agtcggggat gtcggcgggg
9001 tgcttcacgt aggccttgga gccgtacatg aactgagggg acaggatgtc ccaggcgaag
9061 ggcagggggc cacccttggt caccttcagc ttggcggtct gggtgccctc gtaggggcgg
9121 ccctcgccct cgccctcgat ctcgaactcg tggccgttca cggagccctc catgtgcacc
9181 ttgaagcgca tgaactcctt gatgatggcc atgttatcct cctcgccctt gctcaccatg
9241 gggccggggt tctcctccac gtcgccggcc tgcttcagca ggctgaagtt ggtggcgccg
9301 ctgccggatc caaacagaga tgtgtcgaag atggacagtc ctgtgctgat atgcatggca
9361 tgcaagaggc actcgtcgtt caggaaggta tccagaatct cgttcaattc cggggtcagg
9421 ggtgagtcca ggttcagatc ctcggtcatg gactcaagtg tggttgtcag ctcatccaga
9481 tggccccttg ggggcggatg ggaaaggtcc atttggccac agattgcagc ctcttccttc
9541 tggggaatca cagtatcggc catctcccga agggctttga cagcctggct cgtctcttca
9601 tcgggatcct ccaacaggtg actggcctcg ggagtcactg cgggcgctgg atccagtggc
9661 tgagggactg gtgccggggt cagtgacccg actggctcat gtactggacc ggttggtgtt
9721 ggtgcgaggc tggcggggag tgggcggttg gcccatggac ttcctggagg atgaaatggc
9781 cggattcgtt ttggctggca cacctcgcgg ccctcaaaca cgtcactaat agcggagccg
9841 gcctcaggct tcggcaaaaa catcccttcc ctggaatccc ggctgccgct gccagagccc
9901 agcaaggctg agaaatccat atcggcgata gagctgaagt cctcgtcgcc agacagcagt
9961 ccattaggca ggcctggtgc tcccagaggg gcaggagctg gatcaggagg cctctgagcg
10021 cctgtcacga gccgggtgat ggcctcgggg tattccatca gcatgggctc ggtggtgtga
10081 ggggccacag ggatgccctg gttcagcagc tgctggaact cgctgttgtc cacgctggcc
10141 aggtcggtga acacggcagg atcggtgctg tttcccagca gggctcccag atcctcgtcg
10201 tcgaactgca gctgcagcag agcttcagac agtgtgccct cgccggcctg tgtaggttta
10261 ggggctggtg gagccacagc ctgtggaggt ccaggagcca gcacaggcac gggtgctggt
10321 gcctgagcca gtgcagacac catggctgga gctggtgcag gagcaggagc ctgaggcagc
10381 acctgaggag gggctggagc cagagcagag gcctgagaga tctggccgct ggggaacacc
10441 atggtaggga actcgtcgta gttgatggtg ctcaggctgc tggtgaaggg gtaaggctgg
10501 ggggcaggtt ttggcacgct ggcgctggat ctgctgggca cggcgattct tctaggtgga
10561 ggtctagggt cggtggggcc gctgaagggg gacttcttca tgatgctctt gaatgtctcg
10621 taggtccgct tccgcttttc ctcgatccgg tgccggtcgt cggtgtcggg caggtactgg
10681 ctaccaactt tgcgtttctt tttcggagat ccggaacttc tagagttaat cagcatgtcc
10741 aggtcgaaat catcaagggc gtcactgccg agcatgtcga ggtcaaagtc atcaagggca
10801 tccgaaccaa gcatgtcaag gtcaaaatca tcgagggcgt cacttcccag catatccaga
10861 tcaaaatcgt ccaatgcgtc agcccgtccg gaaccgctgg cctctttgta gcctgctttt
10921 ttgtacaaac ttgttgatat caacgcgtca agtcgacgga tccctggcga caccttcctc
10981 ttcttcttgg ggtcagccct gctgtctcca ccgagctgag agaggtcgat tcttgtttca
11041 tagagccccg taattgactg atgaatcagt gtggcgtcca ggacctcctt tgtagaggtg
11101 taccgctttc tgtctatggt ggtgtcgaag tacttgaagg ctgcaggcgc gcccaagttg
11161 gtcagagtaa acaagtggat aatgttttct gcctgctccc tgatgggctt atccctgtgc
11221 ttattgtaag cagaaagcac cttatcgagg ttagcgtcgg cgaggatcac tcttttggag
11281 aattcgctta tttgctcgat gatctcatca aggtagtgtt tgtgttgttc cacgaacagc
11341 tgcttctgct cattatcttc gggagaccct ttgagctttt catagtggct ggccagatac
11401 aagaaattaa cgtatttaga gggcagtgcc agctcgttac ctttctgcag ctcgcccgca
11461 ctagcgagca ttcgtttccg gccgttttca agctcaaaga gagagtactt gggaagctta
11521 atgatgaggt cttttttgac ctctttatat cctttcgcct cgagaaagtc gatggggttt
11581 ttttcgaagc ttgatcgctc catgattgtg atgcccagca gttccttgac gcttttgagt
11641 tttttagact tccctttctc cactttggcc acaaccagta cactgtaagc gactgtagga
11701 gaatcgaatc cgccgtattt cttggggtcc caatcttttt tgcgtgcgat cagcttgtcg
11761 ctgttccttt tcgggaggat actttccttg gagaagcctc cggtctgtac ttcggtcttt
11821 ttaacgatgt tcacctgcgg catggacagg accttccgga ctgtcgcgaa atccctaccc
11881 ttgtcccaca cgatttctcc tgtttctccg tttgtttcga taagtggtcg cttccgaatc
