| 质粒名称: | LentiCRISPRv2-mCherry |
|---|---|
| 出品公司: | addgene |
| 目录编号: | 99154 |
| 存储实验室: | Agata Smogorzewska |
|
载体骨架基本信息 |
|
| 载体名称: | LentiCRISPRv2 |
| 载体出品公司: | Feng Zhang |
| 空载体大小: | 10000 bp |
| 完整质粒大小: | 14984 bp |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | mCherry |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | NEB Stable |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称1: | Cas9/S. pyogenes CRISPR-Cas9 |
| 物种来源: | Synthetic |
| 插入大小: | 4200 bp |
| 启动子: | EFS-NS |
| 标签/融合蛋白: | FLAG (C terminal on insert) |
| 克隆方法: | 限制性内切酶 |
| 5’克隆位点: | AgeI, XbaI, AfeI(未破坏) |
| 3’克隆位点: | BamHI(未破坏) |
| 5’测序引物 | hEF1aprom-F |
| 3’测序引物 | WPRE-R |
| 插入基因名称2: | mCherry |
| 物种来源: | Discoma sp. |
| 插入大小: | 708bp |
| 克隆方法: | Ligation Independent Cloning |
| 5’测序引物 | ACC AAA GAG GTG CTG GAC G |
| 3’测序引物 | WPRE-R |
|
文献引用方法 |
|
| 材料和方法部分: | LentiCRISPRv2-mCherry was a gift from Agata Smogorzewska (Addgene plasmid # 99154 ; http://n2t.net/addgene:99154 ; RRID:Addgene_99154) |
买家导航
载体基本信息
载体图谱质粒图谱和多克隆位点信息
载体简介
载体序列
LOCUS LentiCRISPRv2-mCherry 14984 bp ds-DNA circular SYN 19-JAN-2022
DEFINITION Lentiviral vector encoding sgRNA cloning site +
hSpCAS9-P2A-mCherry..
ACCESSION .
VERSION .
KEYWORDS LentiCRISPRv2-mCherry
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14984)
TITLE LentiCRISPRv2-mCherry
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 14984)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Jan 19, 2022 from SnapGene Server 1.1.58
http://www.biofeng.com
FEATURES Location/Qualifiers
source 1..14984
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind complement(2..21)
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind 248..268
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(277..705)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(387..408)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 599..618
/label=F1ori-R
/note="F1 origin, reverse primer"
polyA_signal 751..975
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
primer_bind 964..981
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
LTR 1007..1240
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
misc_feature 1312..1900
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS 1418..1429
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
primer_bind 1827..1847
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(1916..2623)
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/label=mCherry
/note="mammalian codon-optimized"
/translation="VSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEGT
QTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFE
DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
EIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERAE
GRHSTGGMDELYK"
primer_bind complement(2194..2213)
/label=mCherry-F
/note="mCherry, forward primer"
primer_bind 2463..2481
/label=mCherry-R
/note="mCherry, reverse primer"
CDS complement(2624..2680)
/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(2690..2713)
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS complement(2714..2761)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS complement(2762..6865)
/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"
regulatory 6862..6871
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
promoter complement(6890..7101)
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
misc_RNA complement(7163..7238)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(9128..9368)
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
primer_bind complement(9178..9197)
/label=LKO.1 5'
/note="Human U6 promoter, forward primer"
primer_bind complement(9348..9368)
/label=hU6-F
/note="Human U6 promoter, forward primer"
misc_feature 9419..9536
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(9834..9878)
