SP-A is the most abundant, surfactant-associated protein isolated from lung lavage. Genomic blot analysis of total human cellular DNA with SP-A cDNA demonstrated the presence of multiple hybridizing fragments that are not accounted for by available SP-A gene sequences. In this report, we have cloned and characterized human genomic DNA fragments that account for some of the other hybridizing fragments. These clones contain nucleotide sequences that are highly homologous to the fourth intron and fifth exon of the human SP-A gene. Sequences upstream from these SP-A-like sequences are not detectable by Northern blot hybridization of SP-A-expressing cells and the SP-A-like sequences contain premature stop codons, consistent with the interpretation that these clones represent an SP-A pseudogene. Restriction fragments consistent with this pseudogene and the functional SP-A gene are present in a human chromosome 10 genomic library made from a single chromosome, showing that the functional SP-A gene and the pseudogene are syntenic.