| 质粒名称: | p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A |
|---|---|
| 出品公司: | 爱迪基因 |
| 目录编号: | 91861 |
| 存储实验室: | Jennifer Doudna |
|
载体骨架基本信息 |
|
| 载体名称: | p2CT |
| 载体出品公司: | UC Berkeley Macrolab |
| 载体抗性: | 氨苄青霉素 |
| 空载体大小: | -- |
| 完整质粒大小: | -- |
| 载体修饰: | -- |
| 载体类型: | 细菌表达 |
| 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: | LbuCas13a (LbuC2c2) R0179A/K1080A |
| 物种来源: | -- |
| 插入大小: | -- |
| 基因突变: | K1080A |
| GenBank编号 | -- |
| 启动子 | -- |
| 标签/融合蛋白: | His6-MBP-TEV site（主干上的N终端） |
| 克隆方法: | Ligation Independent克隆 |
| 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: | p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A was a gift from Jennifer Doudna (Addgene plasmid # 91861 ; http://n2t.net/addgene:91861 ; RRID:Addgene_91861) |
| 参考文献部分: | RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas Enzymes. East-Seletsky A, O'Connell MR, Burstein D, Knott GJ, Doudna JA. Mol Cell. 2017 May 4;66(3):373-383.e3. doi: 10.1016/j.molcel.2017.04.008. 10.1016/j.molcel.2017.04.008 PubMed 28475872 |
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 p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A 9434 bp ds-DNA circular SYN 06-AUG-2018
DEFINITION Bacterial expression for Cas13a.
ACCESSION .
VERSION .
KEYWORDS p2CT-His-MBP-Lbu_C2c2_R1079A_K1080A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9434)
AUTHORS East-Seletsky A, O'Connell MR, Burstein D, Knott GJ, Doudna JA
TITLE RNA Targeting by Functionally Orthogonal Type VI-A CRISPR-Cas
Enzymes.
JOURNAL Mol Cell. 2017 May 4;66(3):373-383.e3. doi:
10.1016/j.molcel.2017.04.008.
PUBMED 28475872
REFERENCE 2 (bases 1 to 9434)
AUTHORS .
TITLE Direct Submission
JOURNAL Exported Aug 6, 2018 from SnapGene Server 1.1.58
http://www.snapgene.com
FEATURES Location/Qualifiers
source 1..9434
/organism="synthetic DNA construct"
/mol_type="other DNA"
primer_bind 118..137
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 371..388
/label=L4440
/note="L4440 vector, forward primer"
misc_feature 403..542
/label=bom
/note="basis of mobility region from pBR322"
primer_bind complement(468..487)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 628..650
/label=pGEX 3'
/note="pGEX vectors, reverse primer"
CDS complement(644..835)
/codon_start=1
/gene="rop"
/product="Rop protein, which maintains plasmids at low copy
number"
/label=rop
/translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
DELYRSCLARFGDDGENL"
primer_bind 2293..2312
/label=pBRrevBam
/note="pBR322 vectors, tet region, downstream of BamHI,
reverse primer"
primer_bind 2396..2415
/label=T7
/note="T7 promoter, forward primer"
promoter 2396..2414
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
RBS 2503..2525
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 2545..2562
/codon_start=1
/product="6xHis affinity tag"
/label=6xHis
/translation="HHHHHH"
CDS 2572..3672
/codon_start=1
/gene="malE (mutated)"
/product="maltose binding protein from E. coli"
/label=MBP
/note="This version of the gene does not encode a signal
sequence, so MBP will remain in the cytosol."