/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 10063..10296
/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 10789..10914
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
LTR 10961..11141
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(11159..11357)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind complement(11184..11204)
/label=CMV-F
/note="Human CMV immediate early promoter, forward primer"
enhancer 11359..11738
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind 11910..11929
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 12004..12108
/gene="bla"
/label=AmpR promoter
CDS 12109..12969
/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(12327..12346)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 13140..13728
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 13629..13648
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 13882..13899
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 14016..14037
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 14052..14082
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 14090..14106
/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 14095..14117
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 14114..14130
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 14114..14130
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
polyA_signal 14179..14300
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(14190..14209)
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind 14244..14263
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
CDS complement(14430..14804)
/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(14823..14870)
/label=EM7 promoter
/note="synthetic bacterial promoter "
promoter complement(14918..263)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 14932..83
/label=SV40 ori
/note="SV40 origin of replication"
ORIGIN
1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
241 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
301 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
361 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
421 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
481 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
541 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
601 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
661 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga taccccctag
721 agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc ccagcatgcc
781 tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc cccagaatag
841 aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa aggacagtgg
901 gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca gatggctggc
961 aactagaagg cacagtcgag gctgatcagc gggtttaaac gggccctgct agagattttc
1021 cacactgact aaaagggtct gagggatctc tagttaccag agtcacacaa cagacgggca
1081 cacactactt gaagcactca aggcaagctt tattgaggct taagcagtgg gttccctagt
1141 tagccagaga gctcccaggc tcagatctgg tctaaccaga gagacccagt acaagcaaaa
1201 agcagatctt gtcttcgttg ggagtgaatt agcccttcca gtcccccctt ttcttttaaa
1261 aagtggctaa gatctacagc tgccttgtaa gtcattggtc ttaaagtcga cgcggggagg
1321 cggcccaaag ggagatccga ctcgtctgag ggcgaaggcg aagacgcgga agaggccgca
1381 gagccggcag caggccgcgg gaaggaaggt ccgctggatt gagggccgaa gggacgtagc
1441 agaaggacgt cccgcgcaga atccaggtgg caacacaggc gagcagccaa ggaaaggacg
1501 atgatttccc cgacaacacc acggaattgt cagtgcccaa cagccgagcc cctgtccagc
1561 agcgggcaag gcaggcggcg atgagttccg ccgtggcaat agggaggggg aaagcgaaag
1621 tcccggaaag gagctgacag gtggtggcaa tgccccaacc agtgggggtt gcgtcagcaa
1681 acacagtgca caccacgcca cgttgcctga caacgggcca caactcctca taaagagaca
1741 gcaaccagga tttatacaag gaggagaaaa tgaaagccat acgggaagca atagcatgat
1801 acaaaggcat taaagcagcg tatccacata gcgtaaaagg agcaacatag ttaagaatac
1861 cagtcaatct ttcacaaatt ttgtaatcca gaggttgatt gtcgacttaa cgcgtttact
