| 质粒名称: | LentiV_Cas9_puro |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 108100 |
| 存储实验室: | PUBLICATION |
|
载体骨架基本信息 |
|
| 载体名称: | LentiV_puro |
| 载体出品公司: | -- |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | 7000 bp |
| 完整质粒大小: | 11203bp |
| 载体修饰: | 人细胞色素c氧化酶亚基VIII（human cytochrome C oxidase subunit VIII） |
| 载体类型: | 哺乳细胞表达 |
| 筛选标记: | 新霉素（G418） |
| 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: | Cas9 |
| 物种来源: | 光合细菌 Rhodopseudomonas palustris |
| 插入大小: | 4100bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | AgeI(未破坏) |
| 3' 克隆位点: | NotI (未破坏) |
| 5' 测序引物 | CMV-F |
| 3' 测序引物 | SV40pA-R |
|
文献引用方法 |
|
| 材料和方法部分: | LentiV_Cas9_puro was a gift from Christopher Vakoc (Addgene plasmid # 108100 ; http://n2t.net/addgene:108100 ; RRID:Addgene_108100) |
| 参考文献部分: | LKB1, Salt-Inducible Kinases, and MEF2C Are Linked Dependencies in Acute Myeloid Leukemia. Tarumoto Y, Lu B, Somerville TDD, Huang YH, Milazzo JP, Wu XS, Klingbeil O, El Demerdash O, Shi J, Vakoc CR. Mol Cell. 2018 Feb 28. pii: S1097-2765(18)30109-6. doi: 10.1016/j.molcel.2018.02.011. 10.1016/j.molcel.2018.02.011 PubMed 29526696 |
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 LentiV_Cas9_puro 11203 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Lentiviral expression plasmid of spCas9 with puromycin resistance
gene.
ACCESSION .
VERSION .
KEYWORDS LentiV_Cas9_puro
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11203)
AUTHORS Tarumoto Y, Lu B, Somerville TDD, Huang YH, Milazzo JP, Wu XS,
Klingbeil O, El Demerdash O, Shi J, Vakoc CR
TITLE LKB1, Salt-Inducible Kinases, and MEF2C Are Linked Dependencies in
Acute Myeloid Leukemia.
JOURNAL Mol Cell. 2018 Feb 28. pii: S1097-2765(18)30109-6. doi:
10.1016/j.molcel.2018.02.011.
PUBMED 29526696
REFERENCE 2 (bases 1 to 11203)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..11203
/organism="synthetic DNA construct"
/mol_type="other DNA"
promoter 12..238
/label=RSV promoter
/note="Rous sarcoma virus enhancer/promoter"
LTR 239..419
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 466..591
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1084..1317
/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 1730..1847
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 1916..2127
/label=EF-1-alpha core promoter
/note="core promoter for human elongation factor
EF-1-alpha"
regulatory 2147..2156
/regulatory_class="other"
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
CDS 2156..2179
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 2186..2206
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 2231..6331
/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="DKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKN
LIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF
LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF
RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE
NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI
GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQ
QLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLR
KQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGN
SRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYF
TVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDS
VEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERL
KTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQ
LIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRH
KPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL
YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPS
EEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV
AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLN
AVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTE
ITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE
SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGIT
IMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNEL
ALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADA
NLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLD
ATLIHQSITGLYETRIDLSQLGGD"
CDS 6332..6379
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label=nucleoplasmin NLS
/translation="KRPAATKKAGQAKKKK"
CDS 6395..6451
/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 6452..7051
/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(6452..6471)
/label=Puro-R
/note="Puromycin resistance gene, reverse primer. Also
called puro-variant-R"
primer_bind 6948..6968
/label=Puro-F
/note="Puromycin resistance gene, forward primer"
misc_feature 7067..7655
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(7120..7140)
/label=WPRE-R
/note="WPRE, reverse primer"
CDS complement(7538..7549)
/codon_start=1
/product="Factor Xa recognition and cleavage site"
/label=Factor Xa site
/translation="IEGR"
LTR 7727..7960
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 8038..8159
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(8075..8094)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 8129..8148
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
rep_origin 8199..8334
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 8261..8280
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
primer_bind complement(8354..8373)
/label=T7
/note="T7 promoter, forward primer"
promoter complement(8355..8373)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8383..8400)
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(8383..8399)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8392..8414)
/label=M13/pUC Forward
/note="In lacZ gene"
rep_origin 8541..8996
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(8628..8647)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 8838..8859
/label=F1ori-F
/note="F1 origin, forward primer"
promoter 9022..9126
/gene="bla"
/label=AmpR promoter
CDS 9127..9987
/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(9345..9364)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 10158..10746
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 10647..10666
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 10900..10917
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 11034..11055
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 11070..11100
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 11108..11124
/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 11113..11135
/label=M13/pUC Reverse
/note="In lacZ gene"
primer_bind 11132..11148
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 11132..11148
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind 11167..11187
/label=T3
/note="T3 promoter, forward primer"
promoter 11169..11187
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
1 tgcaagctta atgtagtctt atgcaatact cttgtagtct tgcaacatgg taacgatgag
61 ttagcaacat gccttacaag gagagaaaaa gcaccgtgca tgccgattgg tggaagtaag
121 gtggtacgat cgtgccttat taggaaggca acagacgggt ctgacatgga ttggacgaac
181 cactgaattg ccgcattgca gagatattgt atttaagtgc ctagctcgat acataaacgg
241 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
301 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
361 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
421 tggcgcccga acagggactt gaaagcgaaa gggaaaccag aggagctctc tcgacgcagg
481 actcggcttg ctgaagcgcg cacggcaaga ggcgaggggc ggcgactggt gagtacgcca
541 aaaattttga ctagcggagg ctagaaggag agagatgggt gcgagagcgt cagtattaag
601 cgggggagaa ttagatcgcg atgggaaaaa attcggttaa ggccaggggg aaagaaaaaa
661 tataaattaa aacatatagt atgggcaagc agggagctag aacgattcgc agttaatcct
721 ggcctgttag aaacatcaga aggctgtaga caaatactgg gacagctaca accatccctt
781 cagacaggat cagaagaact tagatcatta tataatacag tagcaaccct ctattgtgtg
841 catcaaagga tagagataaa agacaccaag gaagctttag acaagataga ggaagagcaa
901 aacaaaagta agaccaccgc acagcaagcg gccgctgatc ttcagacctg gaggaggaga
