| 质粒名称: | pCRISPomyces-2 |
|---|---|
| 出品公司: | Addgene |
| 目录编号: | 61737 |
| 存储实验室: | Huimin Zhao |
|
载体骨架基本信息 |
|
| 载体名称: | pKC1139 |
| 载体出品公司: | -- |
| 载体抗性: | 安普霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 10995 bp |
| 载体修饰: | 突变G8694C从pSG5-rep中去除BbsI位点 |
| 载体类型: | 细菌表达，CRISPR |
| 筛选标记: | -- |
| 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: | sSpCas9,Streptomyces codon-optimized Cas9 |
| 物种来源: | Synthetic |
| 插入大小: | 4107 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | rpsL(XC)-BbsI |
| 标签/融合蛋白: | -- |
| 克隆方法: | -- |
| 5' 克隆位点: | -- |
| 3' 克隆位点: | -- |
| 5' 测序引物 | -- |
| 3' 测序引物 | -- |
|
文献引用方法 |
|
| 材料和方法部分: | pCRISPomyces-2 was a gift from Huimin Zhao (Addgene plasmid # 61737 ; http://n2t.net/addgene:61737 ; RRID:Addgene_61737) |
| 参考文献部分: | High-Efficiency Multiplex Genome Editing of Streptomyces Species Using an Engineered CRISPR/Cas System. Cobb RE, Wang Y, Zhao H. ACS Synth Biol. 2014 Dec 8. 10.1021/sb500351f PubMed 25458909 |
买家导航
pCRISPomyces-2载体 pCRISPomyces2质粒基本信息
pCRISPomyces-2质粒图谱 pCRISPomyces2载体图谱和pCRISPomyces-2载体序列 pCRISPomyces2质粒序列多克隆位点信息
pCRISPomyces-2质粒 pCRISPomyces2载体简介
pCRISPomyces-2质粒序列 pCRISPomyces2载体序列
LOCUS pCRISPomyces-2 10994 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Streptomyces expression of codon-optimized Cas9 and custom gRNA.
ACCESSION .
VERSION .
KEYWORDS pCRISPomyces-2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10994)
AUTHORS Cobb RE, Wang Y, Zhao H
TITLE High-Efficiency Multiplex Genome Editing of Streptomyces Species
Using an Engineered CRISPR/Cas System.
JOURNAL ACS Synth Biol. 2014 Dec 8.
PUBMED 25458909
REFERENCE 2 (bases 1 to 10994)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..10994
/organism="synthetic DNA construct"
/mol_type="other DNA"
CDS complement(153..1595)
/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 2762..2871
/note="incP origin of transfer"
promoter 3093..3394
/label=rpsL promoter
/note="promoter for the Xylanimonas cellulosilytica
ribosomal protein S12 gene"
CDS 3395..7501
/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"
promoter 7701..7985
/label=gapdh promoter
/note="glyceraldehyde-3-phosphate dehydrogenase promoter
from Eggerthella lenta"
CDS complement(7994..8290)
/codon_start=1
/gene="lacZ fragment"
/product="LacZ-alpha fragment of beta-galactosidase"
/label=lacZ-alpha
/translation="MTMITPSAQLTLTKGNKSWRTSRGGPVPNSPYSESYYARSLAVVL
QRRDWENPGVTQLNRLAAHPPFASWRNSEEARTDRPSQQLRSLNGEWDAPCSG"
primer_bind 8140..8162
/label=M13/pUC Forward
/note="In lacZ gene"
primer_bind 8154..8171
/label=M13 Forward
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind 8155..8171
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 8181..8200
/label=T7
/note="T7 promoter, forward primer"
promoter 8181..8199