/translation="MKIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLE
EKFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKL
IAYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIA
ADGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGE
TAMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFL
ENYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAF
WYAVRTAVINAASGRQTVDEALKDAQT"
primer_bind 3646..3669
/label=MBP-F
/note="Maltose binding protein, forward primer"
CDS 3727..3747
/codon_start=1
/product="tobacco etch virus (TEV) protease recognition and
cleavage site"
/label=TEV site
/translation="ENLYFQS"
primer_bind complement(7359..7377)
/label=T7 Term
/note="T7 terminator, reverse primer"
terminator 7373..7420
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
promoter complement(7786..7814)
/label=tet promoter
/note="E. coli promoter for tetracycline efflux protein
gene"
primer_bind complement(7841..7859)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 7927..8031
/gene="bla"
/label=AmpR promoter
CDS 8032..8892
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind complement(8250..8269)
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
rep_origin 9063..217
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
61 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
121 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
181 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
241 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
301 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
361 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct
421 ccttacgcat ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct
481 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
541 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
601 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
661 catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt
721 cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg
781 tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg
841 cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga
901 tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta
961 aacaactggc ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc
1021 gcttcgttaa tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga
1081 tccggaacat aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga
1141 aaccgaagac cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc
1201 acgttcgctc gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta
1261 gccgggtcct caacgacagg agcacgatca tgcgcacccg tggccaggac ccaacgctgc
1321 ccgagatgcg ccgcgtgcgg ctgctggaga tggcggacgc gatggatatg ttctgccaag
1381 ggttggtttg cgcattcaca gttctccgca agaattgatt ggctccaatt cttggagtgg
1441 tgaatccgtt agcgaggtgc cgccggcttc cattcaggtc gaggtggccc ggctccatgc
1501 accgcgacgc aacgcgggga ggcagacaag gtatagggcg gcgcctacaa tccatgccaa
1561 cccgttccat gtgctcgccg aggcggcata aatcgccgtg acgatcagcg gtccaatgat
1621 cgaagttagg ctggtaagag ccgcgagcga tccttgaagc tgtccctgat ggtcgtcatc
1681 tacctgcctg gacagcatgg cctgcaacgc gggcatcccg atgccgccgg aagcgagaag
1741 aatcataatg gggaaggcca tccagcctcg cgtcgcgaac gccagcaaga cgtagcccag
1801 cgcgtcggcc gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg
1861 accagtgacg aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc
1921 gatcatcgtc gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg
1981 cacctgtcct acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat
2041 gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgacg
2101 ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga ggccgttgag
2161 caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc ccccggccac
2221 ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg
2281 atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt
2341 gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc gaaattaata
2401 cgactcacta tagggagacc acaacggttt ccctctagtg ccggctccgg agagctcttt
2461 aattaagcgg ccgccctgca ggactcgagt tctagaaata attttgttta actttaagaa