961 tatgagggac aattggagaa gtgaattata taaatataaa gtagtaaaaa ttgaaccatt
1021 aggagtagca cccaccaagg caaagagaag agtggtgcag agagaaaaaa gagcagtggg
1081 aataggagct ttgttccttg ggttcttggg agcagcagga agcactatgg gcgcagcgtc
1141 aatgacgctg acggtacagg ccagacaatt attgtctggt atagtgcagc agcagaacaa
1201 tttgctgagg gctattgagg cgcaacagca tctgttgcaa ctcacagtct ggggcatcaa
1261 gcagctccag gcaagaatcc tggctgtgga aagataccta aaggatcaac agctcctggg
1321 gatttggggt tgctctggaa aactcatttg caccactgct gtgccttgga atgctagttg
1381 gagtaataaa tctctggaac agatttggaa tcacacgacc tggatggagt gggacagaga
1441 aattaacaat tacacaagct taatacactc cttaattgaa gaatcgcaaa accagcaaga
1501 aaagaatgaa caagaattat tggaattaga taaatgggca agtttgtgga attggtttaa
1561 cataacaaat tggctgtggt atataaaatt attcataatg atagtaggag gcttggtagg
1621 tttaagaata gtttttgctg tactttctat agtgaataga gttaggcagg gatattcacc
1681 attatcgttt cagacccacc tcccaacccc gaggggaccc agaccacaat tttaaaagaa
1741 aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata gcaacagaca
1801 tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg gtttattaca
1861 gggacagcag agatccactt tggcgccggc tcgagtggct ccggtgcccg tcagtgggca
1921 gagcgcacat cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt
1981 gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt
2041 tttcccgagg gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt
2101 cgcaacgggt ttgccgccag aacacaggtg tcgtgacgcg ggatccgcca ccatggatta
2161 caaagacgat gacgataaga tggccccaaa gaagaagcgg aaggtcggta tccacggagt
2221 cccagcagcc gacaagaagt acagcatcgg cctggacatc ggcaccaact ctgtgggctg
2281 ggccgtgatc accgacgagt acaaggtgcc cagcaagaaa ttcaaggtgc tgggcaacac
2341 cgaccggcac agcatcaaga agaacctgat cggagccctg ctgttcgaca gcggcgaaac
2401 agccgaggcc acccggctga agagaaccgc cagaagaaga tacaccagac ggaagaaccg
2461 gatctgctat ctgcaagaga tcttcagcaa cgagatggcc aaggtggacg acagcttctt
2521 ccacagactg gaagagtcct tcctggtgga agaggataag aagcacgagc ggcaccccat
2581 cttcggcaac atcgtggacg aggtggccta ccacgagaag taccccacca tctaccacct
2641 gagaaagaaa ctggtggaca gcaccgacaa ggccgacctg cggctgatct atctggccct
2701 ggcccacatg atcaagttcc ggggccactt cctgatcgag ggcgacctga accccgacaa
2761 cagcgacgtg gacaagctgt tcatccagct ggtgcagacc tacaaccagc tgttcgagga
2821 aaaccccatc aacgccagcg gcgtggacgc caaggccatc ctgtctgcca gactgagcaa
2881 gagcagacgg ctggaaaatc tgatcgccca gctgcccggc gagaagaaga atggcctgtt
2941 cggaaacctg attgccctga gcctgggcct gacccccaac ttcaagagca acttcgacct
3001 ggccgaggat gccaaactgc agctgagcaa ggacacctac gacgacgacc tggacaacct
3061 gctggcccag atcggcgacc agtacgccga cctgtttctg gccgccaaga acctgtccga
3121 cgccatcctg ctgagcgaca tcctgagagt gaacaccgag atcaccaagg cccccctgag
3181 cgcctctatg atcaagagat acgacgagca ccaccaggac ctgaccctgc tgaaagctct
3241 cgtgcggcag cagctgcctg agaagtacaa agagattttc ttcgaccaga gcaagaacgg
3301 ctacgccggc tacattgacg gcggagccag ccaggaagag ttctacaagt tcatcaagcc
3361 catcctggaa aagatggacg gcaccgagga actgctcgtg aagctgaaca gagaggacct
3421 gctgcggaag cagcggacct tcgacaacgg cagcatcccc caccagatcc acctgggaga
3481 gctgcacgcc attctgcggc ggcaggaaga tttttaccca ttcctgaagg acaaccggga
3541 aaagatcgag aagatcctga ccttccgcat cccctactac gtgggccctc tggccagggg
3601 aaacagcaga ttcgcctgga tgaccagaaa gagcgaggaa accatcaccc cctggaactt
3661 cgaggaagtg gtggacaagg gcgcttccgc ccagagcttc atcgagcgga tgaccaactt
3721 cgataagaac ctgcccaacg agaaggtgct gcccaagcac agcctgctgt acgagtactt
3781 caccgtgtat aacgagctga ccaaagtgaa atacgtgacc gagggaatga gaaagcccgc
3841 cttcctgagc ggcgagcaga aaaaggccat cgtggacctg ctgttcaaga ccaaccggaa
3901 agtgaccgtg aagcagctga aagaggacta cttcaagaaa atcgagtgct tcgactccgt
3961 ggaaatctcc ggcgtggaag atcggttcaa cgcctccctg ggcacatacc acgatctgct
4021 gaaaattatc aaggacaagg acttcctgga caatgaggaa aacgaggaca ttctggaaga
4081 tatcgtgctg accctgacac tgtttgagga cagagagatg atcgaggaac ggctgaaaac
4141 ctatgcccac ctgttcgacg acaaagtgat gaagcagctg aagcggcgga gatacaccgg
4201 ctggggcagg ctgagccgga agctgatcaa cggcatccgg gacaagcagt ccggcaagac