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(8247..8267)
/label=T3
/note="T3 promoter, forward primer"
promoter complement(8247..8265)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(8286..8302)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(8286..8302)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(8299..8321)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 8310..8326
/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(8334..8364)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 8379..8400
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
misc_RNA 8443..8518
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
terminator 8540..8574
/label=lambda t0 terminator
/note="minimal transcription terminator from phage lambda
(Scholtissek and Grosse, 1987)"
rep_origin complement(8616..9204)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(8696..8715)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS 9408..10208
/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 cggacgctcg gactgggcgc gggattccag cagtaaccca ggtccgccgg ccacctcacg
61 gcaggcagac cctcggcttt cgcgccggga ccgccatgag accgccaccc ggatgtccgg
121 ggtggcggtc tcatggcggt ccctcagcgg ccctacgcga ccgctgtgtc gacgcggagg
181 cagtctccgg ggtgctggcc ccagggctgg agtaccgggg cgagcttgcc cttgcagcgg
241 cggcacacgg gcgacgcggc gccgggcggg ggagtcttga cgtcgacggt gaccggcttc
301 ggcttcaggc gcttccgaag gctgatcgtc gggcggatcg cctccttctt cggctgacgc
361 actcggtcga gcgaagcggc cagctcctcc gtgcgggcct cgttggcctt ccggcgcgcc
421 tcgcggaaag cagcttcctc ctcggacatg acctcgaacc tcatctggtc agcgtcaagg
481 tcgcccggcg cggccggcgc ttccggggcg ggcgggtcca ggacgtcctt gccccacacc
541 aggccccagg actcgacgag ccgccggacg cccggtaggc cgtacgtctc ggcgaccttg
601 atgaggtcga ggcgacgtcc ggcgacgcgg gcgatgtatc ggtaccagat gtaggccggg
661 atgaccgcga tggcgaccag gccctcggtg tcgtcggtga tctcctcctc ggtgcggacg
721 tcctgctgga tgccgagttc cttgatcagc cggttcaggt tctgcgaccg gtagtgcttg
781 cggacctgga acacgccgaa ctcgcgctcg cggtacttct cgacgaacgg gccgggccga
841 cgaagccgct gcagctcggc ggccgccgcg tcgcccaggt cgagcggtcc catgcggtcg
901 tcgccgcgac cggccttgaa gttctgtccg gccagctcca ggccgatctt ggcgacgccg
961 cccttggtct tgtcgccgtc cttgtagagg tagcgggcct gcttgcccgc atcgccgtca
1021 gcggcgtccg cgccgttgag tgggcgcacg tcggtgccgt ggcccttgcc ctcacaggag
1081 caaccgggcc ggtcgcacgt ctcgctgacg gtgtagccgc ccgcggattc gaccccggcg
1141 gcccaggctc cggcgagtgc gtcgcggaac gcggcctggg cgtccgggcc gagcacctcg
1201 cgggtgaccc agagcgtgtg ccagtgcagg tgccagccgg agccccagcc gaaggtgtcc
1261 tcgaaggccc gctcgtagcc gatgatcccg aagtcgtcgc gcatcgtgcg ccagcggcgg
1321 ccggacgagc cgtacgcgcc cttccagccg tcgtgcaaga ccgcgaccag gccgtgccgc
1381 attcccttgc ggacggtgcc gaacgccatg cgctcgaagt ggcgcaacgt gttcgtgcca
1441 aggtgcagcc cgtacccggc gtccgcgaga ccgtcggcgg cgagctgcac gttcgagccc