2521 ggagatatac atatgaaatc ttctcaccat caccatcacc atggttcttc tatgaaaatc
2581 gaagaaggta aactggtaat ctggattaac ggcgataaag gctataacgg tctcgctgaa
2641 gtcggtaaga aattcgagaa agataccgga attaaagtca ccgttgagca tccggataaa
2701 ctggaagaga aattcccaca ggttgcggca actggcgatg gccctgacat tatcttctgg
2761 gcacacgacc gctttggtgg ctacgctcaa tctggcctgt tggctgaaat caccccggac
2821 aaagcgttcc aggacaagct gtatccgttt acctgggatg ccgtacgtta caacggcaag
2881 ctgattgctt acccgatcgc tgttgaagcg ttatcgctga tttataacaa agatctgctg
2941 ccgaacccgc caaaaacctg ggaagagatc ccggcgctgg ataaagaact gaaagcgaaa
3001 ggtaagagcg cgctgatgtt caacctgcaa gaaccgtact tcacctggcc gctgattgct
3061 gctgacgggg gttatgcgtt caagtatgaa aacggcaagt acgacattaa agacgtgggc
3121 gtggataacg ctggcgcgaa agcgggtctg accttcctgg ttgacctgat taaaaacaaa
3181 cacatgaatg cagacaccga ttactccatc gcagaagctg cctttaataa aggcgaaaca
3241 gcgatgacca tcaacggccc gtgggcatgg tccaacatcg acaccagcaa agtgaattat
3301 ggtgtaacgg tactgccgac cttcaagggt caaccatcca aaccgttcgt tggcgtgctg
3361 agcgcaggta ttaacgccgc cagtccgaac aaagagctgg caaaagagtt cctcgaaaac
3421 tatctgctga ctgatgaagg tctggaagcg gttaataaag acaaaccgct gggtgccgta
3481 gcgctgaagt cttacgagga agagttggcg aaagatccac gtattgccgc cactatggaa
3541 aacgcccaga aaggtgaaat catgccgaac atcccgcaga tgtccgcttt ctggtatgcc
3601 gtgcgtactg cggtgatcaa cgccgccagc ggtcgtcaga ctgtcgatga agccctgaaa
3661 gacgcgcaga ctaattcgag ctcgaacaac aacaacaata acaataacaa caacctcggg
3721 atcgaggaaa acctgtactt ccaatccaat gcaatgaaag tgacgaaggt aggaggcatt
3781 tcgcataaga agtacacgtc cgaaggccgc ttagtgaagt cagaatcgga agaaaatcgc
3841 acagacgaac gtctgtcggc gttgcttaat atgcgccttg acatgtatat caagaatccc
3901 agcagcacgg aaaccaagga aaatcaaaaa cgcattggga aattaaagaa attcttctca
3961 aacaaaatgg tctatcttaa agacaatacc ttgagtttga agaatgggaa aaaggagaac
4021 attgatcgtg agtattctga gactgacatc cttgagagcg atgtccgtga caagaaaaac
4081 ttcgccgtgt tgaaaaagat ctatctgaat gaaaacgtga actcggagga attggaagtt
4141 tttcgtaacg acattaagaa gaaactgaac aaaatcaaca gcctgaagta ctcatttgaa
4201 aagaataagg cgaattatca aaagattaat gagaataaca tcgagaaggt tgaaggtaag
4261 tcaaagcgta acattattta cgattattat cgtgagtcag cgaaacgtga cgcttatgta
4321 agcaatgtga aagaagcctt tgataagctt tacaaggaag aggacattgc aaaacttgtt
4381 cttgaaattg agaaccttac gaagttagag aaatacaaga ttcgcgagtt ctaccacgaa
4441 attattggac gtaagaatga caaggaaaac tttgcaaaaa tcatctacga agaaatccag
4501 aatgttaata acatgaaaga gttgatcgag aaggtaccgg acatgagtga gttgaaaaag
4561 agccaagtat tttacaagta ttacttagac aaagaagagt tgaacgacaa gaacatcaaa
4621 tacgcgtttt gtcatttcgt ggaaatcgaa atgagtcagt tgctgaagaa ctacgtatat
4681 aagcgcttaa gtaatatctc gaatgacaaa attaagcgta tctttgaata ccagaacttg
4741 aaaaaattga tcgaaaataa gctgttaaac aaacttgaca cgtacgtccg taattgtgga
4801 aagtataatt attatttgca agacggcgaa attgccactt cagatttcat cgcccgcaac
4861 cgtcagaatg aagcgtttct tcgcaacatc attggggtgt catctgtggc ctacttttct
4921 cttcgcaaca ttcttgaaac ggagaacgag aatgatatta ctgggcgtat gcgcggcaaa
4981 acagttaaga acaataaagg tgaagagaag tacgtgtccg gagaagttga taagatctat
5041 aatgaaaata agaagaacga ggttaaggag aacttaaaaa tgttctattc gtacgatttc
5101 aatatggaca acaagaatga aatcgaagat ttcttcgcca acatcgacga ggcgatttct
5161 tccatccgtc acggtattgt ccacttcaac ttggaattag aaggtaagga tatctttgcg
5221 ttcaagaaca ttgcgccatc cgaaatctca aagaagatgt ttcagaatga gattaacgag
5281 aaaaaactga aattgaagat ctttcgtcaa ctgaactctg ccaacgtgtt ccgctatctc
5341 gaaaagtata aaattctgaa ttaccttaaa cgtacacgct tcgagtttgt caataaaaat
5401 atcccattcg tcccgtcttt caccaaatta tattcgcgca ttgatgacct gaagaatagt
5461 cttgggattt actggaaaac tccgaaaaca aacgacgaca ataagactaa ggagattatt
5521 gatgcccaaa tctatttgct taaaaacatc tattacgggg agttcctgaa ttatttcatg
5581 tcgaacaatg gtaatttctt tgagatttct aaagaaatca tcgaattgaa caagaacgat
5641 aaacgcaact taaagactgg gttttacaag ctgcaaaagt ttgaagacat ccaggagaag
5701 attccaaagg aatacttggc gaatatccag tccctgtaca tgattaatgc cggtaatcag
5761 gacgaagaag aaaaggacac ttatattgat ttcattcaaa agatcttctt aaagggattt
5821 atgacgtatc ttgctaataa cggtcgttta agtctgattt acatcggctc ggatgaagaa