4261 aatcctggat ttcctgaagt ccgacggctt cgccaacaga aacttcatgc agctgatcca
4321 cgacgacagc ctgaccttta aagaggacat ccagaaagcc caggtgtccg gccagggcga
4381 tagcctgcac gagcacattg ccaatctggc cggcagcccc gccattaaga agggcatcct
4441 gcagacagtg aaggtggtgg acgagctcgt gaaagtgatg ggccggcaca agcccgagaa
4501 catcgtgatc gaaatggcca gagagaacca gaccacccag aagggacaga agaacagccg
4561 cgagagaatg aagcggatcg aagagggcat caaagagctg ggcagccaga tcctgaaaga
4621 acaccccgtg gaaaacaccc agctgcagaa cgagaagctg tacctgtact acctgcagaa
4681 tgggcgggat atgtacgtgg accaggaact ggacatcaac cggctgtccg actacgatgt
4741 ggaccatatc gtgcctcaga gctttctgaa ggacgactcc atcgacaaca aggtgctgac
4801 cagaagcgac aagaaccggg gcaagagcga caacgtgccc tccgaagagg tcgtgaagaa
4861 gatgaagaac tactggcggc agctgctgaa cgccaagctg attacccaga gaaagttcga
4921 caatctgacc aaggccgaga gaggcggcct gagcgaactg gataaggccg gcttcatcaa
4981 gagacagctg gtggaaaccc ggcagatcac aaagcacgtg gcacagatcc tggactcccg
5041 gatgaacact aagtacgacg agaatgacaa gctgatccgg gaagtgaaag tgatcaccct
5101 gaagtccaag ctggtgtccg atttccggaa ggatttccag ttttacaaag tgcgcgagat
5161 caacaactac caccacgccc acgacgccta cctgaacgcc gtcgtgggaa ccgccctgat
5221 caaaaagtac cctaagctgg aaagcgagtt cgtgtacggc gactacaagg tgtacgacgt
5281 gcggaagatg atcgccaaga gcgagcagga aatcggcaag gctaccgcca agtacttctt
5341 ctacagcaac atcatgaact ttttcaagac cgagattacc ctggccaacg gcgagatccg
5401 gaagcggcct ctgatcgaga caaacggcga aaccggggag atcgtgtggg ataagggccg
5461 ggattttgcc accgtgcgga aagtgctgag catgccccaa gtgaatatcg tgaaaaagac
5521 cgaggtgcag acaggcggct tcagcaaaga gtctatcctg cccaagagga acagcgataa
5581 gctgatcgcc agaaagaagg actgggaccc taagaagtac ggcggcttcg acagccccac
5641 cgtggcctat tctgtgctgg tggtggccaa agtggaaaag ggcaagtcca agaaactgaa
5701 gagtgtgaaa gagctgctgg ggatcaccat catggaaaga agcagcttcg agaagaatcc
5761 catcgacttt ctggaagcca agggctacaa agaagtgaaa aaggacctga tcatcaagct
5821 gcctaagtac tccctgttcg agctggaaaa cggccggaag agaatgctgg cctctgccgg
5881 cgaactgcag aagggaaacg aactggccct gccctccaaa tatgtgaact tcctgtacct
5941 ggccagccac tatgagaagc tgaagggctc ccccgaggat aatgagcaga aacagctgtt
6001 tgtggaacag cacaagcact acctggacga gatcatcgag cagatcagcg agttctccaa
6061 gagagtgatc ctggccgacg ctaatctgga caaagtgctg tccgcctaca acaagcaccg
6121 ggataagccc atcagagagc aggccgagaa tatcatccac ctgtttaccc tgaccaatct
6181 gggagcccct gccgccttca agtactttga caccaccatc gaccggaaga ggtacaccag
6241 caccaaagag gtgctggacg ccaccctgat ccaccagagc atcaccggcc tgtacgagac
6301 acggatcgac ctgtctcagc tgggaggcga caagcgtcct gctgctacta agaaagctgg
6361 tcaagctaag aaaaagaaag ctagcggcag cggcgccacc aacttcagcc tgctgaagca
6421 ggccggcgac gtggaggaga accccggccc catgaccgag tacaagccca cggtgcgcct
6481 cgccacccgc gacgacgtcc ccagggccgt acgcaccctc gccgccgcgt tcgccgacta
6541 ccccgccacg cgccacaccg tcgatccgga ccgccacatc gagcgggtca ccgagctgca
6601 agaactcttc ctcacgcgcg tcgggctcga catcggcaag gtgtgggtcg cggacgacgg
6661 cgccgcggtg gcggtctgga ccacgccgga gagcgtcgaa gcgggggcgg tgttcgccga
6721 gatcggcccg cgcatggccg agttgagcgg ttcccggctg gccgcgcagc aacagatgga
6781 aggcctcctg gcgccgcacc ggcccaagga gcccgcgtgg ttcctggcca ccgtcggagt
6841 ctcgcccgac caccagggca agggtctggg cagcgccgtc gtgctccccg gagtggaggc
6901 ggccgagcgc gccggggtgc ccgccttcct ggagacctcc gcgccccgca acctcccctt
6961 ctacgagcgg ctcggcttca ccgtcaccgc cgacgtcgag gtgcccgaag gaccgcgcac
7021 ctggtgcatg acccgcaagc ccggtgcctg aacgcgttaa gtcgacaatc aacctctgga
7081 ttacaaaatt tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg
7141 tggatacgct gctttaatgc ctttgtatca tgctattgct tcccgtatgg ctttcatttt
7201 ctcctccttg tataaatcct ggttgctgtc tctttatgag gagttgtggc ccgttgtcag
7261 gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc cccactggtt ggggcattgc
7321 caccacctgt cagctccttt ccgggacttt cgctttcccc ctccctattg ccacggcgga
7381 actcatcgcc gcctgccttg cccgctgctg gacaggggct cggctgttgg gcactgacaa
7441 ttccgtggtg ttgtcgggga aatcatcgtc ctttccttgg ctgctcgcct gtgttgccac