1501 cgtacggcca ggatgcggct catgcaccac gggcaggtgt ggacgttgtt gcagcggcac
1561 gtgttgcccc acgtcgcctc gcccggcttc cacatcagct cggccgtccc ggcagtgagc
1621 cgggtcccgc agcccttgaa cgcctcgttc agcgacaccg tctggtgccg gtcccgccgg
1681 gcgaaccgct cgtcgcgcgg gtcctcccgc ccggctgtcg cggcaccctc gtttggggta
1741 gaacccgttc cagttacagc gctctgacct gcagtggacg gagattttcc cttactacta
1801 aagcccgcgt ccggattacc cgctgtagtc gtgcttgcta cgctgcgtga ctggtccgca
1861 atgagacgct ttgcgcgctt tcggcaggcg tccgagcagt agattttggg gcgcttcccg
1921 gggatgtgga cgatcggggt gccgcagtgg cacttcggtc cggcggggcg cggtggtgtc
1981 gacgcgctgt tctctcgtac gctcgtcaca gagcaaacgt cctcactcgg catgctgcgc
2041 cggttcgggg gcggcgagcc cgggaggcca atcccgggct cgtgccattt ctgggtcctg
2101 ttgatcctgg cattggtgtg gccgttcatt gcccctgctc gctcctgacg cgccgataga
2161 cgtccgatac gcccggtgct ggtgggattt gataggtcgg aagaagcccc gcccgggcct
2221 gggcggggct tcctgtgcgt caggacctcc tcgtcgtgag cctcttcggc ctatggacgg
2281 agtgacctcg tgatccgtta cagccgcgcg cgctcgcgta gagcggtctc atcagttcca
2341 cgaacggtcc tcttcgcaga tcagggcgtt ggggcggagt ctcaccaagg actacgtctg
2401 ctggcgattt ccgttacacc ccgggcggtg gccggcgcac acgcgcgccc gcgttgggca
2461 gtgcagaaag tgcagaaacc taggcgctga tggtccaggt ccacggttcg tcgtcggcgg
2521 cggcgcgggc ggcggcgtcg gccagggcgc gggcgagacc ggctacggcg ggcttgatgc
2581 gccggttgcg ggcgaccttg agcagctagt atgcaggtcg acggatcttt tccgctgcat
2641 aaccctgctt cggggtcatt atagcgattt tttcggtata tccatccttt ttcgcacgat
2701 atacaggatt ttgccaaagg gttcgtgtag actttccttg gtgtatccaa cggcgtcagc
2761 cgggcaggat aggtgaagta ggcccacccg cgagcgggtg ttccttcttc actgtccctt
2821 attcgcacct ggcggtgctc aacgggaatc ctgctctgcg aggctggccg gctaccgccg
2881 gcgtaacaga tgagggcaag cggatggctg atgaaaccaa gccaaccagg aagggcagcc
2941 cacctatcaa ggtgtactgc cttccagacg aacgaagagc gattgaggaa aaggcggcgg
3001 cggccggcat gagcctgtcg gcctacctgc tggccgtcgg ccagggctac aaaatcacgg
3061 gcgtcgtgga ctatgagcac gtccgcgagc tggccctgca ggcggaagtc aggtagacac
3121 gacttccgct agtccttgca aggtctgctg acgtgaggcg gggcggtcgt ttttgaccgc
3181 cctgccttcg tcatgtaggc tcgctcgctg tgcctggcgt gtcatcagac gcccaggtcc
3241 cggtgccgtg aggcccgggc catcgagccg gtggtacgtg gctgcggtcc ccttgtgagg
3301 gctgcgcgcc gtgtgctgtc cggcgcgcac agccttgaat ccacccgcgg gggccggccg
3361 gtctccgtga gctcgagtag acgacggaga cgtaatggac aagaagtaca gcatcggcct
3421 ggacatcggc accaacagcg tgggctgggc ggtcatcacc gacgagtaca aggtcccctc
3481 caagaagttc aaggtcctgg gcaacaccga ccggcactcg atcaagaaga acctgatcgg
3541 cgccctgctc ttcgacagcg gcgaaaccgc cgaggcgacc cgcctgaagc ggaccgcccg
3601 tcgccgctac acccggcgca agaaccgcat ctgctacctg caggagatct tctccaacga
3661 gatggccaag gtcgacgact cgttcttcca ccggctcgag gagagcttcc tggtggagga
3721 ggacaagaag cacgagcgcc acccgatctt cggcaacatc gtcgacgagg tggcctacca
3781 cgagaagtac cccaccatct accacctccg caagaagctg gtcgactcga ccgacaaggc
