| 质粒名称: | pKCcas9dO |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 62552 |
| 存储实验室: | Weihong Jiang |
|
载体骨架基本信息 |
|
| 载体名称: | pKC1139 |
| 载体出品公司: | -- |
| 载体抗性: | 低安普霉素(25 ug/mL) |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，CRISPR；细菌基因组编辑 |
| 筛选标记: | -- |
| 高拷贝/低拷贝: | 低拷贝 |
| 生长条件: | 30 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | Scocas9 |
| 物种来源: | artificially synthesized, codon optimized gene from Strep pyogenes |
| 插入大小: | 4107 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | ptipA |
| 标签/融合蛋白: | -- |
| 克隆方法: | 限制性内切酶 |
| 5' 克隆位点: | NdeI(未破坏) |
| 3' 克隆位点: | EcoRI（销毁后未知） |
| 5' 测序引物 | -- |
| 3' 测序引物 | M13pUC-Rev |
|
文献引用方法 |
|
| 材料和方法部分: | pKCcas9dO was a gift from Weihong Jiang (Addgene plasmid # 62552 ; http://n2t.net/addgene:62552 ; RRID:Addgene_62552) |
| 参考文献部分: | One-step high-efficiency CRISPR/Cas9-mediated genome editing in Streptomyces. Huang H, Zheng G, Jiang W, Hu H, Lu Y. Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):231-43. doi: 10.1093/abbs/gmv007. Epub 2015 Mar 3. 10.1093/abbs/gmv007 PubMed 25739462 |
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 pKCcas9dO 13375 bp ds-DNA circular SYN 05-MAR-2019
DEFINITION High efficiency Streptomyces genome editing by CRISPR/Cas9 system.
ACCESSION .
VERSION .
KEYWORDS pKCcas9dO
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13375)
AUTHORS Huang H, Zheng G, Jiang W, Hu H, Lu Y
TITLE One-step high-efficiency CRISPR/Cas9-mediated genome editing in
Streptomyces.
JOURNAL Acta Biochim Biophys Sin (Shanghai). 2015 Apr;47(4):231-43. doi:
10.1093/abbs/gmv007. Epub 2015 Mar 3.
PUBMED 25739462
REFERENCE 2 (bases 1 to 13375)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Mar 5, 2019 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..13375
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(547..1989)
/codon_start=1
/gene="rep"
/product="replication initiator protein from the
Streptomyces ghanaensis plasmid pSG5"
/label=pSG5 Rep
/translation="MWKPGEATWGNTCRCNNVHTCPWCMSRILAVRGSNVQLAADGLAD
AGYGLHLGTNTLRHFERMAFGTVRKGMRHGLVAVLHDGWKGAYGSSGRRWRTMRDDFGI
IGYERAFEDTFGWGSGWHLHWHTLWVTREVLGPDAQAAFRDALAGAWAAGVESAGGYTV
SETCDRPGCSCEGKGHGTDVRPLNGADAADGDAGKQARYLYKDGDKTKGGVAKIGLELA
GQNFKAGRGDDRMGPLDLGDAAAAELQRLRRPGPFVEKYREREFGVFQVRKHYRSQNLN
RLIKELGIQQDVRTEEEITDDTEGLVAIAVIPAYIWYRYIARVAGRRLDLIKVAETYGL
PGVRRLVESWGLVWGKDVLDPPAPEAPAAPGDLDADQMRFEVMSEEEAAFREARRKANE
ARTEELAASLDRVRQPKKEAIRPTISLRKRLKPKPVTVDVKTPPPGAASPVCRRCKGKL
APVLQPWGQHPGDCLRVDTAVA"
oriT 3156..3265
/note="incP origin of transfer"
CDS 3298..3669
/codon_start=1
/gene="traJ"
/product="oriT-recognizing protein"
/label=traJ
/translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
KQDELGKVMMGVVRPRAEP"
primer_bind 4355..4377
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 4369..4386
