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{"project":"craft-ca-core-ex-dev","denotations":[{"id":"T4391","span":{"begin":1106,"end":1109},"obj":"CHEBI_SO_EXT:DNA"},{"id":"T4390","span":{"begin":1102,"end":1105},"obj":"SO_EXT:0000153"},{"id":"T4389","span":{"begin":1075,"end":1084},"obj":"SO_EXT:biological_sequence"},{"id":"T4388","span":{"begin":1068,"end":1074},"obj":"SO_EXT:0000204"},{"id":"T4387","span":{"begin":1053,"end":1057},"obj":"SO_EXT:0000147"},{"id":"T4386","span":{"begin":1041,"end":1049},"obj":"SO_EXT:biological_sequence"},{"id":"T4385","span":{"begin":999,"end":1015},"obj":"SO_EXT:coding_sequence"},{"id":"T4384","span":{"begin":972,"end":980},"obj":"SO_EXT:biological_sequence"},{"id":"T4383","span":{"begin":946,"end":960},"obj":"SO_EXT:0000132"},{"id":"T4382","span":{"begin":931,"end":939},"obj":"SO_EXT:biological_sequence"},{"id":"T4381","span":{"begin":894,"end":908},"obj":"SO_EXT:0000121"},{"id":"T4380","span":{"begin":855,"end":863},"obj":"SO_EXT:biological_sequence"},{"id":"T4379","span":{"begin":791,"end":798},"obj":"CHEBI:2511"},{"id":"T4378","span":{"begin":751,"end":763},"obj":"SO_EXT:0000006"},{"id":"T4377","span":{"begin":708,"end":717},"obj":"GO:0097617"},{"id":"T4376","span":{"begin":687,"end":699},"obj":"SO_EXT:0000006"},{"id":"T4375","span":{"begin":661,"end":665},"obj":"SO_EXT:0000704"},{"id":"T4374","span":{"begin":655,"end":660},"obj":"NCBITaxon:10088"},{"id":"T4373","span":{"begin":595,"end":609},"obj":"SO_EXT:0000121"},{"id":"T4372","span":{"begin":589,"end":594},"obj":"NCBITaxon:9606"},{"id":"T4371","span":{"begin":567,"end":575},"obj":"SO_EXT:biological_sequence"},{"id":"T4370","span":{"begin":563,"end":566},"obj":"SO_EXT:0000345"},{"id":"T4369","span":{"begin":557,"end":562},"obj":"NCBITaxon:10088"},{"id":"T4368","span":{"begin":533,"end":547},"obj":"SO_EXT:0000132"},{"id":"T4367","span":{"begin":471,"end":480},"obj":"GO:0097617"},{"id":"T4366","span":{"begin":454,"end":458},"obj":"SO_EXT:cDNA"},{"id":"T4365","span":{"begin":448,"end":453},"obj":"NCBITaxon:10088"},{"id":"T4364","span":{"begin":434,"end":442},"obj":"SO_EXT:biological_sequence"},{"id":"T4363","span":{"begin":424,"end":433},"obj":"SO_EXT:0000853"},{"id":"T4362","span":{"begin":392,"end":398},"obj":"CHEBI_SO_EXT:molecular_indicator_or_label_or_marker_or_tag"},{"id":"T4361","span":{"begin":388,"end":391},"obj":"SO_EXT:0000345"},{"id":"T4360","span":{"begin":382,"end":387},"obj":"NCBITaxon:10088"},{"id":"T4359","span":{"begin":358,"end":372},"obj":"SO_EXT:0000132"},{"id":"T4358","span":{"begin":352,"end":357},"obj":"NCBITaxon:10088"},{"id":"T4357","span":{"begin":333,"end":344},"obj":"SO_EXT:0000318"},{"id":"T4356","span":{"begin":312,"end":325},"obj":"SO_EXT:coding_sequence"},{"id":"T4355","span":{"begin":307,"end":311},"obj":"SO_EXT:cDNA"},{"id":"T4354","span":{"begin":299,"end":303},"obj":"PR_EXT:ACDP_or_CNNM"},{"id":"T4353","span":{"begin":293,"end":298},"obj":"NCBITaxon:9606"},{"id":"T4352","span":{"begin":267,"end":281},"obj":"SO_EXT:0000121"},{"id":"T4351","span":{"begin":252,"end":256},"obj":"PR_EXT:ACDP_or_CNNM"},{"id":"T4350","span":{"begin":222,"end":229},"obj":"CHEBI_SO_EXT:molecular_indicator_or_label_or_marker_or_tag"},{"id":"T4349","span":{"begin":218,"end":221},"obj":"SO_EXT:0000345"},{"id":"T4348","span":{"begin":201,"end":204},"obj":"SO_EXT:0000345"},{"id":"T4347","span":{"begin":195,"end":200},"obj":"NCBITaxon:10088"},{"id":"T4346","span":{"begin":161,"end":170},"obj":"SO_EXT:biological_sequence"},{"id":"T4345","span":{"begin":158,"end":160},"obj":"CHEBI_SO_EXT:amino_acid"},{"id":"T4344","span":{"begin":138,"end":143},"obj":"SO_EXT:cDNA"},{"id":"T4343","span":{"begin":133,"end":137},"obj":"PR_EXT:ACDP_or_CNNM"},{"id":"T4342","span":{"begin":127,"end":132},"obj":"NCBITaxon:9606"},{"id":"T4341","span":{"begin":76,"end":85},"obj":"SO_EXT:biological_sequence"},{"id":"T4340","span":{"begin":66,"end":75},"obj":"SO_EXT:0000853"},{"id":"T4339","span":{"begin":60,"end":65},"obj":"NCBITaxon:9606"},{"id":"T4338","span":{"begin":25,"end":29},"obj":"SO_EXT:0000704"},{"id":"T4337","span":{"begin":20,"end":24},"obj":"PR_EXT:ACDP_or_CNNM"},{"id":"T4336","span":{"begin":5,"end":12},"obj":"SO_EXT:sequence_cloning_process"},{"id":"T4335","span":{"begin":0,"end":4},"obj":"SO_EXT:cDNA"}],"text":"cDNA cloning of the Acdp gene family was performed based on human homologue sequences as previously reported [2]. Briefly, the human ACDP cDNAs and predicted AA sequences were used to search the mouse EST database for EST markers corresponding to each Acdp member. A forward primer within the human ACDP 5' cDNA coding region (after start codon) and a mouse reverse primer from the mouse EST marker were used to amplify the homologue sequence from mouse cDNA at very low annealing temperature (45–50°C). A nested PCR using an inside reverse primer from the mouse EST sequence and the same human forward primer was then carried out to amplify the specific mouse gene from the first round PCR products at high annealing temperature (62°C). The expected PCR products were directly excised from agarose gel and sequenced by an ABI377 automatic sequencer. The sequence was further confirmed using a forward primer from newly identified sequence and a reverse primer from known sequence. Once most of the coding sequences were identified, partial sequence of exon 1 and the 5' UTR sequences were obtained by BAC DNA sequencing."}