| 质粒名称: | pCAS |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 60847 |
| 存储实验室: | Jamie Cate |
|
载体骨架基本信息 |
|
| 载体名称: | 2-micron/pUC |
| 载体出品公司: | -- |
| 载体抗性: | 卡那霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | 8761 bp |
| 载体修饰: | -- |
| 载体类型: | 细菌表达，酵母表达，CRISPR，合成生物学 |
| 筛选标记: | G418 |
| 高拷贝/低拷贝: | 高拷贝 |
| 生长条件: | 37 ℃ |
|
插入基因信息 |
|
| 插入基因名称: | S. pyogenese Cas9 |
| 物种来源: | 化脓性链球菌 |
| 插入大小: | 4104 bp |
| 基因突变: | -- |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | NLS/His8（插入件上的C端子） |
| 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: | pCAS was a gift from Jamie Cate (Addgene plasmid # 60847 ; http://n2t.net/addgene:60847 ; RRID:Addgene_60847) |
| 参考文献部分: | Selection of chromosomal DNA libraries using a multiplex CRISPR system. Ryan OW, Skerker JM, Maurer MJ, Li X, Tsai JC, Poddar S, Lee ME, DeLoache W, Dueber JE, Arkin AP, Cate JH. Elife. 2014 Aug 19;3. doi: 10.7554/eLife.03703. 10.7554/eLife.03703 PubMed 25139909 |
买家导航
pCAS载体质粒基本信息
pCAS质粒图谱载体图谱和pCAS载体序列质粒序列多克隆位点信息
pCAS质粒载体简介
pCAS质粒序列载体序列
LOCUS pCAS 8743 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Expresses S. pyogenes Cas9 plus an HDV ribozyme-sgRNA for genome
editing in yeast.
ACCESSION .
VERSION .
KEYWORDS pCAS
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8743)
AUTHORS Ryan OW, Skerker JM, Maurer MJ, Li X, Tsai JC, Poddar S, Lee ME,
DeLoache W, Dueber JE, Arkin AP, Cate JH
TITLE Selection of chromosomal DNA libraries using a multiplex CRISPR
system.
JOURNAL Elife. 2014 Aug 19;3. doi: 10.7554/eLife.03703.
PUBMED 25139909
REFERENCE 2 (bases 1 to 8743)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..8743
/organism="synthetic DNA construct"
/mol_type="other DNA"
protein_bind complement(749..796)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_RNA 1169..1244
/label=gRNA scaffold
/note="guide RNA scaffold for the Streptococcus pyogenes
CRISPR/Cas9 system"
gene 1378..2734
/label=kanMX
/note="yeast selectable marker conferring kanamycin
resistance (Wach et al., 1994)"
promoter 1378..1721
/label=TEF promoter
/note="Ashbya gossypii TEF promoter"
CDS 1722..2531
/codon_start=1
/gene="aph(3')-Ia"
/product="aminoglycoside phosphotransferase"
/label=KanR
/note="confers resistance to kanamycin"
/translation="MGKEKTHVSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
primer_bind complement(1789..1808)
/label=Kan-R
/note="Kanamycin resistance gene, reverse primer"
terminator 2537..2734
/label=TEF terminator
/note="Ashbya gossypii TEF terminator"
CDS 3497..7600
/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 7610..7630
/codon_start=1
/product="nuclear localization signal of SV40 (simian virus
40) large T antigen"
/label=SV40 NLS
/translation="PKKKRKV"
CDS 7646..7669
/codon_start=1
/product="8xHis affinity tag"
/label=8xHis
/translation="HHHHHHHH"
terminator 7673..7862
/gene="S. cerevisiae CYC1"
/label=CYC1 terminator
/note="transcription terminator for CYC1"
rep_origin complement(7919..8507)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(7999..8018)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
rep_origin 8529..768
/label=2u ori
/note="yeast 2u plasmid origin of replication"
ORIGIN
1 gaacagaaat gcaacgcgag agcgctattt taccaacaaa gaatctatac ttcttttttg
61 ttctacaaaa atgcatcccg agagcgctat ttttctaaca aagcatctta gattactttt
121 tttctccttt gtgcgctcta taatgcagtc tcttgataac tttttgcact gtaggtccgt