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 4370..4386
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
misc_RNA complement(6428..6503)
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
promoter complement(6524..6558)
/label=J23119(SpeI) promoter
/note="bacterial promoter (Registry of Standard Biological
Parts BBa_J23119) modified to end with an SpeI site"
CDS 6770..10876
/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"
primer_bind complement(10891..10907)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10891..10907)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10904..10926)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 10915..10931
/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(10939..10969)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 10984..11005
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11122..11139)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(11293..11881)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(11373..11392)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 12085..12885
/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"
ORIGIN
1 tgtgaggagg ctggtccttt accctgatgc ccgagggcat cagggtaaag gaccagcctc
61 gccaactagg agcgctacca aggcgaacac aacccaggac ttggtagaac ccttcgccgg
121 agaagttcag actcatcggc atgagcaccc ctcgccgcgc actcggcacc ggtccgtcga
181 ccaccaccag gacgtcgttg tcgacgtcgg ccccgcggct cctgcccgcc gaacgcgtcg
241 tcgtcgacgg cctggtgctc atcgacgagc acccggagcc aggtgaaaag cgccggcgga
301 cgctcggact gggcgcggga ttccagcagt aacccaggtc cgccggccac ctcacggcag
361 gcagaccctc ggctttcgcg ccgggaccgc catgagaccg ccacccggat gtccggggtg
421 gcggtctcat ggcggtccct cagcggccct acgcgaccgc tgtgtcgacg cggaggcagt
481 ctccggggtg ctggccccag ggcyggakkw ccggggyggc ggtctcatgg cggtccctca
541 gcggccctac gcgaccgctg tgtcgacgcg gaggcagtct ccggggtgct ggccccaggg
601 ctggagtacc ggggcgagct tgcccttgca gcggcggcac acgggcgacg cggcgccggg
661 cgggggagtc ttgacgtcga cggtgaccgg cttcggcttc aggcgcttcc gaaggctgat
721 cgtcgggcgg atcgcctcct tcttcggctg acgcactcgg tcgagcgaag cggccagctc
781 ctccgtgcgg gcctcgttgg ccttccggcg cgcctcgcgg aaagcagctt cctcctcgga
841 catgacctcg aacctcatct ggtcagcgtc aaggtcgccc ggcgcggccg gcgcttccgg
901 ggcgggcggg tccaggacgt ccttgcccca caccaggccc caggactcga cgagccgccg
961 gacgcccggt aggccgtacg tctcggcgac cttgatgagg tcgaggcgac gtccggcgac
1021 gcgggcgatg tatcggtacc agatgtaggc cgggatgacc gcgatggcga ccaggccctc
1081 ggtgtcgtcg gtgatctcct cctcggtgcg gacgtcctgc tggatgccga gttccttgat
1141 cagccggttc aggttctgcg accggtagtg cttgcggacc tggaagacgc cgaactcgcg
1201 ctcgcggtac ttctcgacga acgggccggg ccgacgaagc cgctgcagct cggcggccgc
1261 cgcgtcgccc aggtcgagcg gtcccatgcg gtcgtcgccg cgaccggcct tgaagttctg
1321 tccggccagc tccaggccga tcttggcgac gccgcccttg gtcttgtcgc cgtccttgta
1381 gaggtagcgg gcctgcttgc ccgcatcgcc gtcagcggcg tccgcgccgt tgagtgggcg