7501 ctggattctg cgcgggacgt ccttctgcta cgtcccttcg gccctcaatc cagcggacct
7561 tccttcccgc ggcctgctgc cggctctgcg gcctcttccg cgtcttcgcc ttcgccctca
7621 gacgagtcgg atctcccttt gggccgcctc cccgcgtcga ctttaagacc aatgacttac
7681 aaggcagctg tagatcttag ccacttttta aaagaaaagg ggggactgga agggctaatt
7741 cactcccaac gaagacaaga tctgcttttt gcttgtactg ggtctctctg gttagaccag
7801 atctgagcct gggagctctc tggctaacta gggaacccac tgcttaagcc tcaataaagc
7861 ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt gtgactctgg taactagaga
7921 tccctcagac ccttttagtc agtgtggaaa atctctagca gtacgtatag tagttcatgt
7981 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
8041 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
8101 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
8161 catgtctggc tctagctatc ccgcccctaa ctccgcccat cccgccccta actccgccca
8221 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
8281 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggga
8341 cgtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg tcgttttaca
8401 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
8461 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
8521 cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
8581 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
8641 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
8701 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
8761 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
8821 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
8881 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
8941 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt
9001 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
9061 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
9121 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
9181 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
9241 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
9301 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
9361 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
9421 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
9481 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
9541 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
9601 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
9661 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
9721 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
9781 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
9841 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
9901 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
9961 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
10021 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
10081 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
10141 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
10201 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
10261 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
10321 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
10381 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
10441 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
10501 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
10561 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
10621 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
10681 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
10741 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
10801 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
10861 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
10921 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
10981 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
11041 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
11101 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
11161 aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg agc
//
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