| 质粒名称: | pCasAb-apr |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 121998 |
| 存储实验室: | Quanjiang Ji |
|
载体骨架基本信息 |
|
| 载体名称: | pMMB67EH |
| 载体出品公司: | Clontech |
| 空载体大小: | 16599 bp |
| 完整质粒大小: | 16600 bp |
| 载体类型: | Bacterial Expression, CRISPR |
| 筛选标记: | 新霉素（G418） |
| 载体抗性: | Apramycin, 25 μg/mL |
| 生长温度: | 37 ℃ |
| 宿主菌: | DH5a |
| 拷贝数: | 低拷贝 |
|
插入基因信息 |
|
| 克隆方法: | Gibson Cloning |
|
文献引用方法 |
|
| 材料和方法部分: | pCasAb-apr was a gift from Quanjiang Ji (Addgene plasmid # 121998 ; http://n2t.net/addgene:121998 ; RRID:Addgene_121998) |
| 参考文献部分: | A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of OxyR. Wang Y, Wang Z, Chen Y, Hua X, Yu Y, Ji Q. Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi: 10.1016/j.chembiol.2019.09.003. 10.1016/j.chembiol.2019.09.003 PubMed 31548010 |
买家导航
pCasAb-apr载体质粒基本信息
pCasAb-apr质粒图谱载体图谱和pCasAb-apr载体序列质粒序列多克隆位点信息
pCasAb-apr质粒载体简介
pCasAb-apr质粒序列载体序列
LOCUS pCasAb-apr 16600 bp DNA circular SYN 14-APR-2025
DEFINITION Bacterial expression of Cas9 nuclease and RecAb recombination system
in Acinetobacter baumannii.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 16600)
AUTHORS Wang Y, Wang Z, Chen Y, Hua X, Yu Y, Ji Q
TITLE A Highly Efficient CRISPR-Cas9-Based Genome Engineering Platform in
Acinetobacter baumannii to Understand the H2O2-Sensing Mechanism of
OxyR.
JOURNAL Cell Chem Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi:
10.1016/j.chembiol.2019.09.003.
PUBMED 31548010
REFERENCE 2 (bases 1 to 16600)
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: "Cell Chem
Biol. 2019 Sep 17. pii: S2451-9456(19)30277-6. doi:
10.1016/j.chembiol.2019.09.003."
COMMENT Sequence Label: pCasAb-apr
FEATURES Location/Qualifiers
source 1..16600
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(38..55)
/label=pBAD Reverse
/note="For vectors with E. coli araBAD promoter, reverse
primer"
terminator 208..294
/gene="Escherichia coli rrnB"
/label=rrnB T1 terminator
/note="transcription terminator T1 from the E. coli rrnB
gene"
terminator 386..413
/label=rrnB T2 terminator
/note="transcription terminator T2 from the E. coli rrnB
gene"
promoter join(678..683,684..699,700..706)
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 714..730
/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)."
CDS 753..4859
/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 753..4856
/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 join(5271..5357,5358..6692)
/codon_start=1
/gene="Bacillus subtilis sacB"
/product="secreted levansucrase that renders bacterial
growth sensitive to sucrose"
/label=SacB
/note="negative selection marker"
/translation="MNIKKFAKQATVLTFTTALLAGGATQAFAKETNQKPYKETYGISH
ITRHDMLQIPEQQKNEKYQVPEFDSSTIKNISSAKGLDVWDSWPLQNADGTVANYHGYH
IVFALAGDPKNADDTSIYMFYQKVGETSIDSWKNAGRVFKDSDKFDANDSILKDQTQEW
SGSATFTSDGKIRLFYTDFSGKHYGKQTLTTAQVNVSASDSSLNINGVEDYKSIFDGDG
KTYQNVQQFIDEGNYSSGDNHTLRDPHYVEDKGHKYLVFEANTGTEDGYQGEESLFNKA
YYGKSTSFFRQESQKLLQSDKKRTAELANGALGMIELNDDYTLKKVMKPLIASNTVTDE
IERANVFKMNGKWYLFTDSRGSKMTIDGITSNDIYMLGYVSNSLTGPYKPLNKTGLVLK
MDLDPNDVTFTYSHFAVPQAKGNNVVITSYMTNRGFYADKQSTFAPSFLLNIKGKKTSV
VKDSILEQGQLTVNK"