1441 cacgtcggtg ccgtggccct tgccctcaca ggagcaaccg ggccggtcgc acgtctcgct
1501 gacggtgtag ccgcccgcgg attcgacccc ggcggcccag gctccggcga gtgcgtcgcg
1561 gaacgcggcc tgggcgtccg ggccgagcac ctcgcgggtg acccagagcg tgtgccagtg
1621 caggtgccag ccggagcccc agccgaaggt gtcctcgaag gcccgctcgt agccgatgat
1681 cccgaagtcg tcgcgcatcg tgcgccagcg gcggccggac gagccgtacg cgcccttcca
1741 gccgtcgtgc aagaccgcga ccaggccgtg ccgcattccc ttgcggacgg tgccgaacgc
1801 catgcgctcg aagtggcgca acgtgttcgt gccaaggtgc agcccgtacc cggcgtccgc
1861 gagaccgtcg gcggcgagct gcacgttcga gccccgtacg gccaggatgc ggctcatgca
1921 ccacgggcag gtgtggacgt tgttgcagcg gcacgtgttg ccccacgtcg cctcgcccgg
1981 cttccacatc agctcggccg tcccggcagt gagccgggtc ccgcagccct tgaacgcctc
2041 gttcagcgac accgtctggt gccggtcccg ccgggcgaac cgctcgtcgc gcgggtcctc
2101 ccgcccggct gtcgcggcac cctcgtttgg ggtagaaccc gttccagtta cagcgctctg
2161 acctgcagtg gacggagatt ttcccttact actaaagccc gcgtccggat tacccgctgt
2221 agtcgtgctt gctacgctgc gtgactggtc cgcaatgaga cgctttgcgc gctttcggca
2281 ggcgtccgag cagtagattt tggggcgctt cccggggatg tggacgatcg gggtgccgca
2341 gtggcacttc ggtccggcgg ggcgcggtgg tgtcgacgcg ctgttctctc gtacgctcgt
2401 cacagagcaa acgtcctcac tcggcatgct gcgccggttc gggggcggcg agcccgggag
2461 gccaatcccg ggctcgtgcc atttctgggt cctgttgatc ctggcattgg tgtggccgtt
2521 cattgcccct gctcgctcct gacgcgccga tagacgtccg atacgcccgg tgctggtggg
2581 atttgatagg tcggaagaag ccccgcccgg gcctgggcgg ggcttcctgt gcgtcaggac
2641 ctcctcgtcg tgagcctctt cggcctatgg acggagtgac ctcgtgatcc gttacagccg
2701 cgcgcgctcg cgtagagcgg tctcatcagt tccacgaacg gtcctcttcg cagatcaggg
2761 cgttggggcg gagtctcacc aaggactacg tctgctggcg atttccgtta caccccgggc
2821 ggtggccggc gcacacgcgc gcccgcgttg ggcagtgcag aaagtgcaga aacctaggcg
2881 ctgatggtcc aggtccacgg ttcgtcgtcg gcggcggcgc gggcggcggc gtcggccagg
2941 gcgcgggcga gaccggctac ggcgggcttg atgcgccggt tgcgggcgac cttgagcagc
3001 tagtatgcag gtcgacggat cttttccgct gcataaccct gcttcggggt cattatagcg
3061 attttttcgg tatatccatc ctttttcgca cgatatacag gattttgcca aagggttcgt
3121 gtagactttc cttggtgtat ccaacggcgt cagccgggca ggataggtga agtaggccca
3181 cccgcgagcg ggtgttcctt cttcactgtc ccttattcgc acctggcggt gctcaacggg
3241 aatcctgctc tgcgaggctg gccggctacc gccggcgtaa cagatgaggg caagcggatg
3301 gctgatgaaa ccaagccaac caggaagggc agcccaccta tcaaggtgta ctgccttcca
3361 gacgaacgaa gagcgattga ggaaaaggcg gcggcggccg gcatgagcct gtcggcctac
3421 ctgctggccg tcggccaggg ctacaaaatc acgggcgtcg tggactatga gcacgtccgc
3481 gagctggccc gcatcaatgg cgacctgggc cgcctgggcg gcctgctgaa actctggctc
3541 accgacgacc cgcgcacggc gcggttcggt gatgccacga tcctcgccct gctggcgaag
3601 atcgaagaga agcaggacga gcttggcaag gtcatgatgg gcgtggtccg cccgagggca
3661 gagccatgac ttttttagcc gctaaaacgg ccggggggtg cgcgtgattg ccaagcacgt
3721 ccccatgcgc tccatcaaga agagcgactt cgcggagctg gtgaagtaca tcaccgacga
