Flavonoid <i>C</i>-glucosides, which are found in several plant families, are characterized by several biological properties, including antioxidant, anticancer, anti-inflammatory, neuroprotective, hepatoprotective, cardioprotective, antibacterial, antihyperalgesic, antiviral, and antinociceptive activities. The biosynthetic pathway of flavonoid <i>C</i>-glucosides in plants has been elucidated. In the present study, a pathway was introduced to <i>Escherichia coli</i> to synthesize four flavonoid <i>C</i>-glucosides, namely, isovitexin, vitexin, kaempferol 6-<i>C</i>-glucoside, and kaempferol 8-<i>C</i>-glucoside. A five- or six-step metabolic pathway for synthesizing flavonoid aglycones from tyrosine was constructed and two regioselective flavonoid <i>C</i>-glycosyltransferases from <i>Wasabia japonica</i> (<i>WjGT1</i>) and <i>Trollius chinensis</i> (<i>TcCGT</i>) were used. Additionally, the best shikimate gene module construct was selected to maximize the titer of each <i>C-</i>glucoside flavonoid. Isovitexin (30.2 mg/L), vitexin (93.9 mg/L), kaempferol 6-<i>C</i>-glucoside (14.4 mg/L), and kaempferol 8-<i>C</i>-glucoside (38.6 mg/L) were synthesized using these approaches. The flavonoid <i>C</i>-glucosides synthesized in this study provide a basis for investigating and unraveling their novel biological properties.