| 质粒名称: | Cas9/pTREX-n |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 68708 |
| 存储实验室: | Roberto Docampo |
|
载体骨架基本信息 |
|
| 载体名称: | pTREX-n |
| 载体出品公司: | Vazquez and Levin., 1999 |
| 空载体大小: | 6227 bp |
| 完整质粒大小: | 11173 bp |
| 载体类型: | CRISPR |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | 氨苄霉素 |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 高拷贝 |
|
插入基因信息 |
|
| 插入基因名称: | Fusion gene Cas9-HA-2xNLS-GFP from vector pMJ920 、Cas9-GFP |
| 物种来源: | Synthetic; Streptococcus pyogenes |
| 插入大小: | 4975 bp |
| 基因突变: | C4239T mutation that eliminates a BamHI restrir |
| 标签/融合蛋白: | Fusion gene Cas9-HA-2xNLS-GFP from vector pMJ920 (Addgene) |
|
文献引用方法 |
|
| 材料和方法部分: | Cas9/pTREX-n was a gift from Roberto Docampo (Addgene plasmid # 68708 ; http://n2t.net/addgene:68708 ; RRID:Addgene_68708) |
| 参考文献部分: | CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2 Genes in Trypanosoma cruzi Reveals Their Role in Flagellar Attachment. Lander N, Li ZH, Niyogi S, Docampo R. MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15. 10.1128/mBio.01012-15 PubMed 26199333 |
买家导航
Cas9/pTREX-n载体质粒基本信息
Cas9/pTREX-n质粒图谱载体图谱和Cas9/pTREX-n载体序列质粒序列多克隆位点信息
Cas9/pTREX-n质粒载体简介
Cas9/pTREX-n质粒序列载体序列
LOCUS Cas9/pTREX-n 11171 bp DNA circular SYN 14-APR-2025
DEFINITION Expresses the fusion gene CAS9-HA-2xNLS-GFP in the pTREX-n backbone.
This vector is used for cloning a specific sgRNA by BamHI, to be
co-expressed with Cas9 for genome editing in Trypanosoma cruzi..
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 11171)
AUTHORS Lander N, Li ZH, Niyogi S, Docampo R
TITLE CRISPR/Cas9-Induced Disruption of Paraflagellar Rod Protein 1 and 2
Genes in Trypanosoma cruzi Reveals Their Role in Flagellar
Attachment.
JOURNAL MBio. 2015 Jul 21;6(4). pii: e01012-15. doi: 10.1128/mBio.01012-15.
PUBMED 26199333
REFERENCE 2 (bases 1 to 11171)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Apr 14, 2025 from SnapGene Server 8.0.1
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "MBio.";
date: "2015-07-21"; volume: "6(4). pii: e01012-15. doi"; pages: "
10.1128/mBio.01012-15"
COMMENT Sequence Label: Cas9/pTREX-n
FEATURES Location/Qualifiers
source 1..11171
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(33..461)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(143..164)
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind 355..374
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 588..610
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 602..619
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 603..619
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 626..645
/label=T7
/note="T7 promoter, forward primer"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 1690..5793
/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 1690..5793
/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 5797..5823
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
CDS 5803..5829
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/note="This is a variant of the standard HA tag, which has
sequence YPYDVPDYA. Some anti-HA antibodies may not
recognize this variant."