primer_bind complement(6731..6750)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind complement(6754..6770)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(6755..6771)
/label=pBluescriptKS
/note="For pBluescript vector"
CDS 6909..7709
/codon_start=1
/gene="aac(3)IV"
/product="aminoglycoside 3-N-acetyltransferase type IV
"
/label=ApmR
/note="confers resistance to apramycin (Paget and Davies,
1996)"
/translation="MSSAVECNVVQYEWRKAELIGQLLNLGVTPGGVLLVHSSFRSVRP
LEDGPLGLIEALRAALGPGGTLVMPSWSGLDDEPFDPATSPVTPDLGVVSDTFWRLPNV
KRSAHPFAFAAAGPQAEQIISDPLPLPPHSPASPVARVHELDGQVLLLGVGHDANTTLH
LAELMAKVPYGVPRHCTILQDGKLVRVDYLENDHCCERFALADRWLKEKSLQKEGPVGH
AFARLIRSRDIVATALGQLGRDPLIFLHPPEAGCEECDAARQSIG"
rep_origin 8149..8543
/label=RSF1010 oriV
/note="replication origin of the broad-host-range plasmid
RSF1010; requires the RSF1010 RepA/B/C proteins for
replication (Scholz et al., 1989)"
oriT 8885..8972
/label=RSF1010 oriT
/note="origin of transfer of the broad-host-range plasmid
RSF1010 (Scholz et al., 1989)"
CDS 10211..11182
/codon_start=1
/gene="repB"
/product="replication protein B of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepB
/translation="MKNDRTLQAIGRQLKAMGCERFDIGVRDATTGQMMNREWSAAEVL
QNTPWLKRMNAQGNDVYIRPAEQERHGLVLVDDLSEFDLDDMKAEGREPALVVETSPKN
YQAWVKVADAAGGELRGQIARTLASEYDADPASADSRHYGRLAGFTNRKDKHTTRAGYQ
PWVLLRESKGKTATAGPALVQQAGQQIEQAQRQQEKARRLASLELPERQLSRHRRTALD
EYRSEMAGLVKRFGDDLSKCDFIAAQKLASRGRSAEEIGKAMAEASPALAERKPGHEAD
YIERTVSKVMGLPSVQLARAELARAPAPRQRGMDRGGPDFSM"
CDS 11693..12532
/codon_start=1
/gene="repA"
/product="replication protein A of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepA
/translation="MATHKPINILEAFAAAPPPLDYVLPNMVAGTVGALVSPGGAGKSM
LALQLAAQIAGGPDLLEVGELPTGPVIYLPAEDPPTAIHHRLHALGAHLSAEERQAVAD
GLLIQPLIGSLPNIMAPEWFDGLKRAAEGRRLMVLDTLRRFHIEEENASGPMAQVIGRM
EAIAADTGCSIVFLHHASKGAAMMGAGDQQQASRGSSVLVDNIRWQSYLSSMTSAEAEE
WGVDDDQRRFFVRFGVSKANYGAPFADRWFRRHDGGVLKPAVLERQRKSKGVPRGEA"
CDS 12519..13370
/codon_start=1
/gene="repC"
/product="replication protein C of the broad-host-range
plasmid RSF1010 (Scholz et al., 1989)"
/label=RSF1010 RepC
/translation="MVKPKNKHSLSHVRHDPAHCLAPGLFRALKRGERKRSKLDVTYDY
GDGKRIEFSGPEPLGADDLRILQGLVAMAGPNGLVLGPEPKTEGGRQLRLFLEPKWEAV
TADAMVVKGSYRALAKEIGAEVDSGGALKHIQDCIERLWKVSIIAQNGRKRQGFRLLSE
YASDEADGRLYVALNPLIAQAVMGGGQHVRISMDEVRALDSETARLLHQRLCGWIDPGK
TGKASIDTLCGYVWPSEASGSTMRKRRQRVREALPELVALGWTVTEFAAGKYDITRPKA
AG"
CDS complement(13647..14729)
/codon_start=1
/gene="lacI"
/product="lac repressor"
/label=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)."
/translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
ALADSLMQLARQVSRLESGQ"
primer_bind 14691..14710
/label=LacI-R
/note="LacI, reverse primer"
promoter complement(14730..14807)
/gene="lacI (mutant)"
/label=lacIq promoter
/note="In the lacIq allele, a single base change in the
promoter boosts expression of the lacI gene about 10-fold."
promoter join(15037..15042,15043..15058,15059..15065)
/label=tac promoter
/note="strong E. coli promoter; hybrid between the trp and
lac UV5 promoters"
protein_bind 15073..15089
/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)."