3841 ggacctgcgg ctcatctacc tggccctcgc gcacatgatc aagttccgcg gccacttcct
3901 catcgagggc gacctgaacc cggacaactc cgacgtcgac aagctcttca tccagctggt
3961 gcagacctac aaccagctgt tcgaggagaa ccccatcaac gccagcggcg tcgacgccaa
4021 ggcgatcctc tccgcgcgcc tgagcaagtc ccggcgcctg gagaacctca tcgcccagct
4081 gccgggcgag aagaagaacg gcctcttcgg caacctgatc gcgctgtcgc tcggcctgac
4141 ccccaacttc aagagcaact tcgacctggc cgaggacgcg aagctccagc tgtccaagga
4201 cacctacgac gacgacctgg acaacctgct cgcccagatc ggcgaccagt acgcggacct
4261 cttcctggcc gcgaagaacc tctcggacgc catcctgctc agcgacatcc tgcgggtcaa
4321 caccgagatc accaaggccc cgctgtcggc gagcatgatc aagcggtacg acgagcacca
4381 ccaggacctg accctgctca aggccctcgt gcgccagcag ctgcccgaga agtacaagga
4441 gatcttcttc gaccagtcca agaacggcta cgccggctac atcgacggcg gcgcgtcgca
4501 ggaggagttc tacaagttca tcaagccgat cctggagaag atggacggca ccgaggagct
4561 gctcgtcaag ctgaaccgcg aggacctgct ccgcaagcag cggaccttcg acaacggctc
4621 catcccgcac cagatccacc tgggcgagct ccacgccatc ctccggcgcc aggaggactt
4681 ctaccccttc ctgaaggaca accgcgagaa gatcgagaag atcctgacct tccggatccc
4741 gtactacgtc ggccccctgg cccgcggcaa ctcccggttc gcgtggatga cccggaagtc
4801 ggaggaaacc atcaccccgt ggaacttcga ggaggtcgtg gacaagggcg cctccgcgca
4861 gtcgttcatc gagcgcatga ccaacttcga caagaacctc ccgaacgaga aggtcctgcc
4921 caagcacagc ctgctctacg agtacttcac cgtgtacaac gagctgacca aggtcaagta
4981 cgtgaccgag ggcatgcgga agccggcctt cctgtccggc gagcagaaga aggcgatcgt
5041 cgacctgctc ttcaagacca accgcaaggt caccgtgaag cagctgaagg aggactactt
5101 caagaagatc gagtgcttcg actccgtcga gatctcgggc gtggaggacc gcttcaacgc
5161 ctccctgggc acctaccacg acctgctcaa gatcatcaag gacaaggact tcctcgacaa
5221 cgaggagaac gaggacatcc tggaggacat cgtcctcacc ctgaccctct tcgaggaccg
5281 cgagatgatc gaggagcggc tcaagaccta cgcccacctg ttcgacgaca aggtgatgaa
5341 gcagctgaag cggcgccggt acaccggctg gggccgcctc tcccggaagc tgatcaacgg
5401 catccgggac aagcagagcg gcaagaccat cctggacttc ctcaagtccg acggcttcgc
5461 caaccgcaac ttcatgcagc tcatccacga cgactcgctg accttcaagg aggacatcca
5521 gaaggcccag gtgtccggcc agggcgacag cctccacgag cacatcgcca acctggcggg
5581 ctccccggcg atcaagaagg gcatcctcca gaccgtcaag gtcgtggacg agctggtcaa
5641 ggtgatgggc cgccacaagc ccgagaacat cgtgatcgag atggcccggg agaaccagac
5701 cacccagaag ggccagaaga actcccgcga gcggatgaag cgcatcgagg agggcatcaa
5761 ggagctcggc tcgcagatcc tgaaggagca cccggtcgag aacacccagc tccagaacga
5821 gaagctgtac ctctactacc tgcagaacgg ccgcgacatg tacgtggacc aggagctcga
5881 catcaaccgg ctgagcgact acgacgtcga ccacatcgtg ccgcagtcct tcctgaagga
5941 cgactcgatc gacaacaagg tcctgacccg ctccgacaag aaccggggca agtccgacaa
6001 cgtgccctcg gaggaggtcg tgaagaagat gaagaactac tggcgccagc tgctcaacgc
6061 caagctcatc acccagcgca agttcgacaa cctgaccaag gccgagcggg gcggcctgtc
6121 ggagctcgac aaggcgggct tcatcaagcg ccagctcgtc gaaacccggc agatcaccaa