/translation="YDVPDYASL"
CDS 5842..5862
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 5869..5889
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS join(5929..5931,5932..6645)
/codon_start=1
/product="enhanced GFP (EGFP) derivative that exhibits
robust folding and enhanced photostability (Zhong et al.,
2019)"
/label=SiriusGFP
/note="mammalian codon-optimized"
/translation="VSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
FVTAAGITLGMDELYK"
primer_bind 6579..6600
/label=EGFP-C
/note="EGFP, forward primer"
primer_bind complement(6654..6670)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6655..6671)
/label=pBluescriptKS
/note="For pBluescript vector"
CDS 7408..8211
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MGSAIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQ
GRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQD
LLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDE
EHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQ
DIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(7471..7490)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 8081..8100
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
promoter complement(8986..9004)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(9025..9041)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9025..9041)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9038..9060)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9049..9065
/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(join(9073..9079,9080..9097,9098..9103))
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 9118..9139
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(9256..9273)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(9427..10015)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(9507..9526)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(join(10186..10977,10978..11046))
/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 10809..10828
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(11047..11151)
/gene="bla"
/label=AmpR promoter
ORIGIN
1 cacctaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
61 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
121 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
181 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
241 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
301 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
361 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
421 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcgg
661 tgcaccctgg gatatttgcg cacccacctt tcccgtcgca atcggcgccg tccgaacgcg
721 gaaatgttcc gagaataata cggctgggct gcgccgcgcg ttccccgcgt gaaccaacta
781 ttgtgcgatg tgcagcatcc tattgtggtg ctttaccgca caccagtttc cttgcggggc
841 ttatttggtg tgttggattt ttaatttttt gatgattcgg aggaattttt tgcaattttt
901 gggtacccct aaaaaaattt tggagtttcc ggggttcgat ccaacttgtg cttttttaac
961 cgtccatttg tgtctagtac atcatggagc ggtattctct ccggtggccc aggtggcgtc
1021 ggaaggaatc ggtgtccctc aggatgtttg gtttttcgtt tttttttttg cggtcttttg
1081 tttcgcatcg ccgtatcgtg tgcggcgcga ccttcttttt ggctcattgg tcagtgaggt
1141 ttataataaa gtggatcatt tgggtttttg gccctttgcg gcgcacccgc ggaatggggg
1201 agggattttc atataattgt agtgtacaca ttgtgtgtgg gtcgcaaaat gtcatgccgt
1261 gtgcaatgtt tatttgcgtg ctgtgcgtgt ccttttgtga ttccaccact tgtgtgtatt
1321 ttaccttatc gtctctccct gccgttggct gtgcgtgaat tgtgcttcgc attcgtggga
1381 aatcaaaagg ggaatcaacg tgttcttttt ttttcagaag gggagtcaaa gcgccgtgtg
1441 gatgccaagt ctgtgcaaga gttctagtgg atcccccaac gagtttcttc aaaatatgca
1501 gcggattcac taagaaacat tttcacgcac gaaagcgaaa ttattatgat tgttattata
1561 atactttttc tttgttgttt tatccactta ttattgttgt gttaaattgt ttttaccttt
1621 tttcttttcc aacttctttt atgatgtctt ttcttttttt tttttgctct ataagttgtc
1681 ttgtctagaa tggacaagaa gtacagcatc ggcctggaca tcggcaccaa ctctgtgggc
1741 tgggccgtga tcaccgacga gtacaaggtg cccagcaaga aattcaaggt gctgggcaac
1801 accgaccggc acagcatcaa gaagaacctg atcggcgccc tgctgttcga cagcggagaa
1861 acagccgagg ccacccggct gaagagaacc gccagaagaa gatacaccag acggaagaac
1921 cggatctgct atctgcaaga gatcttcagc aacgagatgg ccaaggtgga cgacagcttc
1981 ttccacagac tggaagagtc cttcctggtg gaagaggata agaagcacga gcggcacccc
2041 atcttcggca acatcgtgga cgaggtggcc taccacgaga agtaccccac catctaccac
2101 ctgagaaaga aactggtgga cagcaccgac aaggccgacc tgcggctgat ctatctggcc
2161 ctggcccaca tgatcaagtt ccggggccac ttcctgatcg agggcgacct gaaccccgac
2221 aacagcgacg tggacaagct gttcatccag ctggtgcaga cctacaacca gctgttcgag
2281 gaaaacccca tcaacgccag cggcgtggac gccaaggcca tcctgtctgc cagactgagc
2341 aagagcagac ggctggaaaa tctgatcgcc cagctgcccg gcgagaagaa gaatggcctg
2401 ttcggcaacc tgattgccct gagcctgggc ctgaccccca acttcaagag caacttcgac
2461 ctggccgagg atgccaaact gcagctgagc aaggacacct acgacgacga cctggacaac
2521 ctgctggccc agatcggcga ccagtacgcc gacctgtttc tggccgccaa gaacctgtcc