1921 tgtacagctc gtccatgccg ccggtggagt ggcggccctc ggcgcgttcg tactgttcca
1981 cgatggtgta gtcctcgttg tgggaggtga tgtccaactt gatgttgacg ttgtaggcgc
2041 cgggcagctg cacgggcttc ttggccttgt aggtggtctt gacctcagcg tcgtagtggc
2101 cgccgtcctt cagcttcagc ctctgcttga tctcgccctt cagggcgccg tcctcggggt
2161 acatccgctc ggaggaggcc tcccagccca tggtcttctt ctgcattacg gggccgtcgg
2221 aggggaagtt ggtgccgcgc agcttcacct tgtagatgaa ctcgccgtcc tgcagggagg
2281 agtcctgggt cacggtcacc acgccgccgt cctcgaagtt catcacgcgc tcccacttga
2341 agccctcggg gaaggacagc ttcaagtagt cggggatgtc ggcggggtgc ttcacgtagg
2401 ccttggagcc gtacatgaac tgaggggaca ggatgtccca ggcgaagggc agggggccac
2461 ccttggtcac cttcagcttg gcggtctggg tgccctcgta ggggcggccc tcgccctcgc
2521 cctcgatctc gaactcgtgg ccgttcacgg agccctccat gtgcaccttg aagcgcatga
2581 actccttgat gatggccatg ttatcctcct cgcccttgct caccggtcca ggattctctt
2641 cgacatctcc ggcttgtttc agcagagaga agtttgttgc gccggatccc ttatcgtcat
2701 cgtctttgta atctttcttc ttcttagcct gtccagcctt ctttgtggcg gcaggtcgct
2761 tgtcgcctcc cagctgagac aggtcgatcc gtgtctcgta caggccggtg atgctctggt
2821 ggatcagggt ggcgtccagc acctctttgg tgctggtgta cctcttccgg tcgatggtgg
2881 tgtcaaagta cttgaaggcg gcaggggctc ccagattggt cagggtaaac aggtggatga
2941 tattctcggc ctgctctctg atgggcttat cccggtgctt gttgtaggcg gacagcactt
3001 tgtccagatt agcgtcggcc aggatcactc tcttggagaa ctcgctgatc tgctcgatga
3061 tctcgtccag gtagtgcttg tgctgttcca caaacagctg tttctgctca ttatcctcgg
3121 gggagccctt cagcttctca tagtggctgg ccaggtacag gaagttcaca tatttggagg
3181 gcagggccag ttcgtttccc ttctgcagtt cgccggcaga ggccagcatt ctcttccggc
3241 cgttttccag ctcgaacagg gagtacttag gcagcttgat gatcaggtcc tttttcactt
3301 ctttgtagcc cttggcttcc agaaagtcga tgggattctt ctcgaagctg cttctttcca
3361 tgatggtgat ccccagcagc tctttcacac tcttcagttt cttggacttg cccttttcca
3421 ctttggccac caccagcaca gaataggcca cggtggggct gtcgaagccg ccgtacttct
3481 tagggtccca gtccttcttt ctggcgatca gcttatcgct gttcctcttg ggcaggatag
3541 actctttgct gaagccgcct gtctgcacct cggtcttttt cacgatattc acttggggca
3601 tgctcagcac tttccgcacg gtggcaaaat cccggccctt atcccacacg atctccccgg
3661 tttcgccgtt tgtctcgatc agaggccgct tccggatctc gccgttggcc agggtaatct
3721 cggtcttgaa aaagttcatg atgttgctgt agaagaagta cttggcggta gccttgccga
3781 tttcctgctc gctcttggcg atcatcttcc gcacgtcgta caccttgtag tcgccgtaca
3841 cgaactcgct ttccagctta gggtactttt tgatcagggc ggttcccacg acggcgttca
3901 ggtaggcgtc gtgggcgtgg tggtagttgt tgatctcgcg cactttgtaa aactggaaat
3961 ccttccggaa atcggacacc agcttggact tcagggtgat cactttcact tcccggatca
4021 gcttgtcatt ctcgtcgtac ttagtgttca tccgggagtc caggatctgt gccacgtgct
4081 ttgtgatctg ccgggtttcc accagctgtc tcttgatgaa gccggcctta tccagttcgc
4141 tcaggccgcc tctctcggcc ttggtcagat tgtcgaactt tctctgggta atcagcttgg
4201 cgttcagcag ctgccgccag tagttcttca tcttcttcac gacctcttcg gagggcacgt
4261 tgtcgctctt gccccggttc ttgtcgcttc tggtcagcac cttgttgtcg atggagtcgt
4321 ccttcagaaa gctctgaggc acgatatggt ccacatcgta gtcggacagc cggttgatgt
4381 ccagttcctg gtccacgtac atatcccgcc cattctgcag gtagtacagg tacagcttct
4441 cgttctgcag ctgggtgttt tccacggggt gttctttcag gatctggctg cccagctctt
4501 tgatgccctc ttcgatccgc ttcattctct cgcggctgtt cttctgtccc ttctgggtgg
4561 tctggttctc tctggccatt tcgatcacga tgttctcggg cttgtgccgg cccatcactt
4621 tcacgagctc gtccaccacc ttcactgtct gcaggatgcc cttcttaatg gcggggctgc
4681 cggccagatt ggcaatgtgc tcgtgcaggc tatcgccctg gccggacacc tgggctttct
4741 ggatgtcctc tttaaaggtc aggctgtcgt cgtggatcag ctgcatgaag tttctgttgg
4801 cgaagccgtc ggacttcagg aaatccagga ttgtcttgcc ggactgcttg tcccggatgc
4861 cgttgatcag cttccggctc agcctgcccc agccggtgta tctccgccgc ttcagctgct
4921 tcatcacttt gtcgtcgaac aggtgggcat aggttttcag ccgttcctcg atcatctctc
4981 tgtcctcaaa cagtgtcagg gtcagcacga tatcttccag aatgtcctcg ttttcctcat
5041 tgtccaggaa gtccttgtcc ttgataattt tcagcagatc gtggtatgtg cccagggagg
