Speckled Protein 140 (SP140) is an immune-specific chromatin reader belonging to the Speckled Protein family including SP100, SP110, and SP140L. Although expressed by different immune cell types, SP140 is highly expressed in B cells and mutations in SP140 have been linked to B cell–mediated diseases but how it targets and occupies the genome remains poorly understood. To investigate this, we mapped genome-wide SP140 binding sites in human B cells with CUT&Tag. In contrast to previous reports that SP140 primarily binds repressed chromatin, we found that SP140 binds ubiquitously to non-methylated CpG-rich promoters marked by active (H3K4me3) and repressive (H3K27me3) histone modifications. To determine whether SP140 specifically targets non-methylated DNA, we confirmed that the SP140 SAND can specifically bind to CpG-containing DNA via its KNWR motif, and its ability to bind DNA in vitro was reduced by DNA methylation. To explore if DNA methylation blocks SP140 occupancy in cells, we treated B cells with decitabine to inhibit DNA methylation. We identified increased SP140 occupancy at previously hyper-methylated regions, consistent with novel targeting in the absence of DNA methylation at non-methylated gene promoters. To subsequently investigate whether SP140 is required for normal transcriptional activity at its target gene promoters, we knocked in an FKBP-V degron tag into the endogenous SP140 locus in a B cell line. We found that SP140 loss did not result in major changes in gene expression at steady state or following IFNa treatment, contradicting reports that SP140 is required for transcriptional activation of IFN-stimulated genes. Similarly, we did not detect gene expression changes after CRISPR/Cas9-mediated deletion of the 400kb Speckled Protein family region. These results demonstrate that SP140 is not required for maintenance of normal gene expression in this cell line but may be important for other context-specific transcriptional responses through its occupancy of non-methylated gene promoters genome-wide.