2581 gacgccatcc tgctgagcga catcctgaga gtgaacaccg agatcaccaa ggcccccctg
2641 agcgcctcta tgatcaagag atacgacgag caccaccagg acctgaccct gctgaaagct
2701 ctcgtgcggc agcagctgcc tgagaagtac aaagagattt tcttcgacca gagcaagaac
2761 ggctacgccg gctacatcga tggcggagcc agccaggaag agttctacaa gttcatcaag
2821 cccatcctgg aaaagatgga cggcaccgag gaactgctcg tgaagctgaa cagagaggac
2881 ctgctgcgga agcagcggac cttcgacaac ggcagcatcc cccaccagat ccacctggga
2941 gagctgcacg ccattctgcg gcggcaggaa gatttttacc cattcctgaa ggacaaccgg
3001 gaaaagatcg agaagatcct gaccttccgc atcccctact acgtgggccc tctggccagg
3061 ggaaacagca gattcgcctg gatgaccaga aagagcgagg aaaccatcac cccctggaac
3121 ttcgaggaag tggtggacaa gggcgccagc gcccagagct tcatcgagcg gatgaccaac
3181 ttcgataaga acctgcccaa cgagaaggtg ctgcccaagc acagcctgct gtacgagtac
3241 ttcaccgtgt acaacgagct gaccaaagtg aaatacgtga ccgagggaat gagaaagccc
3301 gccttcctga gcggcgagca gaaaaaagcc atcgtggacc tgctgttcaa gaccaaccgg
3361 aaagtgaccg tgaagcagct gaaagaggac tacttcaaga aaatcgagtg cttcgactcc
3421 gtggaaatct ccggcgtgga agatcggttc aacgcctccc tgggcacata ccacgatctg
3481 ctgaaaatta tcaaggacaa ggacttcctg gacaatgagg aaaacgagga cattctggaa
3541 gatatcgtgc tgaccctgac actgtttgag gacagagaga tgatcgagga acggctgaaa
3601 acctatgccc acctgttcga cgacaaagtg atgaagcagc tgaagcggcg gagatacacc
3661 ggctggggca ggctgagccg gaagctgatc aacggcatcc gggacaagca gtccggcaag
3721 acaatcctgg atttcctgaa gtccgacggc ttcgccaaca gaaacttcat gcagctgatc
3781 cacgacgaca gcctgacctt taaagaggac atccagaaag cccaggtgtc cggccagggc
3841 gatagcctgc acgagcacat tgccaatctg gccggcagcc ccgccattaa gaagggcatc
3901 ctgcagacag tgaaggtggt ggacgagctc gtgaaagtga tgggccggca caagcccgag
3961 aacatcgtga tcgaaatggc cagagagaac cagaccaccc agaagggaca gaagaacagc
4021 cgcgagagaa tgaagcggat cgaagagggc atcaaagagc tgggcagcca gatcctgaaa
4081 gaacaccccg tggaaaacac ccagctgcag aacgagaagc tgtacctgta ctacctgcag
4141 aatgggcggg atatgtacgt ggaccaggaa ctggacatca accggctgtc cgactacgat
4201 gtggaccata tcgtgcctca gagctttctg aaggacgact ccatcgataa caaagtgctg
4261 actcggagcg acaagaaccg gggcaagagc gacaacgtgc cctccgaaga ggtcgtgaag
4321 aagatgaaga actactggcg ccagctgctg aatgccaagc tgattaccca gaggaagttc
4381 gacaatctga ccaaggccga gagaggcggc ctgagcgaac tggataaggc cggcttcatc
4441 aagagacagc tggtggaaac ccggcagatc acaaagcacg tggcacagat cctggactcc
4501 cggatgaaca ctaagtacga cgagaacgac aaactgatcc gggaagtgaa agtgatcacc
4561 ctgaagtcca agctggtgtc cgatttccgg aaggatttcc agttttacaa agtgcgcgag
4621 atcaacaact accaccacgc ccacgacgcc tacctgaacg ccgtcgtggg aaccgccctg
4681 atcaaaaagt accctaagct ggaaagcgag ttcgtgtacg gcgactacaa ggtgtacgac
4741 gtgcggaaga tgatcgccaa gagcgagcag gaaatcggca aggctaccgc caagtacttc
4801 ttctacagca acatcatgaa ctttttcaag accgagatta ccctggccaa cggcgagatc
4861 cggaagcggc ctctgatcga gacaaacggc gaaacaggcg agatcgtgtg ggataagggc
4921 cgggactttg ccaccgtgcg gaaagtgctg tctatgcccc aagtgaatat cgtgaaaaag
4981 accgaggtgc agacaggcgg cttcagcaaa gagtctatcc tgcccaagag gaacagcgac
5041 aagctgatcg ccagaaagaa ggactgggac cctaagaagt acggcggctt cgacagcccc
5101 accgtggcct attctgtgct ggtggtggcc aaagtggaaa agggcaagtc caagaaactg
5161 aagagtgtga aagagctgct ggggatcacc atcatggaaa gaagcagctt cgagaagaat
5221 cccatcgact ttctggaagc caagggctac aaagaagtga aaaaggacct gatcatcaag
5281 ctgcctaagt actccctgtt cgagctggaa aacggccgga agagaatgct ggcctctgcc
5341 ggcgaactgc agaagggaaa cgaactggcc ctgccctcca aatatgtgaa cttcctgtac
5401 ctggccagcc actatgagaa gctgaagggc tcccccgagg ataatgagca gaaacagctg
5461 tttgtggaac agcacaaaca ctacctggac gagatcatcg agcagatcag cgagttctcc
5521 aagagagtga tcctggccga cgctaatctg gacaaggtgc tgagcgccta caacaagcac
5581 agagacaagc ctatcagaga gcaggccgag aatatcatcc acctgtttac cctgaccaat
5641 ctgggagccc ctgccgcctt caagtacttt gacaccacca tcgaccggaa gaggtacacc
5701 agcaccaaag aggtgctgga cgccaccctg atccaccaga gcatcaccgg cctgtacgag
5761 acacggatcg acctgtctca gctgggaggc gacgcctatc cctatgacgt gcccgattat
5821 gccagcctgg gcagcggctc ccccaagaaa aaacgcaagg tggaagatcc taagaaaaag
5881 cggaaagtgg acggcattgg tagtgggagc aacggcagca gcggatctgt gagcaagggc
5941 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
6001 cacaagttca gcgtgcgcgg cgagggcgag ggcgatgcca ccaacggcaa gctgaccctg
6061 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
6121 acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
6181 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tctccttcaa ggacgacggc
6241 acctacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
6301 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
6361 ttcaacagcc acaacgtcta tatcacggcc gacaagcaga agaacggcat caaggcgaac
6421 ttcaagatcc gccacaacgt cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
6481 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
6541 tccaagctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
6601 accgccgccg ggatcactct cggcatggac gagctgtaca agtagaagct tatcgatacc
6661 gtcgacctcg agccatttac gactccaagg caacgctgca gaacaacctg ccgaaggagc