ORIGIN
1 agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt aaatcagaac
61 gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg gtcccacctg
121 accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg gggtctcccc
181 atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
241 gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac aaatccgccg
301 ggagcggatt tgaacgttgc gaagcaacgg cccggagggt ggcgggcagg acgcccgcca
361 taaactgcca ggcatcaaat taagcagaag gccatcctga cggatggcct ttttgcgttt
421 ctacaaactc ttttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
481 ataaccctgg atccgggctt atcgactgca cggtgcacca atgcttctgg cgtcaggcag
541 ccatcggaag ctgtggtatg gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc
601 aaggcgcact cccgttctgg ataatgtttt ttgcgccgac atcataacgg ttctggcaaa
661 tattctgaaa tgagctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
721 cggataacaa tttcacacag gaaacagaat tcatggataa gaaatactca ataggcttag
781 atatcggcac aaatagcgtc ggatgggcgg tgatcactga tgaatataag gttccgtcta
841 aaaagttcaa ggttctggga aatacagacc gccacagtat caaaaaaaat cttatagggg
901 ctcttttatt tgacagtgga gagacagcgg aagcgactcg tctcaaacgg acagctcgta
961 gaaggtatac acgtcggaag aatcgtattt gttatctaca ggagattttt tcaaatgaga
1021 tggcgaaagt agatgatagt ttctttcatc gacttgaaga gtcttttttg gtggaagaag
1081 acaagaagca tgaacgtcat cctatttttg gaaatatagt agatgaagtt gcttatcatg
1141 agaaatatcc aactatctat catctgcgaa aaaaattggt agattctact gataaagcgg
1201 atttgcgctt aatctatttg gccttagcgc atatgattaa gtttcgtggt cattttttga
1261 ttgagggaga tttaaatcct gataatagtg atgtggacaa actatttatc cagttggtac
1321 aaacctacaa tcaattattt gaagaaaacc ctattaacgc aagtggagta gatgctaaag
1381 cgattctttc tgcacgattg agtaaatcaa gacgattaga aaatctcatt gctcagctcc
1441 ccggtgagaa gaaaaatggc ttatttggga atctcattgc tttgtcattg ggtttgaccc
1501 ctaattttaa atcaaatttt gatttggcag aagatgctaa attacagctt tcaaaagata
1561 cttacgatga tgatttagat aatttattgg cgcaaattgg agatcaatat gctgatttgt
1621 ttttggcagc taagaattta tcagatgcta ttttactttc agatatccta agagtaaata
1681 ctgaaataac taaggctccc ctatcagctt caatgattaa acgctacgat gaacatcatc
1741 aagacttgac tcttttaaaa gctttagttc gacaacaact tccagaaaag tataaagaaa
1801 tcttttttga tcaatcaaaa aacggatatg caggttatat tgatggggga gctagccaag
1861 aagaatttta taaatttatc aaaccaattt tagaaaaaat ggatggtact gaggaattat
1921 tggtgaaact aaatcgtgaa gatttgctgc gcaagcaacg gacctttgac aacggctcta
1981 ttccccatca aattcacttg ggtgagctgc atgctatttt gagaagacaa gaagactttt
2041 atccattttt aaaagacaat cgtgagaaga ttgaaaaaat cttgactttt cgaattcctt
2101 attatgttgg tccattggcg cgtggcaata gtcgttttgc atggatgact cggaagtctg
2161 aagaaacaat taccccatgg aattttgaag aagttgtcga taaaggtgct tcagctcaat
2221 catttattga acgcatgaca aactttgata aaaatcttcc aaatgaaaaa gtactaccaa
2281 aacatagttt gctttatgag tattttacgg tttataacga attgacaaag gtcaaatatg
2341 ttactgaagg aatgcgaaaa ccagcatttc tttcaggtga acagaagaaa gccattgttg
2401 atttactctt caaaacaaat cgaaaagtaa ccgttaagca attaaaagaa gattatttca
2461 aaaaaataga atgttttgat agtgttgaaa tttcaggagt tgaagataga tttaatgctt
2521 cattaggtac ctaccatgat ttgctaaaaa ttattaaaga taaagatttt ttggataatg
2581 aagaaaatga agatatctta gaggatattg ttttaacatt gaccttattt gaagataggg
2641 agatgattga ggaaagactt aaaacatatg ctcacctctt tgatgataag gtgatgaaac
2701 agcttaaacg tcgccgttat actggttggg gacgtttgtc tcgaaaattg attaatggta
