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pBS, then mutated the HindIII site into a FseI site. We next mutated the C-terminal part of the p53 gene in two rounds of PCR mutagenesis, first with primers 5′-GGGCCTGACTCAGACGGATCCCCTCTGCATCCCGTC-3′ and 5′-GACGGGATGCAGAGGGGATCCGTCTGAGTCAGGCCC-3′, which removed the stop codon and introduced a BamHI site, then with primers 5′-GACGGATCCCCTCTGAATTCCGTCCCCATCACCA-3′ and 5′-TGGTGATGGGGACGGAATACAGAGGGGATCCGTC-3′, which introduced an EcoRI site. We verified the sequence from the mutated plasmid, then digested it with BamHI and EcoRI, to insert in frame GFP sequences from a Bam HI-EcoRI fragment of plasmid phr-GFP-1 (Stratagene). We verified the sequence of this p53-GFP fusion fragment, then swapped it in the L3-p53PmlEagPuroΔTK-1L plasmid (see above) by HindIII and FseI digestion, resulting "}

{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T5711","span":{"begin":14,"end":21},"obj":"SO_EXT:sequence_alteration_process"},{"id":"T5712","span":{"begin":26,"end":33},"obj":"PR_EXT:P43870"},{"id":"T5713","span":{"begin":65,"end":72},"obj":"SO_EXT:sequence_alteration_process"},{"id":"T5714","span":{"begin":77,"end":87},"obj":"CHEBI_SO_EXT:C_terminus_or_C_terminal_region"},{"id":"T5715","span":{"begin":88,"end":95},"obj":"_FRAGMENT"},{"id":"T5716","span":{"begin":104,"end":108},"obj":"SO_EXT:0000831"},{"id":"T5717","span":{"begin":100,"end":103},"obj":"PR_EXT:000003035"},{"id":"T5718","span":{"begin":130,"end":141},"obj":"SO_EXT:sequence_alteration_process"},{"id":"T5719","span":{"begin":154,"end":161},"obj":"SO_EXT:0000112"},{"id":"T5720","span":{"begin":271,"end":281},"obj":"SO_EXT:0000319"},{"id":"T5721","span":{"begin":299,"end":304},"obj":"PR_EXT:P23940"},{"id":"T5722","span":{"begin":321,"end":328},"obj":"SO_EXT:0000112"},{"id":"T5723","span":{"begin":464,"end":472},"obj":"SO_EXT:biological_sequence"},{"id":"T5724","span":{"begin":482,"end":489},"obj":"SO_EXT:sequence_alteration_process"},{"id":"T5725","span":{"begin":490,"end":497},"obj":"SO_EXT:0000155"},{"id":"T5726","span":{"begin":521,"end":526},"obj":"PR_EXT:P23940"},{"id":"T5727","span":{"begin":541,"end":547},"obj":"SO_EXT:sequence_insertion_process"},{"id":"T5728","span":{"begin":548,"end":556},"obj":"SO:0001817"},{"id":"T5729","span":{"begin":561,"end":570},"obj":"SO_EXT:biological_sequence"},{"id":"T5730","span":{"begin":578,"end":584},"obj":"PR_EXT:P23940"},{"id":"T5731","span":{"begin":603,"end":610},"obj":"SO_EXT:0000155"},{"id":"T5732","span":{"begin":651,"end":659},"obj":"SO_EXT:biological_sequence"},{"id":"T5733","span":{"begin":668,"end":671},"obj":"PR_EXT:000003035"},{"id":"T5734","span":{"begin":676,"end":682},"obj":"SO_EXT:sequence_fusion_entity_or_process"},{"id":"T5735","span":{"begin":739,"end":746},"obj":"SO_EXT:0000155"},{"id":"T5736","span":{"begin":762,"end":769},"obj":"PR_EXT:P43870"}],"relations":[{"id":"R1617","pred":"_lexicallyChainedTo","subj":"T5716","obj":"T5715"}],"text":"nto pBS, then mutated the HindIII site into a FseI site. We next mutated the C-terminal part of the p53 gene in two rounds of PCR mutagenesis, first with primers 5′-GGGCCTGACTCAGACGGATCCCCTCTGCATCCCGTC-3′ and 5′-GACGGGATGCAGAGGGGATCCGTCTGAGTCAGGCCC-3′, which removed the stop codon and introduced a BamHI site, then with primers 5′-GACGGATCCCCTCTGAATTCCGTCCCCATCACCA-3′ and 5′-TGGTGATGGGGACGGAATACAGAGGGGATCCGTC-3′, which introduced an EcoRI site. We verified the sequence from the mutated plasmid, then digested it with BamHI and EcoRI, to insert in frame GFP sequences from a Bam HI-EcoRI fragment of plasmid phr-GFP-1 (Stratagene). We verified the sequence of this p53-GFP fusion fragment, then swapped it in the L3-p53PmlEagPuroΔTK-1L plasmid (see above) by HindIII and FseI digestion, resulting "}

{"project":"craft-ca-core-dev","denotations":[{"id":"T5676","span":{"begin":26,"end":33},"obj":"PR:P43870"},{"id":"T5677","span":{"begin":88,"end":95},"obj":"_FRAGMENT"},{"id":"T5678","span":{"begin":104,"end":108},"obj":"SO:0000831"},{"id":"T5679","span":{"begin":100,"end":103},"obj":"PR:000003035"},{"id":"T5680","span":{"begin":154,"end":161},"obj":"SO:0000112"},{"id":"T5681","span":{"begin":271,"end":281},"obj":"SO:0000319"},{"id":"T5682","span":{"begin":299,"end":304},"obj":"PR:P23940"},{"id":"T5683","span":{"begin":321,"end":328},"obj":"SO:0000112"},{"id":"T5684","span":{"begin":490,"end":497},"obj":"SO:0000155"},{"id":"T5685","span":{"begin":521,"end":526},"obj":"PR:P23940"},{"id":"T5686","span":{"begin":548,"end":556},"obj":"SO:0001817"},{"id":"T5687","span":{"begin":578,"end":584},"obj":"PR:P23940"},{"id":"T5688","span":{"begin":603,"end":610},"obj":"SO:0000155"},{"id":"T5689","span":{"begin":668,"end":671},"obj":"PR:000003035"},{"id":"T5690","span":{"begin":739,"end":746},"obj":"SO:0000155"},{"id":"T5691","span":{"begin":762,"end":769},"obj":"PR:P43870"}],"relations":[{"id":"R1616","pred":"_lexicallyChainedTo","subj":"T5678","obj":"T5677"}],"text":"nto pBS, then mutated the HindIII site into a FseI site. We next mutated the C-terminal part of the p53 gene in two rounds of PCR mutagenesis, first with primers 5′-GGGCCTGACTCAGACGGATCCCCTCTGCATCCCGTC-3′ and 5′-GACGGGATGCAGAGGGGATCCGTCTGAGTCAGGCCC-3′, which removed the stop codon and introduced a BamHI site, then with primers 5′-GACGGATCCCCTCTGAATTCCGTCCCCATCACCA-3′ and 5′-TGGTGATGGGGACGGAATACAGAGGGGATCCGTC-3′, which introduced an EcoRI site. We verified the sequence from the mutated plasmid, then digested it with BamHI and EcoRI, to insert in frame GFP sequences from a Bam HI-EcoRI fragment of plasmid phr-GFP-1 (Stratagene). We verified the sequence of this p53-GFP fusion fragment, then swapped it in the L3-p53PmlEagPuroΔTK-1L plasmid (see above) by HindIII and FseI digestion, resulting "}