3781 gcaaggcaag accgatcccc ggggacctgc atagatctac gtctgtcgag aagtttctga
3841 tygmmaagyw cgwcmscrts cgctccatca agaagagcga cttcgcggag ctggtgaagt
3901 acatcaccga cgagcaaggc aagaccgatc cccggggacc tgcatagatc tacgtctgtc
3961 gagaagtttc tgatcgaaaa gttcgacagc gtctccgacc tgatgcagct ctcggagggc
4021 gaagaatctc gtgctttcag cttcgatgta ggagggcgtg gatatgtcct gcgggtaaat
4081 agctgcgccg atggtttcta caaagatcgt tatgtttatc ggcactttgc atcggccgcg
4141 ctcccgattc cggaagtgct tgacattggg gaatttatgc ggtgtgaaat accgcacaga
4201 tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg
4261 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
4321 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
4381 gccagtgcca agcttcgacc gccgctcgag ccgggcgccc aaagctgagc gcgtgaggga
4441 ccacgaggca gggaggggaa cacatggcca cgcaggactc cgaagtcgca ctggtgacgg
4501 gtgcgaccag cggaatcggg ctggagatcg cccgcaggct cggtaaggag gggttgcgcg
4561 tgttcgtctg cgcgcgcggc gaagaaggac tgcggacgac gctgaaggag ttgcgggagg
4621 cgggcgtgga ggcggacggg cggacctgcg acgtccgctc ggtcccggag atcgaggctc
4681 tcgtggcggc ggtggtcgag cgttacggtc cggtcgacgt gctggtgaac aacgcgggcc
4741 ggcccggcgg cggcgccacg gccgaactcg ccgacgaact ctggctcgat gtcgtggaga
4801 ccaacctcac cggcgtgttc cgggtgacca aacaggtcct caaggcgggc ggcatgctcg
4861 aacggggcac gggccgaatc gtcaacatcg cctcgaccgg cggaaagcag ggtgtggtgc
4921 acgccgcgcc ctactccgcc tcgaagcacg gcgtggtcgg cttcaccaag gcgctcggtc
4981 tcgaactggc gaggaccggc atcacggtga acgccgtctg ccccggattc gtcgagacgc
5041 cgatggccgc gtccgtgcgc gagcactact cggacatctg ggaggtgtcg accgaggagg
5101 ccttcgaccg gatcaccgcg cgcgtgccga tcggccggta cgtgcagccg tccgaggtgg
5161 cggagatggt ggcgtacctg atcgggcccg gtgcggccgc gttcacggac cggtgctgcg
5221 cccccgtcgr grtycwycgk cysstrsgkg crgccgtccg aggtggcgga gatggtggcg
5281 tacctgatcg ggcccggtgc ggccgcgttc acggaccggt gctgcgcccc cgtcgagatt
5341 ctccgtctcc tgggggcgac aataacaggc ctactcaaca gatttcaatg tgcttgccgt
5401 cacaagcgat ctcctattga tcaaaaatta atcagtgcag cgagctgcac tgattaattt
5461 ttgctggatt ttaccgagag gcagtggtcg aggcttccga atcaatagaa tctggcattc
5521 ctgatacggc tgggtcctcg gaagggtcct cgcgccgggc gagcgggtgc ggccaccaca
5581 tccaccggcc gacatcgaga gtgagcgcgg tgaccaggac cgatcgcacg atcatggtgt
5641 cgagcaggac gcccaccgcg accgtgaatc cgagctcggc gatgaagacc agcggcagcg
5701 tggccagcgc ggcgaaggtc ccggccagca ccagcccggc cgcggtgatc acgccaccgg
5761 tcgaggtgag gcccttgagc gcgccctgcc gggtgccgag ccggtcgctc tcctccttga
5821 tccgggtgac gaggaagatg ttgtagtcga cgcccagggc gaccaggaag acgaagaccc
5881 acagggggaa ggccgaatcg gctcccgcga agtcgaagac gtggttgaag aagagggtgg
5941 cgaggccgag cgcggtgaag aaggacagca ccacgctcgc gatcagcagc agcggcgcca
6001 ccagtgctcg cagcagcagc gcgaggatgc agaacaccac ggcgaggatc accggaatga
6061 tcagaccgcg gtcgcggctg gacgcctccc gcatgtcgtg cacgacggcg ctgctgccgc
6121 ccacccgtgc ctgcgcaccg tccagccggg cgagtgtctc gcgggccgcg gtgaccgaac