2761 ttagggataa gcaatctggc aaaacaatat tagatttttt gaaatcagat ggttttgcca
2821 atcgcaattt tatgcagctg atccatgatg atagtttgac atttaaagaa gacattcaaa
2881 aagcacaagt gtctggacaa ggcgatagtt tacatgaaca tattgcaaat ttagctggta
2941 gccctgctat taaaaaaggt attttacaga ctgtaaaagt tgttgatgaa ttggtcaaag
3001 taatggggcg gcataagcca gaaaatatcg ttattgaaat ggcacgtgaa aatcagacaa
3061 ctcaaaaggg ccagaaaaat tcgcgagagc gtatgaaacg aatcgaagaa ggtatcaaag
3121 aattaggaag tcagattctt aaagagcatc ctgttgaaaa tactcaattg caaaatgaaa
3181 agctctatct ctattatctc caaaatggaa gagacatgta tgtggaccaa gaattagata
3241 ttaatcgttt aagtgattat gatgtcgatc acattgttcc acaaagtttc cttaaagacg
3301 attcaataga caataaggtc ttaacgcgtt ctgataaaaa tcgtggtaaa tcggataacg
3361 ttccaagtga agaagtagtc aaaaagatga aaaactattg gagacaactt ctaaacgcca
3421 agttaatcac tcaacgtaag tttgataatt taacgaaagc tgaacgtgga ggtttgagtg
3481 aacttgataa agctggtttt atcaaacgcc aattggttga aactcgccaa atcactaagc
3541 atgtggcaca aattttggat agtcgcatga atactaaata cgatgaaaat gataaactta
3601 ttcgagaggt taaagtgatt accttaaaat ctaaattagt ttctgacttc cgaaaagatt
3661 tccaattcta taaagtacgt gagattaaca attaccatca tgcccatgat gcgtatctaa
3721 atgccgtcgt tggaactgct ttgattaaga aatatccaaa acttgaatcg gagtttgtct
3781 atggtgatta taaagtttat gatgttcgta aaatgattgc taagtctgag caagaaatag
3841 gcaaagcaac cgcaaaatat ttcttttact ctaatatcat gaacttcttc aaaacagaaa
3901 ttacacttgc aaatggagag attcgcaaac gccctctaat cgaaactaat ggggaaactg
3961 gagaaattgt ctgggataaa gggcgagatt ttgccacagt gcgcaaagta ttgtccatgc
4021 cccaagtcaa tattgtcaag aaaacagaag tacagacagg cggattctcc aaggagtcaa
4081 ttttaccaaa aagaaattcg gacaagctta ttgctcgtaa aaaagactgg gatccaaaaa
4141 aatatggtgg ttttgatagt ccaacggtag cttattcagt cctagtggtt gctaaggtgg
4201 aaaaagggaa atcgaagaag ttaaaatccg ttaaagagtt actagggatc acaattatgg
4261 aaagaagttc ctttgaaaaa aatccgattg actttttaga agctaaagga tataaggaag
4321 ttaaaaaaga cttaatcatt aaactaccta aatatagtct ttttgagtta gaaaacggtc
4381 gtaaacggat gctggctagt gccggagaat tacaaaaagg aaatgagctg gctctgccaa
4441 gcaaatatgt gaatttttta tatttagcta gtcattatga aaagttgaag ggtagtccag
4501 aagataacga acaaaaacaa ttgtttgtgg agcagcataa gcattattta gatgagatta
4561 ttgagcaaat cagtgaattt tctaagcgtg ttattttagc agatgccaat ttagataaag
4621 ttcttagtgc atataacaaa catagagaca aaccaatacg tgaacaagca gaaaatatta
4681 ttcatttatt tacgttgacg aatcttggag ctcccgctgc ttttaaatat tttgatacaa
4741 caattgatcg taaacgatat acgtctacaa aagaagtttt agatgccact cttatccatc
4801 aatccatcac tggtctttat gaaacacgca ttgatttgag tcagctagga ggtgactgat
4861 ggctggttgg cgtactgttg tggtaaatac caagcttgca actttatgcc catgcaacag
4921 aaactataaa aaatacagag aatgaaaaga aacagataga ttttttagtt ctttaggccc
4981 gtagtctgca aatcctttta tgattttcta tcaaacaaaa gaggaaaata gaccagttgc
5041 aatccaaacg agagtctaat agaatgaggt cgaaaagtaa atcgcgcggg tttgttactg
5101 ataaagcagg caagacctaa aatgtgtaaa gggcaaagtg tatactttgg cgtcacccct
5161 tacatatttt aggtcttttt ttattgtgcg taactaactt gccatcttca aacaggaggg
5221 ctggaagaag cagaccgcta acacagtaca taaaaaagga gacatgaacg atgaacatca
5281 aaaagtttgc aaaacaagca acagtattaa cctttactac cgcactgctg gcaggaggcg
5341 caactcaagc gtttgcgaaa gaaacgaacc aaaagccata taaggaaaca tacggcattt
5401 cccatattac acgccatgat atgctgcaaa tccctgaaca gcaaaaaaat gaaaaatatc
5461 aagttcctga gttcgattcg tccacaatta aaaatatctc ttctgcaaaa ggcctggacg
5521 tttgggacag ctggccatta caaaacgctg acggcactgt cgcaaactat cacggctacc