181 taaggttaga agaaggctac tttggtgtct attttctctt ccataaaaaa agcctgactc
241 cacttcccgc gtttactgat tactagcgaa gctgcgggtg cattttttca agataaaggc
301 atccccgatt atattctata ccgatgtgga ttgcgcatac tttgtgaaca gaaagtgata
361 gcgttgatga ttcttcattg gtcagaaaat tatgaacggt ttcttctatt ttgtctctat
421 atactacgta taggaaatgt ttacattttc gtattgtttt cgattcactc tatgaatagt
481 tcttactaca atttttttgt ctaaagagta atactagaga taaacataaa aaatgtagag
541 gtcgagttta gatgcaagtt caaggagcga aaggtggatg ggtaggttat atagggatat
601 agcacagaga tatatagcaa agagatactt ttgagcaatg tttgtggaag cggtattcgc
661 aatattttag tagcccgtta cagtccggtg cgtttttggt tttttgaaag tgcgtcttca
721 gagcgctttt ggttttcaaa agcgctctga agttcctata ctttctagag aataggaact
781 tcggaatagg aacttcaaag cgtttccgaa aacgagcgct tccgaaaatg caacgcgagc
841 tgcgcacata cagctcactg ttcacgtcgc acctatatct gcgtgttgcc tgtatatata
901 tatacatgag aagaacggca tagtgcgtgt ttatgcttaa atgcgtatat gtgttatgta
961 gtatactctt tcttcaacaa ttaaatactc tcggtagcca agttggttta aggcgcaaga
1021 ctgtaattta tcactacgaa atcttgagat cgggcgttcg actcgccccc gggagagatg
1081 gccggcatgg tcccagcctc ctcgctggcg ccggctgggc aacaccttcg ggtggcgaat
1141 gggactttca taataacgtc caataaatgt tttagagcta gaaatagcaa gttaaaataa
1201 ggctagtccg ttatcaactt gaaaaagtgg caccgagtcg gtgctttttt tattttttgt
1261 cactattgtt atgtaaaatg ccacctctga cagtatggaa cgcaaacttc tgtctagtgg
1321 atataggtct agagatctgt ttagcttgcc tcgtccccgc cgggtcaccc ggccagcgac
1381 atggaggccc agaataccct ccttgacagt cttgacgtgc gcagctcagg ggcatgatgt
1441 gactgtcgcc cgtacattta gcccatacat ccccatgtat aatcatttgc atccatacat
1501 tttgatggcc gcacggcgcg aagcaaaaat tacggctcct cgctgcagac ctgcgagcag
1561 ggaaacgctc ccctcacaga cgcgttgaat tgtccccacg ccgcgcccct gtagagaaat
1621 ataaaaggtt aggatttgcc actgaggttc ttctttcata tacttccttt taaaatcttg
1681 ctaggataca gttctcacat cacatccgaa cataaacaac catgggtaag gaaaagactc
1741 acgtttcgag gccgcgatta aattccaaca tggatgctga tttatatggg tataaatggg
1801 ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg attgtatggg aagcccgatg
1861 cgccagagtt gtttctgaaa catggcaaag gtagcgttgc caatgatgtt acagatgaga
1921 tggtcagact aaactggctg acggaattta tgcctcttcc gaccatcaag cattttatcc
1981 gtactcctga tgatgcatgg ttactcacca ctgcgatccc cggcaaaaca gcattccagg
2041 tattagaaga atatcctgat tcaggtgaaa atattgttga tgcgctggca gtgttcctgc
2101 gccggttgca ttcgattcct gtttgtaatt gtccttttaa cagcgatcgc gtatttcgtc
2161 tcgctcaggc gcaatcacga atgaataacg gtttggttga tgcgagtgat tttgatgacg
2221 agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat gcataagctt ttgccattct
2281 caccggattc agtcgtcact catggtgatt tctcacttga taaccttatt tttgacgagg
2341 ggaaattaat aggttgtatt gatgttggac gagtcggaat cgcagaccga taccaggatc
2401 ttgccatcct atggaactgc ctcggtgagt tttctccttc attacagaaa cggctttttc
2461 aaaaatatgg tattgataat cctgatatga ataaattgca gtttcatttg atgctcgatg
2521 agtttttcta atcagtactg acaataaaaa gattcttgtt ttcaagaact tgtcatttgt
2581 atagtttttt tatattgtag ttgttctatt ttaatcaaat gttagcgtga tttatatttt
2641 ttttcgcctc gacatcatct gcccagatgc gaagttaagt gcgcagaaag taatatcatg
2701 cgtcaatcgt atgtgaatgc tggtcgctat actgctgtcg attcgatact aacgccgcca
2761 tccagtgtcg aaaacgagct ctcgagaacc cttaatagtc gaacaagaag caggcaaagt
2821 ttagagcact gcccctccgc actcaaaaaa gaaaaaacta ggaggaaaat aaaattctca
2881 accacacaaa cacataaaca catacaaata caaatacaag cttatttact tgacatcgcg
2941 cgatcttcca ctattcagcg ccgtccgccc tctctcgtgt tttttgttta cgcgacaact