5101 cgttgaaccg atcttccacg ccggagattt ccacggagtc gaagcactcg attttcttga
5161 agtagtcctc tttcagctgc ttcacggtca ctttccggtt ggtcttgaac agcaggtcca
5221 cgatggcctt tttctgctcg ccgctcagga aggcgggctt tctcattccc tcggtcacgt
5281 atttcacttt ggtcagctcg ttatacacgg tgaagtactc gtacagcagg ctgtgcttgg
5341 gcagcacctt ctcgttgggc aggttcttat cgaagttggt catccgctcg atgaagctct
5401 gggcggaagc gcccttgtcc accacttcct cgaagttcca gggggtgatg gtttcctcgc
5461 tctttctggt catccaggcg aatctgctgt ttcccctggc cagagggccc acgtagtagg
5521 ggatgcggaa ggtcaggatc ttctcgatct tttcccggtt gtccttcagg aatgggtaaa
5581 aatcttcctg ccgccgcaga atggcgtgca gctctcccag gtggatctgg tgggggatgc
5641 tgccgttgtc gaaggtccgc tgcttccgca gcaggtcctc tctgttcagc ttcacgagca
5701 gttcctcggt gccgtccatc ttttccagga tgggcttgat gaacttgtag aactcttcct
5761 ggctggctcc gccgtcaatg tagccggcgt agccgttctt gctctggtcg aagaaaatct
5821 ctttgtactt ctcaggcagc tgctgccgca cgagagcttt cagcagggtc aggtcctggt
5881 ggtgctcgtc gtatctcttg atcatagagg cgctcagggg ggccttggtg atctcggtgt
5941 tcactctcag gatgtcgctc agcaggatgg cgtcggacag gttcttggcg gccagaaaca
6001 ggtcggcgta ctggtcgccg atctgggcca gcaggttgtc caggtcgtcg tcgtaggtgt
6061 ccttgctcag ctgcagtttg gcatcctcgg ccaggtcgaa gttgctcttg aagttggggg
6121 tcaggcccag gctcagggca atcaggtttc cgaacaggcc attcttcttc tcgccgggca
6181 gctgggcgat cagattttcc agccgtctgc tcttgctcag tctggcagac aggatggcct
6241 tggcgtccac gccgctggcg ttgatggggt tttcctcgaa cagctggttg taggtctgca
6301 ccagctggat gaacagcttg tccacgtcgc tgttgtcggg gttcaggtcg ccctcgatca
6361 ggaagtggcc ccggaacttg atcatgtggg ccagggccag atagatcagc cgcaggtcgg
6421 ccttgtcggt gctgtccacc agtttctttc tcaggtggta gatggtgggg tacttctcgt
6481 ggtaggccac ctcgtccacg atgttgccga agatggggtg ccgctcgtgc ttcttatcct
6541 cttccaccag gaaggactct tccagtctgt ggaagaagct gtcgtccacc ttggccatct
6601 cgttgctgaa gatctcttgc agatagcaga tccggttctt ccgtctggtg tatcttcttc
6661 tggcggttct cttcagccgg gtggcctcgg ctgtttcgcc gctgtcgaac agcagggctc
6721 cgatcaggtt cttcttgatg ctgtgccggt cggtgttgcc cagcaccttg aatttcttgc
6781 tgggcacctt gtactcgtcg gtgatcacgg cccagcccac agagttggtg ccgatgtcca
6841 ggccgatgct gtacttcttg tccatggtgg cagcgctcta gaaccggtcc tgtgttctgg
6901 cggcaaaccc gttgcgaaaa agaacgttca cggcgactac tgcacttata tacggttctc
6961 ccccaccctc gggaaaaagg cggagccagt acacgacatc actttcccag tttaccccgc
7021 gccaccttct ctaggcaccg gatcaattgc cgacccctcc ccccaacttc tcggggactg
7081 tgggcgatgt gcgctctgcc cactgacggg caccggagcc aattcccact cctttcaaga
7141 cctagctagc gaattcaaaa aagcaccgac tcggtgccac tttttcaagt tgataacgga
7201 ctagccttat tttaacttgc tatttctagc tctaaaacag agacgtacaa aaaagagcaa
7261 gaagctaaaa aagatttaaa aattattttt agcgcagtta atggaacagg aactaaattt
7321 accccaaaaa tattacgtga atcaggatat aacgttattg aggttgaaga gcatgcattt
7381 gaagatgaaa catttaaaaa tgttgtaaat ccaaatccag aatttgatcc tgcatgaaaa
7441 ataccgcttg aatatggtat taaacatgat gcagatatta ttattatgaa tgacccagat
7501 gctgacagat ttggaatggc aataaaacat gatggtcatt ttgtaagatt agatggaaat
7561 caaacaggac caattttaat tgattgaaaa ttatcaaatc taaaacgctt aaatagcatt
7621 ccaaaaaatc cggctctata ttcaagtttt gtaacaagtg atttgggtga tagaatcgct
7681 catgaaaaat atggagttaa tattgtaaaa actttaactg gatttaaatg aatgggtaga
7741 gaaattgcta aagaagaaga taacggatta aattttgttt ttgcttatga agaaagttat
7801 ggatatgtaa ttgatgactc agctagagat aaagatggaa tacaagcttc tatattaata
7861 gcagaggctg cttgatttta taaaaaacaa aataaaacat tagtagacta tttagaagat
7921 ttatttaaag aaatgggtgc atattacact ttcactttaa acttgaattt taaaccagaa
7981 gaaaagaaat taaaaattga accattaatg aaatcattga gagcaacacc cttaactcaa
8041 attgctggac ttaaagttgt taatgttgaa gactacatcg atggaatgta taatatgcca
8101 ggacaagact tactaaaatt ttatttagaa gataagtcat gatttgctgt tcgcccaagt
8161 ggaactgaac ctaaactaaa aatttatttt ataggtgttg gtgaatctgt tcaaaacgct
8221 aaagttaaag tagacgaaat tattaaagaa ttaaaattaa aaatgaatat ataggagaaa
8281 aaatgaaact aaacaaatat atagatcaca cattattaaa acaagatgct acgaaagctg