5581 acatcgtctt tgcattagcc ggagatccta aaaatgcgga tgacacatcg atttacatgt
5641 tctatcaaaa agtcggcgaa acttctattg acagctggaa aaacgctggc cgcgtcttta
5701 aagacagcga caaattcgat gcaaatgatt ctatcctaaa agaccaaaca caagaatggt
5761 caggttcagc cacatttaca tctgacggaa aaatccgttt attctacact gatttctccg
5821 gtaaacatta cggcaaacaa acactgacaa ctgcacaagt taacgtatca gcatcagaca
5881 gctctttgaa catcaacggt gtagaggatt ataaatcaat ctttgacggt gacggaaaaa
5941 cgtatcaaaa tgtacagcag ttcatcgatg aaggcaacta cagctcaggc gacaaccata
6001 cgctgagaga tcctcactac gtagaagata aaggccacaa atacttagta tttgaagcaa
6061 acactggaac tgaagatggc taccaaggcg aagaatcttt atttaacaaa gcatactatg
6121 gcaaaagcac atcattcttc cgtcaagaaa gtcaaaaact tctgcaaagc gataaaaaac
6181 gcacggctga gttagcaaac ggcgctctcg gtatgattga gctaaacgat gattacacac
6241 tgaaaaaagt gatgaaaccg ctgattgcat ctaacacagt aacagatgaa attgaacgcg
6301 cgaacgtctt taaaatgaac ggcaaatggt atctgttcac tgactcccgc ggatcaaaaa
6361 tgacgattga cggcattacg tctaacgata tttacatgct tggttatgtt tctaattctt
6421 taactggccc atacaagccg ctgaacaaaa ctggccttgt gttaaaaatg gatcttgatc
6481 ctaacgatgt aacctttact tactcacact tcgctgtacc tcaagcgaaa ggaaacaatg
6541 tcgtgattac aagctatatg acaaacagag gattctacgc agacaaacaa tcaacgtttg
6601 cgcctagctt cctgctgaac atcaaaggca agaaaacatc tgttgtcaaa gacagcatcc
6661 ttgaacaagg acaattaaca gttaacaaat aaaaacgcaa aagaaaatgc cgattatggt
6721 gcactctcag tacaatctgc tctgatgccg tatcgatacc gtcgacctcg agggggggcc
6781 cggtaccatc ccaatacgcg tcaattcacg gatccggttc atgtgcagct ccatcagcaa
6841 aaggggatga taagtttatc accaccgact atttgcaaca gtgccgttga tcgtgctatg
6901 atcgactgat gtcatcagcg gtggagtgca atgtcgtgca atacgaatgg cgaaaagccg
6961 agctcatcgg tcagcttctc aaccttgggg ttacccccgg cggtgtgctg ctggtccaca
7021 gctccttccg tagcgtccgg cccctcgaag atgggccact tggactgatc gaggccctgc
7081 gtgctgcgct gggtccggga gggacgctcg tcatgccctc gtggtcaggt ctggacgacg
7141 agccgttcga tcctgccacg tcgcccgtta caccggacct tggagttgtc tctgacacat
7201 tctggcgcct gccaaatgta aagcgcagcg cccatccatt tgcctttgcg gcagcggggc
7261 cacaggcaga gcagatcatc tctgatccat tgcccctgcc acctcactcg cctgcaagcc
7321 cggtcgcccg tgtccatgaa ctcgatgggc aggtacttct cctcggcgtg ggacacgatg
7381 ccaacacgac gctgcatctt gccgagttga tggcaaaggt tccctatggg gtgccgagac
7441 actgcaccat tcttcaggat ggcaagttgg tacgcgtcga ttatctcgag aatgaccact
7501 gctgtgagcg ctttgccttg gcggacaggt ggctcaagga gaagagcctt cagaaggaag
7561 gtccagtcgg tcatgccttt gctcggttga tccgctcccg cgacattgtg gcgacagccc
7621 tgggtcaact gggccgagat ccgttgatct tcctgcatcc gccagaggcg ggatgcgaag
7681 aatgcgatgc cgctcgccag tcgattggct gactgtcaga ccaagtttac tcatatatac
7741 tttagattga tttctgaaag cgaccaggtg ctcggcgtgg caagactcgc agcgaacccg
7801 tagaaagcca tgctccagcc gcccgcattg gagaaattct tcaaattccc gttgcacata
7861 gcccggcaat tcctttccct gctctgccat aagcgcagcg aatgccgggt aatactcgtc
7921 aacgatctga tagagaaggg tttgctcggg tcggtggctc tggtaacgac cagtatcccg
7981 atcccggctg gccgtcctgg ccgccacatg aggcatgttc cgcgtccttg caatactgtg
8041 tttacataca gtctatcgct tagcggaaag ttcttttacc ctcagccgaa atgcctgccg
8101 ttgctagaca ttgccagcca gtgcccgtca ctcccgtact aactgtcacg aacccctgca
8161 ataactgtca cgcccccctg caataactgt cacgaacccc tgcaataact gtcacgcccc
8221 caaacctgca aacccagcag gggcgggggc tggcggggtg ttggaaaaat ccatccatga
8281 ttatctaaga ataatccact aggcgcggtt atcagcgccc ttgtggggcg ctgctgccct