6181 gcacgtggcc cagatcctgg acagccggat gaacaccaag tacgacgaga acgacaagct
6241 gatccgcgag gtcaaggtga tcaccctcaa gagcaagctg gtgtccgact tccgcaagga
6301 cttccagttc tacaaggtcc gggagatcaa caactaccac cacgcccacg acgcgtacct
6361 gaacgccgtc gtgggcaccg cgctgatcaa gaagtacccg aagctggagt ccgagttcgt
6421 ctacggcgac tacaaggtct acgacgtgcg caagatgatc gccaagtcgg agcaggagat
6481 cggcaaggcc accgcgaagt acttcttcta cagcaacatc atgaacttct tcaagaccga
6541 gatcaccctg gccaacggcg agatccgcaa gcggcccctg atcgaaacca acggcgaaac
6601 cggcgagatc gtctgggaca agggccgcga cttcgccacc gtccggaagg tgctgtccat
6661 gccgcaggtc aacatcgtca agaaaaccga ggtgcagacc ggcggcttca gcaaggagtc
6721 catcctcccc aagcgcaact cggacaagct gatcgcccgg aagaaggact gggacccgaa
6781 gaagtacggc ggcttcgaca gccccaccgt cgcctactcc gtgctggtcg tggcgaaggt
6841 cgagaagggc aagagcaaga agctgaagtc cgtgaaggag ctgctcggca tcaccatcat
6901 ggagcgctcc tcgttcgaga agaacccgat cgacttcctg gaggccaagg gctacaagga
6961 ggtcaagaag gacctcatca tcaagctgcc caagtactcg ctgttcgagc tcgagaacgg
7021 ccgcaagcgg atgctcgcca gcgcgggcga gctgcagaag ggcaacgagc tggccctccc
7081 gtccaagtac gtcaacttcc tgtacctcgc gtcccactac gagaagctga agggctcgcc
7141 cgaggacaac gagcagaagc agctcttcgt ggagcagcac aagcactacc tggacgagat
7201 catcgagcag atctcggagt tcagcaagcg ggtcatcctg gccgacgcga acctcgacaa
7261 ggtgctgtcc gcctacaaca agcaccgcga caagccgatc cgggagcagg cggagaacat
7321 catccacctg ttcaccctca ccaacctggg cgcccccgcc gcgttcaagt acttcgacac
7381 caccatcgac cgcaagcggt acaccagcac caaggaggtc ctcgacgcga ccctgatcca
7441 ccagtccatc accggcctgt acgaaacccg catcgacctc tcccagctcg gcggcgactg
7501 agaattcaga tctacgcgtt ccccgcaaaa gcggcctttg actccctgca agcctcagcg
7561 accgaatata tcggttatgc gtgggcgatg gttgttgtca ttgtcggcgc aactatcggt
7621 atcaagctgt ttaagaaatt cacctcgaaa gcaagctgat aaaccgatac aattaaaggc
7681 tccttttgga gccttttttt gctgctcctt cggtcggacg tgcgtctacg ggcaccttac
7741 cgcagccgtc ggctgtgcga cacggacgga tcgggcgaac tggccgatgc tgggagaagc
7801 gcgctgctgt acggcgcgca ccgggtgcgg agcccctcgg cgagcggtgt gaaacttctg
7861 tgaatggcct gttcggttgc tttttttata cggctgccag ataaggcttg cagcatctgg
7921 gcggctaccg ctatgatcgg ggcgttcctg caattcttag tgcgagtatc tgaaagggga
7981 tacgcaagtc ttctcagccg ctacagggcg cgtcccattc gccattcagg ctgcgcaact
8041 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
8101 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
8161 cgacggccag tgagcgcgcg taatacgact cactataggg cgaattgggt accgggcccc
8221 ccctcgaggt cctccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa
8281 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
8341 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
8401 attgcgttgc gctcactgcc cgctttccac cggtgaagac atgttttaga gctagaaata
8461 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
8521 tttactccat ctggatttgt tcagaacgct cggttgccgc cgggcgtttt ttatctagag
