The invention discloses a CD123 single domain antibody, a nucleic acid sequence, an expression vector and a kit. An amino acid sequence of the CD123 single domain antibody includes an FR1 region, a CDR1 region, an FR2 region, a CDR2 region, an FR3 region, a CDR3 region and an FR4 region which are formed in sequence, wherein the FR1 region, the FR2 region, the FR3 region and the FR4 region are composed of frame-region amino acid sequences, the CDR1 region, the CDR2 region and the CDR3 region are composed of variable-region amino acid sequences, and the amino acid sequences of the FR1 region, the CDR1 region, the FR2 region, the CDR2 region, the FR3 region, the CDR3 region and the FR4 region are shown in a table 1. According to the technical scheme, in combination with a genetic engineeringmethod, the single domain antibody capable of specifically binding to a CD123 protein is obtained, and the single domain antibody has high specific binding capability of specifically binding to the CD123 protein and high affinity.