pAmCyan載體描述pAmCyan is a pUC19-derived prokaryotic expression vector, which encodes a variant of wildtype Anemonia majano cyan fluorescent protein (AmCyan; 1) that has been engineered for brighter fluorescence. Two amino acid substitutions (Asn-34 to Ser; Lys-68 to Met) have been made to enhance the emission characteristics of AmCyan (excitation maximum = 458 nm;emission maximum = 489 nm).The AmCyan gene was inserted in frame with the lacZ initiation codon from pUC19 so that AmCyan is expressed from the lac promoter (Plac) in E. coli. The AmCyan coding sequence is flanked by distinct multiple cloning sites (MCS) at the 5' and 3' ends so that the gene can be readily excised from pAmCyan and subcloned into other expression vectors. An upstream sequence—located just 5' to the AmCyan gene—has been converted to a Kozak consensus translation initiation site (2) to increase the translation efficiency in eukaryotic expression systems. The pUC backbone of pAmCyan provides a high-copy-number origin of replication (pUC ori) and an ampicillin resistance gene (Ampr) for propagation and selection in E. coli.pAmCyan Vector serves as a convenient source of AmCyan cDNA. The flanking MCS regions make it possible to excise the AmCyan coding sequence and insert it into other expression vectors. Alternatively, the AmCyan coding sequence can be amplified by PCR. Propagation in E. coli Recommended host strain: JM109 Selectable marker: plasmid confers resistance to ampicillin (50 μg/ml) to E. coli hosts E. coli replication origin: pUC Copy number: ~500 Plasmid incompatibility group: pMB1/Col E1