3001 atgcgaaatc cggagcaacg ggcaaccgtt tggggaaaga ccacacccac gcgcgatcgc
3061 catggcaacg aggtcgcaca cgccccacac ccagacctcc ctgcgagcgg gcatgggtac
3121 aatgtccccg ttgccacaga caccacttcg tagcacagcg cagagcgtag cgtgttgttg
3181 ctgctgacaa aagaaaattt ttcttagcaa agcaaaggag gggaagcacg ggcagatagc
3241 accgtaccat acccttggaa actcgaaatg aacgaagcag gaaatgagag aatgagagtt
3301 ttgtaggtat atatagcggt agtgtttgcg cgttaccatc atcttctgga tctatctatt
3361 gttcttttcc tcatcacttt cccctttttc gctcttcttc ttgtctttta tttctttctt
3421 ttttttaatt gttccctcga ttggctatct accaaagaat ccaaacttaa tacacgtatt
3481 tatttgtcca attaccatgg ataagaaata ctcaataggc ttagatatcg gcacaaatag
3541 cgtcggatgg gcggtgatca ctgatgaata taaggttccg tctaaaaagt tcaaggttct
3601 gggaaataca gaccgccaca gtatcaaaaa aaatcttata ggggctcttt tatttgacag
3661 tggagagaca gcggaagcga ctcgtctcaa acggacagct cgtagaaggt atacacgtcg
3721 gaagaatcgt atttgttatc tacaggagat tttttcaaat gagatggcga aagtagatga
3781 tagtttcttt catcgacttg aagagtcttt tttggtggaa gaagacaaga agcatgaacg
3841 tcatcctatt tttggaaata tagtagatga agttgcttat catgagaaat atccaactat
3901 ctatcatctg cgaaaaaaat tggtagattc tactgataaa gcggatttgc gcttaatcta
3961 tttggcctta gcgcatatga ttaagtttcg tggtcatttt ttgattgagg gagatttaaa
4021 tcctgataat agtgatgtgg acaaactatt tatccagttg gtacaaacct acaatcaatt
4081 atttgaagaa aaccctatta acgcaagtgg agtagatgct aaagcgattc tttctgcacg
4141 attgagtaaa tcaagacgat tagaaaatct cattgctcag ctccccggtg agaagaaaaa
4201 tggcttattt gggaatctca ttgctttgtc attgggtttg acccctaatt ttaaatcaaa
4261 ttttgatttg gcagaagatg ctaaattaca gctttcaaaa gatacttacg atgatgattt
4321 agataattta ttggcgcaaa ttggagatca atatgctgat ttgtttttgg cagctaagaa
4381 tttatcagat gctattttac tttcagatat cctaagagta aatactgaaa taactaaggc
4441 tcccctatca gcttcaatga ttaaacgcta cgatgaacat catcaagact tgactctttt
4501 aaaagcttta gttcgacaac aacttccaga aaagtataaa gaaatctttt ttgatcaatc
4561 aaaaaacgga tatgcaggtt atattgatgg gggagctagc caagaagaat tttataaatt
4621 tatcaaacca attttagaaa aaatggatgg tactgaggaa ttattggtga aactaaatcg
4681 tgaagatttg ctgcgcaagc aacggacctt tgacaacggc tctattcccc atcaaattca
4741 cttgggtgag ctgcatgcta ttttgagaag acaagaagac ttttatccat ttttaaaaga
4801 caatcgtgag aagattgaaa aaatcttgac ttttcgaatt ccttattatg ttggtccatt
4861 ggcgcgtggc aatagtcgtt ttgcatggat gactcggaag tctgaagaaa caattacccc
4921 atggaatttt gaagaagttg tcgataaagg tgcttcagct caatcattta ttgaacgcat
4981 gacaaacttt gataaaaatc ttccaaatga aaaagtacta ccaaaacata gtttgcttta
5041 tgagtatttt acggtttata acgaattgac aaaggtcaaa tatgttactg aaggaatgcg
5101 aaaaccagca tttctttcag gtgaacagaa gaaagccatt gttgatttac tcttcaaaac
5161 aaatcgaaaa gtaaccgtta agcaattaaa agaagattat ttcaaaaaaa tagaatgttt
5221 tgatagtgtt gaaatttcag gagttgaaga tagatttaat gcttcattag gtacctacca
5281 tgatttgcta aaaattatta aagataaaga ttttttggat aatgaagaaa atgaagatat
5341 cttagaggat attgttttaa cattgacctt atttgaagat agggagatga ttgaggaaag
5401 acttaaaaca tatgctcacc tctttgatga taaggtgatg aaacagctta aacgtcgccg
5461 ttatactggt tggggacgtt tgtctcgaaa attgattaat ggtattaggg ataagcaatc
5521 tggcaaaaca atattagatt ttttgaaatc agatggtttt gccaatcgca attttatgca
5581 gctgatccat gatgatagtt tgacatttaa agaagacatt caaaaagcac aagtgtctgg
5641 acaaggcgat agtttacatg aacatattgc aaatttagct ggtagccctg ctattaaaaa
5701 aggtatttta cagactgtaa aagttgttga tgaattggtc aaagtaatgg ggcggcataa
5761 gccagaaaat atcgttattg aaatggcacg tgaaaatcag acaactcaaa agggccagaa