6181 gcatggccgc cggcgagtcc gccccggcgc cgagcgtcgc ctcgacatag gcgaggccgt
6241 ccttcacctg tggtggtgcg acctcgatga cgccgggcac ggtctccaga gctgcgtgca
6301 cctggtccgc cgaggcgccc ttggcgatga cgaccatggg ttcaccggag ccggccggga
6361 agtgccgggt caggatctcc tcgcccacca ccgactccgg cttggtggaa ttcagatctc
6421 aaaaaaagca ccgactcggt gccacttttt caagttgata acggactagc cttattttaa
6481 cttgctattt ctagctctaa aacgaactcc ggtccctggt aatactagta ttatacctag
6541 gactgagcta gctgtcaagg atccgcggcc gcgcgcgatc tagagcaacc gagcgttctg
6601 aacaaatcca gatggagttc tgaggtcatt actggaccgg atcggggatc tgggctgagg
6661 gagccgacgg cacgcggcgg ctcacggcgt ggcacgcgga acgtccgggc ttgcacctca
6721 cgtcacgtga ggaggcagcg tggacggcgt cagagaaggg agcggacata tggacaagaa
6781 gtactccatc ggcctggaca tcggcaccaa ctccgtcggc tgggccgtca tcacggacga
6841 gtacaaggtc ccgtcgaaga agttcaaggt gctgggcaac acggaccgcc acagcatcaa
6901 gaagaacctc atcggcgccc tcctgttcga cagcggcgag acggccgagg cgacccgcct
6961 gaagcgcacg gcgcggcggc ggtacacgcg ccgcaagaac cgcatctgct acctccagga
7021 gatcttctcc aacgagatgg cgaaggtcga cgactcgttc ttccaccggc tggaggagtc
7081 cttcctggtc gaggaggaca agaagcacga gcggcacccc atcttcggca acatcgtgga
7141 cgaggtggcc taccacgaga agtacccgac catctaccac ctccggaaga agctggtgga
7201 ctcgaccgac aaggccgacc tgcgcctgat ctacctggcc ctcgcccaca tgatcaagtt
7261 ccgcggccac ttcctcatcg agggcgacct gaacccggac aactcggacg tcgacaagct
7321 gttcatccag ctcgtgcaga cctacaacca gctcttcgag gagaacccga tcaacgcctc
7381 cggcgtggac gcgaaggcca tcctgtcggc ccgcctctcg aagtcccggc ggctcgagaa
7441 cctgatcgcc cagctgccgg gcgagaagaa gaacggcctc ttcggcaacc tgatcgcgct
7501 ctccctgggc ctgacgccga acttcaagtc caacttcgac ctcgccgagg acgccaagct
7561 ccagctgtcc aaggacacgt acgacgacga cctggacaac ctcctggcgc agatcggcga
7621 ccagtacgcg gacctgttcc tcgcggcgaa gaacctctcc gacgcgatcc tgctcagcga
7681 catcctccgc gtcaacaccg agatcaccaa ggcgcccctg tccgcgagca tgatcaagcg
7741 ctacgacgag caccaccagg acctcaccct gctgaaggcc ctggtccgcc agcagctccc
7801 ggagaagtac aaggagatct tcttcgacca gtccaagaac ggctacgcgg gctacatcga
7861 cggcggcgcg tcccaggagg agttctacaa gttcatcaag cccatcctgg agaagatgga
7921 cggcaccgag gagctgctgg tcaagctgaa ccgggaggac ctcctgcgca agcagcgcac
7981 gttcgacaac ggctccatcc cccaccagat ccacctgggc gagctgcacg ccatcctccg
8041 gcgccaggag gacttctacc cgttcctcaa ggacaaccgc gagaagatcg agaagatcct
8101 cacgttccgg atcccgtact acgtcggccc cctcgcccgc ggcaactccc gcttcgcgtg
8161 gatgacgcgc aagtcggagg agacgatcac cccgtggaac ttcgaggagg tggtggacaa
8221 gggcgcctcc gcgcagtcgt tcatcgagcg gatgaccaac ttcgacaaga acctgccgaa
8281 cgagaaggtg ctgcccaagc acagcctcct gtacgagtac ttcacggtct acaacgagct
8341 caccaaggtg aagtacgtga cggagggcat gcggaagccg gcgttcctgt ccggcgagca
8401 gaagaaggcc atcgtggacc tgctgttcaa gacgaaccgg aaggtcaccg tcaagcagct
8461 gaaggaggac tacttcaaga agatcgagtg cttcgactcg gtcgagatca gcggcgtgga
8521 ggaccgcttc aacgcgtcgc tgggcacgta ccacgacctc ctgaagatca tcaaggacaa