8581 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
8641 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
8701 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
8761 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
8821 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgacccc
8881 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
8941 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
9001 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
9061 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
9121 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
9181 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
9241 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
9301 acctagatcc ttttggttca tgtgcagctc catcagcaaa aggggatgat aagtttatca
9361 ccaccgacta tttgcaacag tgccgttgat cgtgctatga tcgactgatg tcatcagcgg
9421 tggagtgcaa tgtcgtgcaa tacgaatggc gaaaagccga gctcatcggt cagcttctca
9481 accttggggt tacccccggc ggtgtgctgc tggtccacag ctccttccgt agcgtccggc
9541 ccctcgaaga tgggccactt ggactgatcg aggccctgcg tgctgcgctg ggtccgggag
9601 ggacgctcgt catgccctcg tggtcaggtc tggacgacga gccgttcgat cctgccacgt
9661 cgcccgttac accggacctt ggagttgtct ctgacacatt ctggcgcctg ccaaatgtaa
9721 agcgcagcgc ccatccattt gcctttgcgg cagcggggcc acaggcagag cagatcatct
9781 ctgatccatt gcccctgcca cctcactcgc ctgcaagccc ggtcgcccgt gtccatgaac
9841 tcgatgggca ggtacttctc ctcggcgtgg gacacgatgc caacacgacg ctgcatcttg
9901 ccgagttgat ggcaaaggtt ccctatgggg tgccgagaca ctgcaccatt cttcaggatg
9961 gcaagttggt acgcgtcgat tatctcgaga atgaccactg ctgtgagcgc tttgccttgg
10021 cggacaggtg gctcaaggag aagagccttc agaaggaagg tccagtcggt catgcctttg
10081 ctcggttgat ccgctcccgc gacattgtgg cgacagccct gggtcaactg ggccgagatc
10141 cgttgatctt cctgcatccg ccagaggcgg gatgcgaaga atgcgatgcc gctcgccagt
10201 cgattggctg agctcatgag cggagaacga gatgacgttg gaggggcaag gtcgcgctga
10261 ttgctggggc aacacgtgga gcggatcggg gattgtcttt cttcagctcg ctgatgatat
10321 gctgacgctc aatgccgttt ggcctccgac taacgaaaat cccgcatttg gacggctgat
10381 ccgattggca cggcggacgg cgaatggcgg agcagacgct cgtccggggg caatgagata
10441 tgaaaaagcc tgaactcacc gcgacgtatc gatgtcgagg ttcctcaggg gagccacccc
10501 agagaagccc tcggagctga gcggagctat ttccaaagcc atgccagcta gagacagtgc
10561 acaccgccaa gcatctgcaa aaccctcgcc tggagggaaa gtgcaatgta cgtacgctgg
10621 tttccctcca gaaagggatt ctgcggcttc tcaactttgg aagaagaggc ggaacgaact
10681 gctgtttcag cctactcatg tgaggaggct ggtcctttac cctgatgccc gagggcatca
10741 gggtaaagga ccagcctcgc caactaggag cgctaccaag gcgaacacaa cccaggactt
10801 ggtagaaccc ttcgccggag aagttcagac tcatcggcat gagcacccct cgccgcgcac
10861 tcggcaccgg tccgtcgacc accaccagga cgtcgttgtc gacgtcggcc ccgcggctcc
10921 tgcccgccga acgcgtcgtc gtcgacggcc tggtgctcat cgacgagcac ccggagccag
10981 gtgaaaagcg ccgg
//
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