| 质粒名称: | pMSCV-LTR-dCas9-VP64-BFP |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 46912 |
| 存储实验室: | Stanley Qi, Jonathan Weissman |
|
载体骨架基本信息 |
|
| 载体名称: | MSCV-puro |
| 载体出品公司: | Clontech |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 6300 bp |
| 完整质粒大小: | 11369 bp |
| 载体类型: | 哺乳动物表达，逆转录病毒，CRISPR |
| 筛选标记: | 嘌呤霉素 |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | dCas9-VP64-BFP fusion |
| 物种来源: | H. sapiens (human) |
| 插入大小: | 5055 bp |
| 启动子 | LTR |
| 标签/融合蛋白: |
3xNLS（插入件上的C端子）
BFP（插入件上的C端子）
VP64域（插入时的C终端）
|
| 克隆方法: | 非连接克隆 |
| 5' 测序引物 | CCCTTGAACCTCCTCGTTCGACC |
| 3' 测序引物 | GAGACGTGCTACTTCCATTTGTC |
|
文献引用方法 |
|
| 材料和方法部分: | pMSCV-LTR-dCas9-VP64-BFP was a gift from Stanley Qi & Jonathan Weissman (Addgene plasmid # 46912 ; http://n2t.net/addgene:46912 ; RRID:Addgene_46912) |
| 参考文献部分: | CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in Eukaryotes. Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE, Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA, Weissman JS, Qi LS. Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi: 10.1016/j.cell.2013.06.044. 10.1016/j.cell.2013.06.044 PubMed 23849981 |
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 pMSCV-LTR-dCas9-VP64-BFP 11369 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Human expression vector containing MSCV LTR promoter, dCas9 that is
fused to 2x NLS, VP64 and tagBFP.
ACCESSION .
VERSION .
KEYWORDS pMSCV-LTR-dCas9-VP64-BFP
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11369)
AUTHORS Gilbert LA, Larson MH, Morsut L, Liu Z, Brar GA, Torres SE,
Stern-Ginossar N, Brandman O, Whitehead EH, Doudna JA, Lim WA,
Weissman JS, Qi LS
TITLE CRISPR-Mediated Modular RNA-Guided Regulation of Transcription in
Eukaryotes.
JOURNAL Cell. 2013 Jul 9. pii: S0092-8674(13)00826-X. doi:
10.1016/j.cell.2013.06.044.
PUBMED 23849981
REFERENCE 2 (bases 1 to 11369)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11369
/organism="synthetic DNA construct"
/mol_type="other DNA"
LTR 1..517
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 581..922
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 989..1405
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
primer_bind 1335..1357
/label=pLXSN 5'
/note="Murine stem cell virus, forward primer. Also called
MSCV"
primer_bind 1373..1389
/label=pBABE 5'
/note="Psi packaging signal, forward primer for pBABE
vectors"
regulatory 1419..1428
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 1425..5528
/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 5538..5558
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5574..5594
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5601..5750
/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"
CDS 5757..5777
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5784..6479
/codon_start=1
/product="monomeric blue fluorescent protein"
/label=TagBFP
/note="mammalian codon-optimized"
/translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
LPSKLGHKLN"
promoter 6514..7013
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
primer_bind complement(6549..6572)
/label=MSCV-rev
/note="Murine stem cell virus, reverse primer"
primer_bind 6946..6965
/label=mPGK-F
/note="Mouse PGK promoter, forward primer"
CDS 7034..7633
/codon_start=1
/gene="pac from Streptomyces alboniger"
/product="puromycin N-acetyltransferase"
/label=PuroR
/note="confers resistance to puromycin"
/translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
primer_bind complement(7034..7053)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 7530..7550
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
LTR 7733..8247
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
primer_bind complement(8416..8432)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8416..8432)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8429..8451)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8440..8456
/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(8464..8494)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8509..8530
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(8647..8664)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(8818..9406)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8898..8917)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(9577..10437)
/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 10200..10219
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(10438..10542)
/gene="bla"
