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{"project":"craft-ca-core-dev","denotations":[{"id":"T5782","span":{"begin":83,"end":85},"obj":"PR:000006300"},{"id":"T5783","span":{"begin":92,"end":101},"obj":"SO:0000902"},{"id":"T5784","span":{"begin":115,"end":127},"obj":"CHEBI:27902"},{"id":"T5785","span":{"begin":139,"end":143},"obj":"NCBITaxon:10359"},{"id":"T5786","span":{"begin":147,"end":155},"obj":"SO:0000167"},{"id":"T5787","span":{"begin":175,"end":187},"obj":"CHEBI:27902"},{"id":"T5788","span":{"begin":227,"end":231},"obj":"NCBITaxon:10359"},{"id":"T5789","span":{"begin":232,"end":240},"obj":"SO:0000167"},{"id":"T5790","span":{"begin":300,"end":306},"obj":"SO:0000188"},{"id":"T5791","span":{"begin":338,"end":345},"obj":"SO:0000673"},{"id":"T5792","span":{"begin":338,"end":345},"obj":"CHEBI:33695"},{"id":"T5793","span":{"begin":360,"end":370},"obj":"GO:0010467"},{"id":"T5794","span":{"begin":388,"end":397},"obj":"SO:0000993"},{"id":"T5795","span":{"begin":414,"end":430},"obj":"SO:0000318"},{"id":"T5796","span":{"begin":443,"end":454},"obj":"GO:0006412"},{"id":"T5797","span":{"begin":465,"end":470},"obj":"NCBITaxon:9606"},{"id":"T5798","span":{"begin":471,"end":473},"obj":"PR:000006300"},{"id":"T5799","span":{"begin":548,"end":553},"obj":"PR:000000084"},{"id":"T5800","span":{"begin":622,"end":626},"obj":"NCBITaxon:10633"},{"id":"T5801","span":{"begin":632,"end":647},"obj":"GO:0043631"},{"id":"T5802","span":{"begin":632,"end":656},"obj":"SO:0000610"},{"id":"T5803","span":{"begin":698,"end":709},"obj":"GO:0006412"}],"text":"Standard molecular techniques were used to assemble the transgenic construct, pTRE-E2. This transgene contains the tetracycline responsive hCMV*-1 promoter [consisting of the tetracycline responsive element (TRE) and a minimal hCMV promoter] [25] from pTRE2 (Clontech Inc., Mt. View, CA), a chimeric intron from pCI (Promega) to increase message stability and expression [26,27], a Kozak consensus sequence at the initiation codon to optimize translation [28], the human E2 cDNA which has been modified to contain a 4× alanine linker followed by a c-myc epitope tag at the carboxy terminus to facilitate detection, and an SV40 late polyadenylation sequence from pCI for enhanced mRNA stability and translation."}