8341 tgcccaatat gcccggccag aggccggata gctggtctat tcgctgcgct aggctacaca
8401 ccgccccacc gctgcgcggc agggggaaag gcgggcaaag cccgctaaac cccacaccaa
8461 accccgcaga aatacgctgg agcgctttta gccgctttag cggcctttcc ccctacccga
8521 agggtggggg cgcgtgtgca gccccgcagg gcctgtctcg gtcgatcatt cagcccggct
8581 catccttctg gcgtggcggc agaccgaaca aggcgcggtc gtggtcgcgt tcaaggtacg
8641 catccattgc cgccatgagc cgatcctccg gccactcgct gctgttcacc ttggccaaaa
8701 tcatggcccc caccagcacc ttgcgccttg tttcgttctt gcgctcttgc tgctgttccc
8761 ttgcccgcac ccgctgaatt tcggcattga ttcgcgctcg ttgttcttcg agcttggcca
8821 gccgatccgc cgccttgttg ctccccttaa ccatcttgac accccattgt taatgtgctg
8881 tctcgtaggc tatcatggag gcacagcggc ggcaatcccg accctacttt gtaggggagg
8941 gcgcacttac cggtttctct tcgagaaact ggcctaacgg ccacccttcg ggcggtgcgc
9001 tctccgaggg ccattgcatg gagccgaaaa gcaaaagcaa cagcgaggca gcatggcgat
9061 ttatcacctt acggcgaaaa ccggcagcag gtcgggcggc caatcggcca gggccaaggc
9121 cgactacatc cagcgcgaag gcaagtatgc ccgcgacatg gatgaagtct tgcacgccga
9181 atccgggcac atgccggagt tcgtcgagcg gcccgccgac tactgggatg ctgccgacct
9241 gtatgaacgc gccaatgggc ggctgttcaa ggaggtcgaa tttgccctgc cggtcgagct
9301 gaccctcgac cagcagaagg cgctggcgtc cgagttcgcc cagcacctga ccggtgccga
9361 gcgcctgccg tatacgctgg ccatccatgc cggtggcggc gagaacccgc actgccacct
9421 gatgatctcc gagcggatca atgacggcat cgagcggccc gccgctcagt ggttcaagcg
9481 gtacaacggc aagaccccgg agaagggcgg ggcacagaag accgaagcgc tcaagcccaa
9541 ggcatggctt gagcagaccc gcgaggcatg ggccgaccat gccaaccggg cattagagcg
9601 ggctggccac gacgcccgca ttgaccacag aacacttgag gcgcagggca tcgagcgcct
9661 gcccggtgtt cacctggggc cgaacgtggt ggagatggaa ggccggggca tccgcaccga
9721 ccgggcagac gtggccctga acatcgacac cgccaacgcc cagatcatcg acttacagga
9781 ataccgggag gcaatagacc atgaacgcaa tcgacagagt gaagaaatcc agaggcatca
9841 acgagttagc ggagcagatc gaaccgctgg cccagagcat ggcgacactg gccgacgaag
9901 cccggcaggt catgagccag acccagcagg ccagcgaggc gcaggcggcg gagtggctga
9961 aagcccagcg ccagacaggg gcggcatggg tggagctggc caaagagttg cgggaggtag
10021 ccgccgaggt gagcagcgcc gcgcagagcg cccggagcgc gtcgcggggg tggcactgga
10081 agctatggct aaccgtgatg ctggcttcca tgatgcctac ggtggtgctg ctgatcgcat
10141 cgttgctctt gctcgacctg acgccactga caaccgagga cggctcgatc tggctgcgct
10201 tggtggcccg atgaagaacg acaggacttt gcaggccata ggccgacagc tcaaggccat
10261 gggctgtgag cgcttcgata tcggcgtcag ggacgccacc accggccaga tgatgaaccg
10321 ggaatggtca gccgccgaag tgctccagaa cacgccatgg ctcaagcgga tgaatgccca
10381 gggcaatgac gtgtatatca ggcccgccga gcaggagcgg catggtctgg tgctggtgga
10441 cgacctcagc gagtttgacc tggatgacat gaaagccgag ggccgggagc ctgccctggt
10501 agtggaaacc agcccgaaga actatcaggc atgggtcaag gtggccgacg ccgcaggcgg
10561 tgaacttcgg gggcagattg cccggacgct ggccagcgag tacgacgccg acccggccag
10621 cgccgacagc cgccactatg gccgcttggc gggcttcacc aaccgcaagg acaagcacac
10681 cacccgcgcc ggttatcagc cgtgggtgct gctgcgtgaa tccaagggca agaccgccac
10741 cgctggcccg gcgctggtgc agcaggctgg ccagcagatc gagcaggccc agcggcagca
10801 ggagaaggcc cgcaggctgg ccagcctcga actgcccgag cggcagctta gccgccaccg
10861 gcgcacggcg ctggacgagt accgcagcga gatggccggg ctggtcaagc gcttcggtga
10921 tgacctcagc aagtgcgact ttatcgccgc gcagaagctg gccagccggg gccgcagtgc
10981 cgaggaaatc ggcaaggcca tggccgaggc cagcccagcg ctggcagagc gcaagcccgg
11041 ccacgaagcg gattacatcg agcgcaccgt cagcaaggtc atgggtctgc ccagcgtcca