5821 aaattcgcga gagcgtatga aacgaatcga agaaggtatc aaagaattag gaagtcagat
5881 tcttaaagag catcctgttg aaaatactca attgcaaaat gaaaagctct atctctatta
5941 tctccaaaat ggaagagaca tgtatgtgga ccaagaatta gatattaatc gtttaagtga
6001 ttatgatgtc gatcacattg ttccacaaag tttccttaaa gacgattcaa tagacaataa
6061 ggtcttaacg cgttctgata aaaatcgtgg taaatcggat aacgttccaa gtgaagaagt
6121 agtcaaaaag atgaaaaact attggagaca acttctaaac gccaagttaa tcactcaacg
6181 taagtttgat aatttaacga aagctgaacg tggaggtttg agtgaacttg ataaagctgg
6241 ttttatcaaa cgccaattgg ttgaaactcg ccaaatcact aagcatgtgg cacaaatttt
6301 ggatagtcgc atgaatacta aatacgatga aaatgataaa cttattcgag aggttaaagt
6361 gattacctta aaatctaaat tagtttctga cttccgaaaa gatttccaat tctataaagt
6421 acgtgagatt aacaattacc atcatgccca tgatgcgtat ctaaatgccg tcgttggaac
6481 tgctttgatt aagaaatatc caaaacttga atcggagttt gtctatggtg attataaagt
6541 ttatgatgtt cgtaaaatga ttgctaagtc tgagcaagaa ataggcaaag caaccgcaaa
6601 atatttcttt tactctaata tcatgaactt cttcaaaaca gaaattacac ttgcaaatgg
6661 agagattcgc aaacgccctc taatcgaaac taatggggaa actggagaaa ttgtctggga
6721 taaagggcga gattttgcca cagtgcgcaa agtattgtcc atgccccaag tcaatattgt
6781 caagaaaaca gaagtacaga caggcggatt ctccaaggag tcaattttac caaaaagaaa
6841 ttcggacaag cttattgctc gtaaaaaaga ctgggatcca aaaaaatatg gtggttttga
6901 tagtccaacg gtagcttatt cagtcctagt ggttgctaag gtggaaaaag ggaaatcgaa
6961 gaagttaaaa tccgttaaag agttactagg gatcacaatt atggaaagaa gttcctttga
7021 aaaaaatccg attgactttt tagaagctaa aggatataag gaagttaaaa aagacttaat
7081 cattaaacta cctaaatata gtctttttga gttagaaaac ggtcgtaaac ggatgctggc
7141 tagtgccgga gaattacaaa aaggaaatga gctggctctg ccaagcaaat atgtgaattt
7201 tttatattta gctagtcatt atgaaaagtt gaagggtagt ccagaagata acgaacaaaa
7261 acaattgttt gtggagcagc ataagcatta tttagatgag attattgagc aaatcagtga
7321 attttctaag cgtgttattt tagcagatgc caatttagat aaagttctta gtgcatataa
7381 caaacataga gacaaaccaa tacgtgaaca agcagaaaat attattcatt tatttacgtt
7441 gacgaatctt ggagctcccg ctgcttttaa atattttgat acaacaattg atcgtaaacg
7501 atatacgtct acaaaagaag ttttagatgc cactcttatc catcaatcca tcactggtct
7561 ttatgaaaca cgcattgatt tgagtcagct aggaggtgac ggtggagggc caaaaaagaa
7621 aagaaaagtt gaagatgctt ctgctcatca tcaccatcac catcaccatt aaatccgctc
7681 taaccgaaaa ggaaggagtt agacaacctg aagtctaggt ccctatttat ttttttatag
7741 ttatgttagt attaagaacg ttatttatat ttcaaatttt tctttttttt ctgtacagac
7801 gcgtgtacgc atgtaacatt atactgaaaa ccttgcttga gaaggttttg ggacgctcga
7861 agtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
7921 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
7981 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
8041 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
8101 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
8161 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
8221 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
8281 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
8341 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
8401 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
8461 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
8521 tcttttctac cgaagcatct gtgcttcatt ttgtagaaca aaaatgcaac gcgagagcgc
8581 taatttttca aacaaagaat ctgagctgca tttttacaga acagaaatgc aacgcgaaag
8641 cgctatttta ccaacgaaga atctgtgctt catttttgta aaacaaaaat gcaacgcgag
8701 agcgctaatt tttcaaacaa agaatctgag ctgcattttt aca
//
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