| 质粒名称: | lentiCRISPRv2 hygro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 98291 |
| 存储实验室: | Brett Stringer |
|
载体骨架基本信息 |
|
| 载体名称: | lentiCRISPRv2 |
| 载体出品公司: | Feng Zhang (Addgene #52961) |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 15261 bp |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 慢病毒，CRISPR |
| 筛选标记: | 潮霉素 |
| 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: | P2A-hygro |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | EF-1a |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | BamHI(未破坏) |
| 3' 克隆位点: | MluI(未破坏) |
| 5' 测序引物 | CCAAAGAGGTGCTGGACG |
| 3' 测序引物 | WPRE-R (CATAGCGTAAAAGGAGCAACA) |
|
文献引用方法 |
|
| 材料和方法部分: | lentiCRISPRv2 hygro was a gift from Brett Stringer (Addgene plasmid # 98291 ; http://n2t.net/addgene:98291 ; RRID:Addgene_98291) |
| 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
lentiCRISPRv2hygro载体基本信息
lentiCRISPRv2hygro载体图谱质粒图谱和多克隆位点信息
lentiCRISPRv2hygro载体简介
lentiCRISPRv2hygro载体序列
LOCUS lentiCRISPRv2 hygro 15261 bp ds-DNA circular SYN 09-NOV-2018
DEFINITION Variant of lentiCRISPRv2 that confers hygromycin resistance.
ACCESSION .
VERSION .
KEYWORDS lentiCRISPRv2 hygro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 15261)
AUTHORS Brett Stringer
TITLE Brett Stringer Lab Plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 15261)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Nov 9, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..15261
/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"
misc_feature 10353..10941
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(10406..10426)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(10824..10835)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 11013..11246
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(11272..11289)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 11278..11502
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 11548..11976
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(11635..11654)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 11845..11866
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(11985..12005)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 11990..12319
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 12170..12305
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 12232..12251
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
promoter 12367..12414
/label=EM7 promoter
/note="synthetic bacterial promoter "
CDS 12433..12807
/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 12937..13058
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(12974..12993)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 13028..13047
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(13107..13123)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(13107..13123)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(13120..13142)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 13131..13147
/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(13155..13185)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 13200..13221
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(13338..13355)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(13509..14097)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(13589..13608)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(14268..15128)
/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 14891..14910
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(15129..15233)
/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 ggaccgaaaa
9361 agcctgaact caccgcgacg tctgtcgaga agtttctgat cgaaaagttc gacagcgtgt
9421 ccgacctgat gcagctctcg gagggcgaag aatctcgtgc tttcagcttc gatgtaggag
9481 ggcgtggata tgtcctgcgg gtaaatagct gcgccgatgg tttctacaaa gatcgttatg
9541 tttatcggca ctttgcatcg gccgcgctcc cgattccgga agtgcttgac attggggaat
9601 tcagcgagag cctgacctat tgcatctccc gccgtgcaca gggtgtcacg ttgcaagacc
9661 tgcctgaaac cgaactgccc gctgttctgc agccggtcgc ggaggccatg gatgcgatcg
9721 ctgcggccga tcttagccag acgagcgggt tcggcccatt cggaccgcaa ggaatcggtc
9781 aatacactac atggcgtgat ttcatatgcg cgattgctga tccccatgtg tatcactggc
9841 aaactgtgat ggacgacacc gtcagtgcgt ccgtcgcgca ggctctcgat gagctgatgc
9901 tttgggccga ggactgcccc gaagtccggc acctcgtgca cgcggatttc ggctccaaca
9961 atgtcctgac ggacaatggc cgcataacag cggtcattga ctggagcgag gcgatgttcg
10021 gggattccca atacgaggtc gccaacatct tcttctggag gccgtggttg gcttgtatgg
10081 agcagcagac gcgctacttc gagcggaggc atccggagct tgcaggatcg ccgcggctcc
10141 gggcgtatat gctccgcatt ggtcttgacc aactctatca gagcttggtt gacggcaatt
10201 tcgatgatgc agcttgggcg cagggtcgat gcgacgcaat cgtccgatcc ggagccggga
10261 ctgtcgggcg tacacaaatc gcccgcagaa gcgcggccgt ctggaccgat ggctgtgtag
10321 aagtacttcg ccgatagacg cgttaagtcg acaatcaacc tctggattac aaaatttgtg
10381 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
10441 taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata
10501 aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg
10561 tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc acctgtcagc
10621 tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc atcgccgcct
10681 gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc gtggtgttgt
10741 cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg attctgcgcg
10801 ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct tcccgcggcc
10861 tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg agtcggatct
10921 ccctttgggc cgcctccccg cgtcgacttt aagaccaatg acttacaagg cagctgtaga
10981 tcttagccac tttttaaaag aaaagggggg actggaaggg ctaattcact cccaacgaag
11041 acaagatctg ctttttgctt gtactgggtc tctctggtta gaccagatct gagcctggga
11101 gctctctggc taactaggga acccactgct taagcctcaa taaagcttgc cttgagtgct
11161 tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac tagagatccc tcagaccctt
11221 ttagtcagtg tggaaaatct ctagcagggc ccgtttaaac ccgctgatca gcctcgactg
11281 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
11341 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
11401 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
11461 aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa
11521 ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg
11581 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
11641 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
11701 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
11761 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
11821 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
11881 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
11941 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
12001 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
12061 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
12121 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
12181 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
12241 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
12301 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
12361 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
12421 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
12481 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
12541 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
12601 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
12661 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
12721 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
12781 cacttcgtgg ccgaggagca ggactgacac gtgctacgag atttcgattc caccgccgcc
12841 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
12901 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
12961 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
13021 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
13081 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
13141 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
13201 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
13261 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
13321 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
13381 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
13441 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
13501 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
13561 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
13621 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
13681 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
13741 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
13801 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
13861 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
13921 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
13981 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
14041 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
14101 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
14161 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
14221 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
14281 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
14341 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
14401 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
14461 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
14521 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
14581 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
14641 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
14701 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
14761 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
14821 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
14881 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
14941 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
15001 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
15061 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
15121 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
15181 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
15241 ccccgaaaag tgccacctga c
//
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