/label=AmpR promoter
primer_bind 10610..10628
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(10666..10688)
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
primer_bind 10788..10807
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 11001..11023
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 11151..11170
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
ORIGIN
1 aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat tttgcaaggc
61 atggaaaata cataactgag aatagagaag ttcagatcaa ggttaggaac agagagacag
121 cagaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
181 gaacagatgg tccccagatg cggtcccgcc ctcagcagtt tctagagaac catcagatgt
241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
361 ccctcactcg gcgcgccagt cctccgatag actgcgtcgc ccgggtaccc gtattcccaa
421 taaagcctct tgctgtttgc atccgaatcg tggactcgct gatccttggg agggtctcct
481 cagattgatt gactgcccac ctcgggggtc tttcatttgg aggttccacc gagatttgga
541 gacccctgcc cagggaccac cgaccccccc gccgggaggt aagctggcca gcggtcgttt
601 cgtgtctgtc tctgtctttg tgcgtgtttg tgccggcatc taatgtttgc gcctgcgtct
661 gtactagtta gctaactagc tctgtatctg gcggacccgt ggtggaactg acgagttctg
721 aacacccggc cgcaaccctg ggagacgtcc cagggacttt gggggccgtt tttgtggccc
781 gacctgagga agggagtcga tgtggaatcc gaccccgtca ggatatgtgg ttctggtagg
841 agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg tttggaaccg
901 aagccgcgcg tcttgtctgc tgcagcgctg cagcatcgtt ctgtgttgtc tctgtctgac
961 tgtgtttctg tatttgtctg aaaattaggg ccagactgtt accactccct taagtttgac
1021 cttaggtcac tggaaagatg tcgagcggat cgctcacaac cagtcggtag atgtcaagaa
1081 gagacgttgg gttaccttct gctctgcaga atggccaacc tttaacgtcg gatggccgcg
1141 agacggcacc tttaaccgag acctcatcac ccaggttaag atcaaggtct tttcacctgg
1201 cccgcatgga cacccagacc aggtccccta catcgtgacc tgggaagcct tggcttttga
1261 cccccctccc tgggtcaagc cctttgtaca ccctaagcct ccgcctcctc ttcctccatc
1321 cgccccgtct ctcccccttg aacctcctcg ttcgaccccg cctcgatcct ccctttatcc
1381 agccctcact ccttctctag gcgccggaat tagatctcgc caccatggac aagaagtatt
1441 ctatcggact ggccatcggg actaatagcg tcgggtgggc cgtgatcact gacgagtaca
1501 aggtgccctc taagaagttc aaggtgctcg ggaacaccga ccggcattcc atcaagaaaa
1561 atctgatcgg agctctcctc tttgattcag gggagaccgc tgaagcaacc cgcctcaagc
1621 ggactgctag acggcggtac accaggagga agaaccggat ttgttacctt caagagatat
1681 tctccaacga aatggcaaag gtcgacgaca gcttcttcca taggctggaa gaatcattcc
1741 tcgtggaaga ggataagaag catgaacggc atcccatctt cggtaatatc gtcgacgagg
1801 tggcctatca cgagaaatac ccaaccatct accatcttcg caaaaagctg gtggactcaa
1861 ccgacaaggc agacctccgg cttatctacc tggccctggc ccacatgatc aagttcagag
1921 gccacttcct gatcgagggc gacctcaatc ctgacaatag cgatgtggat aaactgttca
1981 tccagctggt gcagacttac aaccagctct ttgaagagaa ccccatcaat gcaagcggag
2041 tcgatgccaa ggccattctg tcagcccggc tgtcaaagag ccgcagactt gagaatctta
2101 tcgctcagct gccgggtgaa aagaaaaatg gactgttcgg gaacctgatt gctctttcac
2161 ttgggctgac tcccaatttc aagtctaatt tcgacctggc agaggatgcc aagctgcaac
2221 tgtccaagga cacctatgat gacgatctcg acaacctcct ggcccagatc ggtgaccaat
2281 acgccgacct tttccttgct gctaagaatc tttctgacgc catcctgctg tctgacattc
2341 tccgcgtgaa cactgaaatc accaaggccc ctctttcagc ttcaatgatt aagcggtatg
2401 atgagcacca ccaggacctg accctgctta aggcactcgt ccggcagcag cttccggaga
2461 agtacaagga aatcttcttt gaccagtcaa agaatggata cgccggctac atcgacggag
2521 gtgcctccca agaggaattt tataagttta tcaaacctat ccttgagaag atggacggca
2581 ccgaagagct cctcgtgaaa ctgaatcggg aggatctgct gcggaagcag cgcactttcg
2641 acaatgggag cattccccac cagatccatc ttggggagct tcacgccatc cttcggcgcc
2701 aagaggactt ctaccccttt cttaaggaca acagggagaa gattgagaaa attctcactt
2761 tccgcatccc ctactacgtg ggacccctcg ccagaggaaa tagccggttt gcttggatga
2821 ccagaaagtc agaagaaact atcactccct ggaacttcga agaggtggtg gacaagggag
2881 ccagcgctca gtcattcatc gaacggatga ctaacttcga taagaacctc cccaatgaga
2941 aggtcctgcc gaaacattcc ctgctctacg agtactttac cgtgtacaac gagctgacca
3001 aggtgaaata tgtcaccgaa gggatgagga agcccgcatt cctgtcaggc gaacaaaaga
3061 aggcaattgt ggaccttctg ttcaagacca atagaaaggt gaccgtgaag cagctgaagg
3121 aggactattt caagaaaatt gaatgcttcg actctgtgga gattagcggg gtcgaagatc
3181 ggttcaacgc aagcctgggt acctaccatg atctgcttaa gatcatcaag gacaaggatt
3241 ttctggacaa tgaggagaac gaggacatcc ttgaggacat tgtcctgact ctcactctgt
3301 tcgaggaccg ggaaatgatc gaggagaggc ttaagaccta cgcccatctg ttcgacgata
3361 aagtgatgaa gcaacttaaa cggagaagat ataccggatg gggacgcctt agccgcaaac
3421 tcatcaacgg aatccgggac aaacagagcg gaaagaccat tcttgatttc cttaagagcg
3481 acggattcgc taatcgcaac ttcatgcaac ttatccatga tgattccctg acctttaagg
