The antimalarial drug chloroquine is the first line of drug treatment for P. vivax in most countries and has proven highly efficacious. However, chloroquine-resistant (CQR) P. vivax infections have been widely reported. The molecular determinants of CQR and resistance to other antimalarials in P. vivax remain unclear, largely due to the lack of in vitro culture, precluding reverse genetics. Several candidate drug transporter genes have been identified in P. vivax with polymorphism in sequence and expression level that could be associated with CQR and/or resistance to other antimalarials, including pvmdr1, orthologous to Plasmodium falciparum pfmdr1 a key determinant of antimalarial susceptibility in P. falciparum. Despite years of research focusing on pvmdr1, no definitive relationship between pvmdr1 polymorphisms and drug resistance has emerged. P. knowlesi, a zoonotic macaque parasite closely related to P. vivax, can be cultured and has been used as a model system to characterize P. vivax genes. We first analyzed a P. vivax population genomic dataset to identify a set of 24 geographically representative circulating pvmdr1 haplotypes alleles. These pvmdr1 alleles were introduced into P. knowlesi in place of the endogenous pkmdr1 gene, and transgenic lines were assayed for resistance to an array of antimalarial compounds. We found that certain haplotypes confer resistance (2- to 4-fold) to mefloquine, dihydroartemisinin, and lumefantrine. Other haplotypes conferred sensitivity. However, we find no evidence that pvmdr1 haplotypes alone confer a significant shift in chloroquine susceptibility. Our results suggest that it is the co-selection of pvmdr1 by other antimalarials used to treat P. falciparum that has selected for certain pvmdr1 haplotypes. Our results have implications for guiding P. vivax treatment policy and molecular surveillance of P. vivax drug resistance alleles.