8341 aaattaaaca attatgtgat gaagcaattg aatttgattt tgcaacagtt tgtgttaatt
8401 catattgaac aagctattgt aaagaattat taaaaggcac aaatgtagga ataacaaatg
8461 ttgtaggttt tcctctaggt gcatgcacaa cagctacaaa agcattcgaa gtttctgaag
8521 caattaaaga tggtgcaaca gaaattgata tggtattaaa tattggtgca ttaaaagaca
8581 aaaattatga attagtttta gaagacatga aagctgtaaa aaaagcagct ggatcacatg
8641 ttgttaaatg tattatggaa aattgtttat taacaaaaga agaaatcatg aaagcttgtg
8701 aaatagctgt tgaagctgga ttagaatttg ttaaaacatc aacaggattt tcaaaatcag
8761 gtgcaacatt tgaagatgtt aaactaatga agtcagttgt taaagacaat gctttagtta
8821 aagcagctgg tggagttaga acatttgaag atgctcaaaa aatgattgaa gcaggagctg
8881 accgcttagg aacaagtggt ggagtagcta ttattaaagg tgaagaaaac aacgcgagtt
8941 actaaaacta gcgttttttt attttgctca tttttattaa aagtttgcaa aaaggaacat
9001 aaaaattcta attattgata ctaaagttat taaaaagaag attttggttg attttataaa
9061 ggtcatagaa tataatattt tagcatgtgt attttgtgtg ctcatttaca accgtctccg
9121 gtgtttcgtc ctttccacaa gatatataaa gccaagaaat cgaaatactt tcaagttacg
9181 gtaagcatat gatagtccat tttaaaacat aattttaaaa ctgcaaacta cccaagaaat
9241 tattactttc tacgtcacgt attttgtact aatatctttg tgtttacagt caaattaatt
9301 ctaattatct ctctaacagc cttgtatcgt atatgcaaat atgaaggaat catgggaaat
9361 aggccctcgg taccttaatt aaccaaactg gatctctgct gtccctgtaa taaacccgaa
9421 aattttgaat ttttgtaatt tgtttttgta attctttagt ttgtatgtct gttgctatta
9481 tgtctactat tctttcccct gcactgtacc ccccaatccc cccttttctt ttaaaattgt
9541 ggatgaatac tgccatttgt ctgcagaatt ggcgcacgca gtgccgatcc gttcactaat
9601 cgaatggatc tgtctctgtc tctctctcca ccttcttctt ctattccttc gggcctgtcg
9661 ggtcccctcg gggttgggag gtgggtctga aacgataatg gtgaatatcc ctgcctaact
9721 ctattcacta tagaaagtac agcaaaaact attcttaaac ctaccaagcc tcctactatc
9781 attatgaata attttatata ccacagccaa tttgttatgt taaaccaatt ccacaaactt
9841 gcccatttat ctaattccaa taattcttgt tcattctttt cttgctggtt ttgcgattct
9901 tcaattaagg agtgtattaa gcttgtgtaa ttgttaattt ctctgtccca ctccatccag
9961 gtcgtgtgat tccaaatctg ttccagagat ttattactcc aactagcatt ccaaggcaca
10021 gcagtggtgc aaatgagttt tccagagcaa ccccaaatcc ccaggagctg ttgatccttt
10081 aggtatcttt ccacagccag gattcttgcc tggagctgct tgatgcccca gactgtgagt
10141 tgcaacagat gctgttgcgc ctcaatagcc ctcagcaaat tgttctgctg ctgcactata
10201 ccagacaata attgtctggc ctgtaccgtc agcgtcattg acgctgcgcc catagtgctt
10261 cctgctgctc ccaagaaccc aaggaacaaa gctcctattc ccactgctct tttttctctc
10321 tgcaccactc ttctctttgc cttggtgggt gctactccta atggttcaat ttttactact
10381 ttatatttat ataattcact tctccaattg tccctcatat ctcctcctcc aggtctgaag
10441 atcagcggcc gcttgctgtg cggtggtctt acttttgttt tgctcttcct ctatcttgtc
10501 taaagcttcc ttggtgtctt ttatctctat cctttgatgc acacaataga gggttgctac
10561 tgtattatat aatgatctaa gttcttctga tcctgtctga agggatggtt gtagctgtcc
10621 cagtatttgt ctacagcctt ctgatgtttc taacaggcca ggattaactg cgaatcgttc
10681 tagctccctg cttgcccata ctatatgttt taatttatat tttttctttc cccctggcct
10741 taaccgaatt ttttcccatc gcgatctaat tctcccccgc ttaatactga cgctctcgca
10801 cccatctctc tccttctagc ctccgctagt caaaattttt ggcgtactca ccagtcgccg
10861 cccctcgcct cttgccgtgc gcgcttcagc aagccgagtc ctgcgtcgag agagctcctc
10921 tggtttccct ttcgctttca agtccctgtt cgggcgccac tgctagagat tttccacact
10981 gactaaaagg gtctgaggga tctctagtta ccagagtcac acaacagacg ggcacacact
11041 acttgaagca ctcaaggcaa gctttattga ggcttaagca gtgggttccc tagttagcca
11101 gagagctccc aggctcagat ctggtctaac cagagagacc cagtacaggc aaaacgcgct
11161 gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg gggcggagtt
11221 gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc cattgacgtc
11281 aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc cattgatgta
11341 ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt actgccaagt
11401 aggaaagtcc cataaggtca tgtactgggc ataatgccag gcgggccatt taccgtcatt
11461 gacgtcaata gggggcgtac ttggcatatg atacacttga tgtactgcca agtgggcagt