5881 acaaatacgt cattagcaga aaagaagcaa gagtttgaca agttcttgaa gaagtacgag
5941 cagaacaata atatcaagat cccctatgag atcaatgaat tcctgcgtga gatcaaactg
6001 ggaaacatcc tgaagtatac tgagcgttta aacatgttct accttatctt aaagcttttg
6061 aatcacaagg agctgacaaa tctgaagggt agtcttgaaa aatatcagtc tgccaataag
6121 gaagaagcgt tctctgacca attggagtta attaacctgc ttaaccttga caacaaccgc
6181 gtgacggaag acttcgaatt agaggccgac gagattggaa aatttcttga tttcaatggc
6241 aacaaagtta aggataacaa ggaactgaaa aagttcgata caaacaagat ctactttgac
6301 ggcgagaaca ttatcaaaca ccgtgccttc tacaatatta agaaatatgg catgttaaac
6361 ttactggaga aaattgccga caaggctgga tacaagatct cgatcgaaga gctgaagaaa
6421 tactccaata aaaagaatga gatcgagaag aaccataaga tgcaggaaaa tctgcaccgc
6481 aaatacgctc gtccccgtaa agacgagaag tttacagatg aggactatga aagttacaag
6541 caagctattg agaatattga ggagtacacc caccttaaga acaaggtaga attcaatgag
6601 ctgaatttac tgcagggcct gttgctgcgc attttacatc gtttagtcgg atatacctca
6661 atttgggaac gcgatctgcg cttccgcctt aaaggtgagt tcccagaaaa ccaatacatc
6721 gaagagatct tcaactttga aaataagaag aacgtgaagt acaaaggggg tcagattgta
6781 gagaaataca ttaaattcta caaggaatta catcaaaatg atgaagttaa gatcaacaag
6841 tacagttccg cgaatatcaa ggtgttgaag caagaaaaga aggaccttta tattcgcaat
6901 tacatcgccc acttcaatta tattcctcac gccgagatct cactgctgga agtccttgaa
6961 aatttgcgta aattgctgtc ctacgatgcc gcactgaaaa atgccgtaat gaaatcagta
7021 gttgatatcc ttaaggagta tggttttgta gccacattca aaatcggggc ggacaagaag
7081 atcggtattc agacactgga gagcgaaaaa atcgtgcatc ttaagaatct taagaagaag
7141 aagttaatga ctgaccgcaa ttccgaggaa ctttgcaaat tggtgaagat tatgtttgaa
7201 tacaaaatgg aagagaaaaa gtctgaaaac taataacatt ggaagtggat aacggatccg
7261 cgatcgcggc gcgccacctg gtggccggcc ggtaccacgc gtgcgcgctg atccggctgc
7321 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
7381 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
7441 cggatatcca caggacgggt gtggtcgcca tgatcgcgta gtcgatagtg gctccaagta
7501 gcgaagcgag caggactggg cggcggccaa agcggtcgga cagtgctccg agaacgggtg
7561 cgcatagaaa ttgcatcaac gcatatagcg ctagcagcac gccatagtga ctggcgatgc
7621 tgtcggaatg gacgatatcc cgcaagaggc ccggcagtac cggcataacc aagcctatgc
7681 ctacagcatc cagggtgacg gtgccgagga tgacgatgag cgcattgtta gatttcatac
7741 acggtgcctg actgcgttag caatttaact gtgataaact accgcattaa agcttatcga
7801 tgataagctg tcaaacatga gaattcttga agacgaaagg gcctcgtgat acgcctattt
7861 ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga
7921 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
7981 atgagacaat aaccctgata aatgcttcaa taacattgaa aaaggaagag tatgagtatt
8041 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
8101 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
8161 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
8221 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
8281 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
8341 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
8401 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
8461 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
8521 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgcagca
8581 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
8641 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
8701 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
8761 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
8821 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
8881 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
8941 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
9001 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
9061 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
9121 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
9181 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
9241 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
9301 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
9361 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
9421 acctacaccg aact
//
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