11101 gcttgcgcgg gccgagctgg cacgggcacc ggcaccccgc cagcgaggca tggacagggg
11161 cgggccagat ttcagcatgt agtgcttgcg ttggtactca cgcctgttat actatgagta
11221 ctcacgcaca gaagggggtt ttatggaata cgaaaaaagc gcttcagggt cggtctacct
11281 gatcaaaagt gacaagggct attggttgcc cggtggcttt ggttatacgt caaacaaggc
11341 cgaggctggc cgcttttcag tcgctgatat ggccagcctt aaccttgacg gctgcacctt
11401 gtccttgttc cgcgaagaca agcctttcgg ccccggcaag tttctcggtg actgatatga
11461 aagaccaaaa ggacaagcag accggcgacc tgctggccag ccctgacgct gtacgccaag
11521 cgcgatatgc cgagcgcatg aaggccaaag ggatgcgtca gcgcaagttc tggctgaccg
11581 acgacgaata cgaggcgctg cgcgagtgcc tggaagaact cagagcggcg cagggcgggg
11641 gtagtgaccc cgccagcgcc taaccaccaa ctgcctgcaa aggaggcaat caatggctac
11701 ccataagcct atcaatattc tggaggcgtt cgcagcagcg ccgccaccgc tggactacgt
11761 tttgcccaac atggtggccg gtacggtcgg ggcgctggtg tcgcccggtg gtgccggtaa
11821 atccatgctg gccctgcaac tggccgcaca gattgcaggc gggccggatc tgctggaggt
11881 gggcgaactg cccaccggcc cggtgatcta cctgcccgcc gaagacccgc ccaccgccat
11941 tcatcaccgc ctgcacgccc ttggggcgca cctcagcgcc gaggaacggc aagccgtggc
12001 tgacggcctg ctgatccagc cgctgatcgg cagcctgccc aacatcatgg ccccggagtg
12061 gttcgacggc ctcaagcgcg ccgccgaggg ccgccgcctg atggtgctgg acacgctgcg
12121 ccggttccac atcgaggaag aaaacgccag cggccccatg gcccaggtca tcggtcgcat
12181 ggaggccatc gccgccgata ccgggtgctc tatcgtgttc ctgcaccatg ccagcaaggg
12241 cgcggccatg atgggcgcag gcgaccagca gcaggccagc cggggcagct cggtactggt
12301 cgataacatc cgctggcagt cctacctgtc gagcatgacc agcgccgagg ccgaggaatg
12361 gggtgtggac gacgaccagc gccggttctt cgtccgcttc ggtgtgagca aggccaacta
12421 tggcgcaccg ttcgctgatc ggtggttcag gcggcatgac ggcggggtgc tcaagcccgc
12481 cgtgctggag aggcagcgca agagcaaggg ggtgccccgt ggtgaagcct aagaacaagc
12541 acagcctcag ccacgtccgg cacgacccgg cgcactgtct ggcccccggc ctgttccgtg
12601 ccctcaagcg gggcgagcgc aagcgcagca agctggacgt gacgtatgac tacggcgacg
12661 gcaagcggat cgagttcagc ggcccggagc cgctgggcgc tgatgatctg cgcatcctgc
12721 aagggctggt ggccatggct gggcctaatg gcctagtgct tggcccggaa cccaagaccg
12781 aaggcggacg gcagctccgg ctgttcctgg aacccaagtg ggaggccgtc accgctgatg
12841 ccatggtggt caaaggtagc tatcgggcgc tggcaaagga aatcggggca gaggtcgata
12901 gtggtggggc gctcaagcac atacaggact gcatcgagcg cctttggaag gtatccatca
12961 tcgcccagaa tggccgcaag cggcaggggt ttcggctgct gtcggagtac gccagcgacg
13021 aggcggacgg gcgcctgtac gtggccctga accccttgat cgcgcaggcc gtcatgggtg
13081 gcggccagca tgtgcgcatc agcatggacg aggtgcgggc gctggacagc gaaaccgccc
13141 gcctgctgca ccagcggctg tgtggctgga tcgaccccgg caaaaccggc aaggcttcca
13201 tagatacctt gtgcggctat gtctggccgt cagaggccag tggttcgacc atgcgcaagc
13261 gccgccagcg ggtgcgcgag gcgttgccgg agctggtcgc gctgggctgg acggtaaccg
13321 agttcgcggc gggcaagtac gacatcaccc ggcccaaggc ggcaggctga ccccccccac
13381 tctattgtaa acaagacatt tttatctttt atattcaatg gcttattttc ctgctaattg
13441 gtaataccat gaaaaatacc atgctcagaa aaggcttaac aatattttga aaaattgcct
13501 actgagcgct gccgcacagc tccataggcc gctttcctgg ctttgcttcc agatgtatgc
13561 tcttctgctc ccgaacgcca gcaagacgta gcccagcgcg tcggccagct tgcaattcgc
13621 gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
13681 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgcc
13741 agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt caccgcctgg
13801 ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt
13861 ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc gtatcccact
