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note
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2, 0.5 mM 2-ME, 6 μg BSA, 2.5 μg circular plasmid, 180 U FEN1 or 50 ng XPG, 25 μM test compound, and 5 pmol BVT substrate) contained in black 96-well plates. Reactions were incubated at rt for 90 min, stopped through the addition of 40 μL stop buffer (0.025% SDS, 12.5 mM EDTA) and fluorescence was measured using a Fluoroscan plate-reading fluorometer fitted with 485 nm excitation/538 nm emission filters
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17
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0029990093
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J.E. Tomassini, M.E. Davies, J.C. Hastings, R. Lingham, M. Mojena, S.L. Raghoobar, S.B. Singh, J.S. Tkacz, and M.A. Goetz Antimicrob. Agents Chemother. 40 1996 1189 1193
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Tomassini, J.E.1
Davies, M.E.2
Hastings, J.C.3
Lingham, R.4
Mojena, M.5
Raghoobar, S.L.6
Singh, S.B.7
Tkacz, J.S.8
Goetz, M.A.9
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22
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10644223896
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note
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3 followed by an alkyl halide. After heating to 80°C for 2-12 h, the reaction mixture was cooled and partitioned between water and EtOAc. The organic layer was evaporated to give the crude product, which was generally used without purification. The O-protecting group was removed by standard procedures
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25
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10644245439
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note
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General protocol for the synthesis of the furan amino esters: The substituted acetonitrile was added to a solution of ethyl formate (1.1 equiv) and sodium methoxide (1.1 equiv) in MeOH (0.1 M). After refluxing for 2 h, the resulting precipitate was filtered and washed with ether to give the crude hydroxyacrylonitrile. Diethylchloromalonate (1.0 equiv) was added to a stirred solution of the crude hydroxyacrylonitrile in DMF (0.1 M). After stirring 5 h at rt, the solvent was removed and the resulting oil was dissolved in ethanol. DBN (1.0 equiv) was added and the reaction mixture was refluxed overnight. After concentration, the crude product was dissolved in EtOAc, washed with water, and purified by silica gel chromoatography (EtOAc/Hex) to afford the desired furan amino ester
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