11521 ttaccgtaaa tactccaccc attgacgtca atggaaagtc cctattggcg ttactatggg
11581 aacatacgtc attattgacg tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc
11641 atttaccgta agttatgtaa cgcggaactc catatatggg ctatgaacta atgaccccgt
11701 aattgattac tattaataac tagtcaataa tcaatgtcaa cgcgtatatc tggcccgtac
11761 atcgcgaagc agcgcaaaac gcctaaccct aagcagattc ttcatgcaat tgtcggtcaa
11821 gccttgcctt gttgtagctt aaattttgct cgcgcactac tcagcgacct ccaacacaca
11881 agcagggagc agatactggc ttaactatgc ggcatcagag cagattgtac tgagagtgca
11941 ccatagggga tcgggagatc tcccgatccg tcgacgtcag gtggcacttt tcggggaaat
12001 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
12061 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
12121 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
12181 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
12241 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
12301 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc
12361 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
12421 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
12481 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
12541 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
12601 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg
12661 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
12721 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
12781 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
12841 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
12901 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
12961 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
13021 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
13081 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
13141 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
13201 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
13261 agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc
13321 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
13381 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
13441 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
13501 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
13561 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
13621 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
13681 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
13741 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
13801 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
13861 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata
13921 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
13981 cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag
14041 gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga
14101 taacaatttc acacaggaaa cagctatgac catgattacg ccaagctcta gctagaggtc
14161 gacggtatac agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc
14221 agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta
14281 taagctgcaa taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg
14341 aggatcatcc agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg
14401 gcggtggaat cgaaatctcg tagcacgtgt cagtcctgct cctcggccac gaagtgcacg
14461 cagttgccgg ccgggtcgcg cagggcgaac tcccgccccc acggctgctc gccgatctcg
14521 gtcatggccg gcccggaggc gtcccggaag ttcgtggaca cgacctccga ccactcggcg
14581 tacagctcgt ccaggccgcg cacccacacc caggccaggg tgttgtccgg caccacctgg
14641 tcctggaccg cgctgatgaa cagggtcacg tcgtcccgga ccacaccggc gaagtcgtcc
14701 tccacgaagt cccgggagaa cccgagccgg tcggtccaga actcgaccgc tccggcgacg
14761 tcgcgcgcgg tgagcaccgg aacggcactg gtcaacttgg ccatggttta gttcctcacc
14821 ttgtcgtatt atactatgcc gatatactat gccgatgatt aattgtcaac acgtgctgat
14881 cagatccgaa aatggatata caagctcccg ggagcttttt gcaaaagcct aggcctccaa
14941 aaaagcctcc tcactacttc tggaatagct cagaggcaga ggcg
//
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