Anopheles coluzzii Enhancer Mapping

Mosquito larvae were collected in Goundry village, Burkina Faso (latitude 12.5166876, longitude −1.3921092) using described methods^{39 (link)}, reared to adults, and were typed for species by the SINE200 X6.1 assay^{40 (link)}. DNA from 60 A. coluzzii were pooled at equal volume and sheared using an S220 ultrasonicator (Covaris) to produce DNA fragments 800–1000 bp in length. Subsequently, DNA was processed as described for the STARR-seq assay^{30 (link)}, cloned into the plasmid pSTARR-seq_fly (AddGene 71499), transformed into MegaX DH10B T1R Electrocomp Cells (Invitrogen), cultured in LB+ ampicillin (1 ug/ml), and plasmid DNA was purified using the Plasmid Plus Mega Kit (Qiagen). The Anopheles gambiae PEST AgamP4 genome assembly available at Vectorbase was used as the reference genome (https://www.vectorbase.org/organisms/anopheles-gambiae/pest/agamp4).