13921 accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc
13981 gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt cagcatttgc
14041 atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc tatcggctga
14101 atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc cgagacagaa
14161 cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag atgctccacg
14221 cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt ctggtcagag
14281 acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat ggcatcctgg
14341 tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc
14401 gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac gctggcaccc
14461 agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg cagggccaga
14521 ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg
14581 ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa
14641 acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc ggcatactct
14701 gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact ctcttccggg
14761 cgctatcatg ccataccgcg aaaggttttg caccattcga tggtgtcaac gtaaatgccg
14821 cttcgccttc gcgcgcgaat tgcaagctga tccgggctta tcgactgcac ggtgcaccaa
14881 tgcttctggc gtcaggcagc catcggaagc tgtggtatgg ctgtgcaggt cgtaaatcac
14941 tgcataattc gtgtcgctca aggcgcactc ccgttctgga taatgttttt tgcgccgaca
15001 tcataacggt tctggcaaat attctgaaat gagctgttga caattaatca tcggctcgta
15061 taatgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagaatt cgagctcggt
15121 accaggagga aacgatgaat gcgccagtaa atggaacact tattactaca cagattgcaa
15181 acgttgctga aactcttggc ttggttaatg ttaatccaca agagttaaag gaaacactga
15241 ttcaaacagc tttccgtact gaaacacctg caactgatgc acaaatggct tctcttttga
15301 ttgttgctgg tcaatacaag ctgaacccat ggaccaaaga gatttacgct ttcccagata
15361 aaaacaaagg gattattcca gttgttggcg tggatggttg gtctcgaatc attaatggaa
15421 actctaattt caatggtatg gaatttaagt tttctgaaaa tatggttcag atggaaggcg
15481 cgaaagttgc tgcacctgaa tgggttgaat gcattatcta tcgtaaagat cgtgaccacc
15541 ctactgttgt tcgcgagtat ttagcagagt gttatcgtgc accatttaag tcaaaaactg
15601 gatatgtagt tgaaggacca tggcagagtc acccttctcg cttcttgcgc cacaaagcaa
15661 ctattcagtg tgcacgctta gcctttggtt ttgtgggtat tcatgatcag gatgaagcag
15721 aacgtatcgc tgaaagtgga caagctatta aggatgtgac tagtgaagtg ccagaaggct
15781 accaagcctt tgaagatgag catttgccta cactcaaatc agaagctcaa tacggcactg
15841 aacgcttgca agctgcttat gtggcaattc caaagggaaa tcttaaaaag cacctttggg
15901 aagttcactc aattagctta aaaaaaattg ctcagtttgc agaccaagct ttacagcgcc
15961 aaggagaaac atatgaacat tctccagcgt agtgaagatt ggcattcgga acgctgtggg
16021 aaagtcacag caagccgtgt aaaggattta aatgcaaagc tcaataaagg aaaagcttta
16081 aatgcattgg gtttaactat tctagctgag cgcctcactg gcgttcagaa ggaaatcttc
16141 actaaccaag ctatgcaatg gggtatcgat aacgagcctc atgcaatagc agcttatgaa
16201 aatgaaacgg gtaactttgt agtaggtaca ggtttaattg accatccttt cattgaaatg
16261 ttcggggctt caccagatgg gcttgtgggc gacaatgggc aaatagaagt taagtgccca
16321 gatactacaa cacacttgaa caccctgctg accaagcaag taccagatga gtacattcct
16381 caaataacta gtcagttggc ttgtactcgt cgtgaatggt gtgactttgt gagttatgac
16441 ccacgtctac cagaaggatt acagatcatc attattcgtg tctatgccaa agacctagca
16501 atagaagcac ttgagcaaga tgttcgtaag ttcaacaaag ctatagatga cgcaattaaa
16561 acattgaagg tggcagcatg agtcgacctg caggcatgca
//
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