6721 gccgcttctt caagattgtg tcgtggtacg acaacgagtg gggatactcc caccgcgtgg
6781 tggaccttgt acgccacatg gcctcgaagg atcgttcggc aaggttgtag gcgtggcgat
6841 gacttcaggt ctttcttttg cgaataggga tcttataata cacgatgcgt gtcccgtgat
6901 gatcgttacc ggtgctgcca cgatccaatt gacacagcgt caagagcaaa acaattttac
6961 ttttcccttt aaggacaaca acaaaaaaat atataacttt tttttctttt tttttttttt
7021 ttgaaattat atttatggtc atctttggga aacaaaaagc agcaatttaa tgatgcggaa
7081 ggatgagtga aataatgttt aatcaatgta cgaggatttg gggtattgca aggaaaatgt
7141 agatgattta attgggtgtg tgatgcagct tgtggtaatt tttgctcact tccctttttg
7201 ccacatcttt ttagtttttc tgcttttctt tccccattat tccacttgtc tctcttttcc
7261 cacgtttcct gcacgaatgc agaaagtgat atttttactt tgaaagccat ctaccaacaa
7321 caattacatt gaacagaatt tgggattgcg aattaattcc tgcagccccg cgtgtggcct
7381 cgaacaccga gcgaccctgc agccaatatg ggatcggcca ttgaacaaga tggattgcac
7441 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
7501 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
7561 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
7621 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
7681 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
7741 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
7801 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
7861 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
7921 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
7981 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
8041 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
8101 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
8161 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg aggggatcga
8221 tccggaacaa cctgccgaag gagcgccgct tcttcaagat tgtgtcgtgg tacgacaacg
8281 agtggggata ctcccaccgc gtggtggacc ttgtacgcca catggcctcg aaggatcgtt
8341 cggcaaggtt gtaggcgtgg cgatgacttc aggtctttct tttgcgaata gggatcttat
8401 aatacacgat gcgtgtcccg tgatgatcgt taccggtgct gccacgatcc aagtgacaca
8461 gtgtcaagag caaaacaatt ttacttttcc ctttaaggac aacaacaaaa aaatatataa
8521 cttttttttc tttttttttt tttgaaatta tatttatggt catctttggg aaacaaaaag
8581 cagcaattta atgatgcgga aggatgagtg aaataatgtt taatcaatgt acgaggattt
8641 ggggtattgc aaggaaaatg tagatgattt agttgggtgt atgatgcagc ttgtggtaat
8701 tttttacaac tttgcgttat tggaagattt tggttgttca gtttcttttt gtttataact
8761 gactgtttta atttagtgtg attttttttt tttaaggatt taagctgttt ttcttttttt
8821 ttctttttct tcactgatgg tgtttaaggt gtagtttagt gtttgtgcat tatgcgtcgt
8881 agtgttgttg ttctttggac tttttcgaac aatactgggc attatagccc acctgtgtat
8941 atgccgctag agtatgctag ccgtgtgggt acccagcttt tgttcccttt agtgagggtt
9001 aatttcgagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
9061 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
9121 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
9181 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
9241 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
9301 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
9361 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
9421 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
9481 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
9541 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
9601 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
9661 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
9721 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
9781 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
9841 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
9901 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
9961 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
10021 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
10081 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
10141 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
10201 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
10261 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
10321 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
10381 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
10441 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
10501 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
10561 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
10621 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
10681 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
10741 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
10801 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
10861 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
10921 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
10981 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
11041 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
11101 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
11161 ccgaaaagtg c
//
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