8581 ggacttcctc gacaacgagg agaacgagga catcctggag gacatcgtgc tgaccctgac
8641 gctcttcgag gaccgggaga tgatcgagga gcgcctcaag acctacgcgc acctgttcga
8701 cgacaaggtc atgaagcagc tcaagcggcg gcggtacacc ggctggggcc ggctctcccg
8761 caagctcatc aacggcatcc gggacaagca gtcgggcaag accatcctcg acttcctgaa
8821 gtccgacggc ttcgccaacc gcaacttcat gcagctgatc cacgacgaca gcctgacgtt
8881 caaggaggac atccagaagg cgcaggtgtc gggccagggc gactccctgc acgagcacat
8941 cgcgaacctg gccggcagcc cggccatcaa gaagggcatc ctgcagacgg tcaaggtggt
9001 cgacgagctg gtgaaggtga tgggccgcca caagcccgag aacatcgtca tcgagatggc
9061 ccgggagaac cagaccacgc agaagggcca gaagaacagc cgcgagcgca tgaagcggat
9121 cgaggagggc atcaaggagc tcggctcgca gatcctcaag gagcaccccg tcgagaacac
9181 ccagctccag aacgagaagc tctacctcta ctacctgcag aacggccggg acatgtacgt
9241 ggaccaggag ctcgacatca accggctgtc ggactacgac gtcgaccaca tcgtcccgca
9301 gtcgttcctg aaggacgaca gcatcgacaa caaggtcctc acccgctcgg acaagaaccg
9361 gggcaagtcc gacaacgtcc cgtcggagga ggtcgtgaag aagatgaaga actactggcg
9421 gcagctcctc aacgccaagc tcatcacgca gcgcaagttc gacaacctga cgaaggcgga
9481 gcggggcggc ctcagcgagc tcgacaaggc gggcttcatc aagcggcagc tggtcgagac
9541 gcggcagatc acgaagcacg tggcccagat cctggactcg cgcatgaaca ccaagtacga
9601 cgagaacgac aagctgatcc gcgaggtcaa ggtcatcacc ctcaagtcca agctggtgag
9661 cgacttccgc aaggacttcc agttctacaa ggtgcgggag atcaacaact accaccacgc
9721 ccacgacgcg tacctgaacg cggtggtggg cacggcgctg atcaagaagt acccgaagct
9781 cgagagcgag ttcgtctacg gcgactacaa ggtgtacgac gtgcggaaga tgatcgcgaa
9841 gagcgagcag gagatcggca aggccacggc gaagtacttc ttctacagca acatcatgaa
9901 cttcttcaag accgagatca ccctggcgaa cggcgagatc cgcaagcgcc cgctcatcga
9961 gaccaacggc gagacgggcg agatcgtgtg ggacaagggc cgcgacttcg cgaccgtccg
10021 gaaggtgctg tccatgcccc aggtcaacat cgtcaagaag acggaggtcc agaccggcgg
10081 cttcagcaag gagtccatcc tgcccaagcg gaacagcgac aagctgatcg cccggaagaa
10141 ggactgggac cccaagaagt acggcggctt cgactccccc accgtcgcct actcggtcct
10201 cgtggtcgcc aaggtggaga agggcaagag caagaagctg aagtccgtga aggagctgct
10261 gggcatcacc atcatggagc ggtcgtcgtt cgagaagaac cccatcgact tcctggaggc
10321 caagggctac aaggaggtga agaaggacct catcatcaag ctccccaagt actccctgtt
10381 cgagctggag aacggccgga agcgcatgct cgccagcgcg ggcgagctgc agaagggcaa
10441 cgagctggcg ctgccgagca agtacgtcaa cttcctgtac ctcgcctcgc actacgagaa
10501 gctgaagggc tcgccggagg acaacgagca gaagcagctg ttcgtggagc agcacaagca
10561 ctacctcgac gagatcatcg agcagatctc cgagttctcc aagcgcgtca tcctggccga
10621 cgcgaacctg gacaaggtcc tgtccgcgta caacaagcac cgcgacaagc cgatccggga
10681 gcaggccgag aacatcatcc acctgttcac gctgaccaac ctcggcgccc ccgcggcctt
10741 caagtacttc gacaccacga tcgaccggaa gcggtacacg agcacgaagg aggtgctcga
10801 cgccaccctg atccaccaga gcatcacggg cctgtacgag acccgcatcg acctctccca
10861 gctgggcggc gactgagaat tcgtaatcat gtcatagctg tttcctgtgt gaaattgtta