11941 tctccattgg ccagtgtaat ctcggtcttg aaaaaattca taatattgct gtaaaagaag
12001 tacttagcgg tggccttgcc tatttcctgc tcagactttg cgatcatttt cctaacatcg
12061 tacactttat agtctccgta aacaaattca gattcaagct tgggatattt tttgataagt
12121 gcagtgccta ccactgcatt caggtaggca tcatgcgcat ggtggtaatt gttgatctct
12181 ctcaccttat aaaactgaaa gtcctttctg aaatctgaga ccagcttaga cttcagagta
12241 ataactttca cctctcgaat cagtttgtca ttttcatcgt acttggtgtt catgcgtgaa
12301 tcgagaattt gggccacgtg cttggtgatc tggcgtgtct caacaagctg ccttttgatg
12361 aagccggctt tatccaactc agacaggcca cctcgttcag ccttagtcag attatcgaac
12421 ttccgttgtg tgatcagttt ggcgttcagc agctgccgcc aataattttt cattttcttg
12481 acaacttctt ctgaggggac gttatcactc ttccctctag ctttatcgga tcttgtcaac
12541 actttattat caatagaatc atctttgaga aaagactggg gcacgatagc agccacgtcg
12601 tagtcggaga gccgattgat gtccagttcc tgatccacgt acatgtccct gccgttctgc
12661 aggtagtaca ggtagagctt ctcattctga agctgggtgt tttcaactgg gtgttcctta
12721 aggatttggg accccagttc ttttataccc tcttcaatcc tcttcatcct ttccctactg
12781 ttcttctgtc ccttctgggt agtttggttc tctcgggcca tctcgataac gatattctcg
12841 ggcttatgcc ttcccattac tttgacgagt tcatccacga ccttaacggt ctgcagtatt
12901 ccctttttga tagctgggct acctgcaaga ttagcgatgt gctcgtgaag actgtccccc
12961 tggccagaaa cttgtgcttt ctggatgtcc tccttaaagg tgagagagtc atcatggatc
13021 aactgcatga agttccggtt ggcaaatcca tcggacttaa gaaaatccag gattgtcttt
13081 ccactctgct tgtctcggat cccattgatc agttttcttg acagccgccc ccatcctgta
13141 tatcggcgcc tcttgagctg tttcatgact ttgtcgtcga agagatgagc gtaagttttc
13201 aagcgttctt caatcatctc cctatcttca aacaacgtaa gggtgaggac aatgtcctca
13261 agaatgtcct cgttctcctc attgtccagg aagtccttgt ctttaatgat tttcaggaga
13321 tcgtgatacg ttcccaggga tgcgttgaag cgatcctcca ctccgctgat ttcaacagag
13381 tcgaaacatt caatcttttt gaaatagtct tctttgagct gtttcacggt aactttccgg
13441 ttcgtcttga agaggaggtc cacgatagct ttcttctgct ctccagacag gaatgctggc
13501 tttctcatcc cttctgtgac gtatttgacc ttggtgagct cgttataaac tgtgaagtac
13561 tcgtacagca gagagtgttt aggaagcacc ttttcgttag gcagattttt atcaaagtta
13621 gtcatccttt cgatgaagga ctgggcagag gcccccttat ccacgacttc ctcgaagttc
13681 cagggagtga tggtctcttc tgatttgcga gtcatccacg cgaatctgga atttccccgg
13741 gcgagggggc ctacatagta gggtatccga aatgtgagga ttttctcaat cttttccctg
13801 ttatctttca aaaaggggta gaaatcctct tgccgcctga ggatagcgtg cagttcgccc
13861 aggtgaatct ggtgggggat gcttccattg tcgaaagtgc gctgtttgcg caacagatct
13921 tctctgttaa gctttaccag cagctcctcg gtgccgtcca ttttttccaa gatgggctta
13981 ataaatttgt aaaattcctc ctggcttgct ccgccgtcaa tgtatccggc gtagccattt
14041 ttagactgat cgaagaaaat ttccttgtac ttctcaggca gttgctgtct gacaagggcc
14101 ttcagcaaag tcaagtcttg gtggtgctca tcatagcgct tgatcatact agcgctcagc
14161 ggagctttgg tgatctccgt gttcactcgc agaatatcac tcagcagaat ggcgtctgac
14221 aggttctttg ccgccaaaaa aaggtctgcg tactggtcgc cgatctgggc cagcagattg
14281 tcgagatcat catcgtaggt gtctttgctc agttgaagct tggcatcttc ggccaggtcg
14341 aagttagatt taaagttggg ggtcagcccg agtgacaggg cgataagatt accaaacagg
14401 ccgttcttct tctccccagg gagctgtgcg atgaggtttt cgagccgccg ggatttggac
14461 agcctagcgc tcaggattgc tttggcgtca actccggatg cgttgatcgg gttctcttcg
14521 aaaagctgat tgtaagtctg aaccagttgg ataaagagtt tgtcgacatc gctgttgtct
14581 gggttcaggt ccccctcgat gaggaagtgt ccccgaaatt tgatcatatg cgccagcgcg
14641 agatagatca accgcaagtc agccttatca gtactgtcta caagcttctt cctcagatga
14701 tatatggttg ggtacttttc atggtacgcc acctcgtcca cgatattgcc aaagattggg
14761 tggcgctcgt gctttttatc ctcctccacc aaaaag
//
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