3541 aggacatcca gaaggcccaa gtgtctggac aaggtgactc actgcacgag catatcgcaa
3601 atctggctgg ttcacccgct attaagaagg gtattctcca gaccgtgaaa gtcgtggacg
3661 agctggtcaa ggtgatgggt cgccataaac cagagaacat tgtcatcgag atggccaggg
3721 aaaaccagac tacccagaag ggacagaaga acagcaggga gcggatgaaa agaattgagg
3781 aagggattaa ggagctcggg tcacagatcc ttaaagagca cccggtggaa aacacccagc
3841 ttcagaatga gaagctctat ctgtactacc ttcaaaatgg acgcgatatg tatgtggacc
3901 aagagcttga tatcaacagg ctctcagact acgacgtgga cgccatcgtc cctcagagct
3961 tcctcaaaga cgactcaatt gacaataagg tgctgactcg ctcagacaag aaccggggaa
4021 agtcagataa cgtgccctca gaggaagtcg tgaaaaagat gaagaactat tggcgccagc
4081 ttctgaacgc aaagctgatc actcagcgga agttcgacaa tctcactaag gctgagaggg
4141 gcggactgag cgaactggac aaagcaggat tcattaaacg gcaacttgtg gagactcggc
4201 agattactaa acatgtcgcc caaatccttg actcacgcat gaataccaag tacgacgaaa
4261 acgacaaact tatccgcgag gtgaaggtga ttaccctgaa gtccaagctg gtcagcgatt
4321 tcagaaagga ctttcaattc tacaaagtgc gggagatcaa taactatcat catgctcatg
4381 acgcatatct gaatgccgtg gtgggaaccg ccctgatcaa gaagtaccca aagctggaaa
4441 gcgagttcgt gtacggagac tacaaggtct acgacgtgcg caagatgatt gccaaatctg
4501 agcaggagat cggaaaggcc accgcaaagt acttcttcta cagcaacatc atgaatttct
4561 tcaagaccga aatcaccctt gcaaacggtg agatccggaa gaggccgctc atcgagacta
4621 atggggagac tggcgaaatc gtgtgggaca agggcagaga tttcgctacc gtgcgcaaag
4681 tgctttctat gcctcaagtg aacatcgtga agaaaaccga ggtgcaaacc ggaggctttt
4741 ctaaggaatc aatcctcccc aagcgcaact ccgacaagct cattgcaagg aagaaggatt
4801 gggaccctaa gaagtacggc ggattcgatt caccaactgt ggcttattct gtcctggtcg
4861 tggctaaggt ggaaaaagga aagtctaaga agctcaagag cgtgaaggaa ctgctgggta
4921 tcaccattat ggagcgcagc tccttcgaga agaacccaat tgactttctc gaagccaaag
4981 gttacaagga agtcaagaag gaccttatca tcaagctccc aaagtatagc ctgttcgaac
5041 tggagaatgg gcggaagcgg atgctcgcct ccgctggcga acttcagaag ggtaatgagc
5101 tggctctccc ctccaagtac gtgaatttcc tctaccttgc aagccattac gagaagctga
5161 aggggagccc cgaggacaac gagcaaaagc aactgtttgt ggagcagcat aagcattatc
5221 tggacgagat cattgagcag atttccgagt tttctaaacg cgtcattctc gctgatgcca
5281 acctcgataa agtccttagc gcatacaata agcacagaga caaaccaatt cgggagcagg
5341 ctgagaatat catccacctg ttcaccctca ccaatcttgg tgcccctgcc gcattcaagt
5401 acttcgacac caccatcgac cggaaacgct atacctccac caaagaagtg ctggacgcca
5461 ccctcatcca ccagagcatc accggacttt acgaaactcg gattgacctc tcacagctcg
5521 gaggggatga gggagctccc aagaaaaagc gcaaggtagg tagttccgga tctccgaaaa
5581 agaaacgcaa agttggtagt gatgctttag acgattttga cttagatatg cttggttcag
5641 acgcgttaga cgacttcgac ctagacatgt taggctcaga tgcattggac gacttcgatt
5701 tagatatgtt gggctccgat gccctagatg actttgatct agatatgcta ggtagtccca
5761 aaaagaagag gaaagtggga tctagcgagc tgattaagga gaacatgcac atgaagctgt
5821 acatggaggg caccgtggac aaccatcact tcaagtgcac atccgagggc gaaggcaagc
5881 cctacgaggg cacccagacc atgagaatca aggtggtcga gggcggccct ctccccttcg
5941 ccttcgacat cctggctact agcttcctct acggcagcaa gaccttcatc aaccacaccc
6001 agggcatccc cgacttcttc aagcagtcct tccctgaggg cttcacatgg gagagagtca
6061 ccacatacga agacgggggc gtgctgaccg ctacccagga caccagcctc caggacggct
6121 gcctcatcta caacgtcaag atcagagggg tgaacttcac atccaacggc cctgtgatgc
6181 agaagaaaac actcggctgg gaggccttca ccgagacgct gtaccccgct gacggcggcc
6241 tggaaggcag aaacgacatg gccctgaagc tcgtgggcgg gagccatctg atcgcaaaca
6301 tcaagaccac atatagatcc aagaaacccg ctaagaacct caagatgcct ggcgtctact
6361 atgtggacta cagactggaa agaatcaagg aggccaacaa cgagacctac gtcgagcagc
6421 acgaggtggc agtggccaga tactgcgacc tccctagcaa actggggcac aagcttaatg
6481 gatcctagac tctcgagagt ctcgaattct accgggtagg ggaggcgctt ttcccaaggc
6541 agtctggagc atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct
6601 ggcctcgcac acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc
6661 ccttcgcgcc accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg
6721 cgtcgtgcag gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca
6781 gcaccgctga gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct
6841 ccttcgcttt ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg
6901 ggctcagggg cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc
6961 aaaagcgcac gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgacctgca
7021 gcccaagctt accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt
7081 ccccagggcc gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac
7141 cgtcgatccg gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg
7201 cgtcgggctc gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg
7261 gaccacgccg gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc
7321 cgagttgagc ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca
7381 ccggcccaag gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg
7441 caagggtctg ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt
7501 gcccgccttc ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt
7561 caccgtcacc gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa
7621 gcccggtgcc tgacgcccgc cccacgaccc gcagcgcccg accgaaagga gcgcacgacc
7681 ccatgcatcg ataaaataaa agattttatt tagtctccag aaaaaggggg gaatgaaaga
7741 ccccacctgt aggtttggca agctagctta agtaacgcca ttttgcaagg catggaaaat
7801 acataactga gaatagagaa gttcagatca aggttaggaa cagagagaca gcagaatatg
7861 ggccaaacag gatatctgtg gtaagcagtt cctgccccgg ctcagggcca agaacagatg
7921 gtccccagat gcggtcccgc cctcagcagt ttctagagaa ccatcagatg tttccagggt
7981 gccccaagga cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc
8041 gcttctgttc gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc
8101 ggcgcgccag tcctccgata gactgcgtcg cccgggtacc cgtgtatcca ataaaccctc
8161 ttgcagttgc atccgacttg tggtctcgct gttccttggg agggtctcct ctgagtgatt
8221 gactacccgt cagcgggggt ctttcatggg taacagtttc ttgaagttgg agaacaacat
8281 tctgagggta ggagtcgaat attaagtaat cctgactcaa ttagccactg ttttgaatcc
8341 acatactcca atactcctga aatagttcat tatggacagc gcagaagagc tggggagaat
8401 taattcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
8461 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
8521 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
8581 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
8641 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
8701 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
8761 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
8821 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
8881 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
8941 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
9001 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
9061 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
9121 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
9181 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
9241 ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt
9301 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
9361 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
9421 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
9481 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
9541 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
9601 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
9661 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
9721 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
9781 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
9841 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
9901 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
9961 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
10021 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
10081 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
10141 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
10201 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
10261 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
10321 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
10381 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
10441 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
10501 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
10561 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
10621 cacgaggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
10681 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
10741 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
10801 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
10861 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
10921 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
10981 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcgc aaggaatggt
11041 gcatgcaagg agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg
11101 ccgaaacaag cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg
11161 gcgatatagg cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg
11221 gcgtagaggc gattagtcca atttgttaaa gacaggatat cagtggtcca ggctctagtt
11281 ttgactcaac aatatcacca gctgaagcct atagagtacg agccatagat aaaataaaag
11341 attttattta gtctccagaa aaagggggg
//
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