10921 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
10981 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
11041 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
11101 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
11161 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
11221 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
11281 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
11341 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
11401 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
11461 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
11521 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
11581 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
11641 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
11701 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
11761 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
11821 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
11881 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
11941 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt tggttcatgt
12001 gcagctccat cagcaaaagg ggatgataag tttatcacca ccgactattt gcaacagtgc
12061 cgttgatcgt gctatgatcg actgatgtca tcagcggtgg agtgcaatgt cgtgcaatac
12121 gaatggcgaa aagccgagct catcggtcag cttctcaacc ttggggttac ccccggcggt
12181 gtgctgctgg tccacagctc cttccgtagc gtccggcccc tcgaagatgg gccacttgga
12241 ctgatcgagg ccctgcgtgc tgcgctgggt ccgggaggga cgctcgtcat gccctcgtgg
12301 tcaggtctgg acgacgagcc gttcgatcct gccacgtcgc ccgttacacc ggaccttgga
12361 gttgtctctg acacattctg gcgcctgcca aatgtaaagc gcagcgccca tccatttgcc
12421 tttgcggcag cggggccaca ggcagagcag atcatctctg atccattgcc cctgccacct
12481 cactcgcctg caagcccggt cgcccgtgtc catgaactcg atgggcaggt acttctcctc
12541 ggcgtgggac acgatgccaa cacgacgctg catcttgccg agttgatggc aaaggttccc
12601 tatggggtgc cgagacactg caccattctt caggatggca agttggtacg cgtcgattat
12661 ctcgagaatg accactgctg tgagcgcttt gccttggcgg acaggtggct caaggagaag
12721 agccttcaga aggaaggtcc agtcggtcat gcctttgctc ggttgatccg ctcccgcgac
12781 attgtggcga cagccctggg tcaactgggc cgagatccgt tgatcttcct gcatccgcca
12841 gaggcgggat gcgaagaatg cgatgccgct cgccagtcga ttggctgagc tcatgagcgg
12901 agaacgagat gacgttggag gggcaaggtc gcgctgattg ctggggcaac acgtggagcg
12961 gatcggggat tgtctttctt cagctcgctg atgatatgct gacgctcaat gccgtttggc
13021 ctccgactaa cgaaaatccc gcatttggac ggctgatccg attggcacgg cggacggcga
13081 atggcggagc agacgctcgt ccgggggcaa tgagatatga aaaagcctga actcaccgcg
13141 acgtatcgat gtcgaggttc ctcaggggag ccaccccaga gaagccctcg gagctgagcg
13201 gagctatttc caaagccatg ccagctagag acagtgcaca ccgccaagca tctgcaaaac
13261 cctcgcctgg agggaaagtg caatgtacgt acgctggttt ccctccagaa agggattctg
13321 cggcttctca actttggaag aagaggcgga acgaactgct gtttcagcct actca
//
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