REAGENT or RESOURCE	SOURCE	IDENTIFIER
Bacterial and virus strains
Escherichia coli Dh5α	Thermo Fisher Scientific	N/A
Chemicals, peptides, and recombinant proteins
Uridine	Merck KGaA	Cat#U3003
L-Leucine	Merck KGaA	Cat#L8000
5 Fluoroorotic acid monohydrate (5-FOA)	Apollo Scientific	Cat#PC4054
EDTA	Merck KGaA	Cat#EDS
Tris	Merck KGaA	Cat#T1503
Niacine (nicotinc acid)	Merck KGaA	Cat#N0761
Thiamine	Merck KGaA	Cat#T1270
D-Sorbitol	Merck KGaA	Cat#S1876
Lysing Enzymes	Merck KGaA	Cat#L412
Chitosanase	Merck KGaA	Cat#C9830
DMSO	Merck KGaA	Cat#D8418
SDS	Merck KGaA	Cat#L3771
RNase A	Merck KGaA	Cat#R6513
Phenol	Merck KGaA	Cat#P1037
Chloroform	Merck KGaA	Cat#C2432
Isoamyl alcohol	Merck KGaA	Cat#309435
Ammonium acetate	Merck KGaA	Cat#A1542
Ethanol	Merck KGaA	Cat#51976
Hydrochloric acid	Merck KGaA	Cat#H1758
Critical commercial assays
Alt-R™ CRISPR-Cas9 crRNA	IDT	See Table S1
Alt-R™ CRISPR-Cas9 tracrRNA	IDT	Cat#1072533
Alt-R™ S.p. Cas9 Nuclease V3	IDT	Cat#1081058
Deposited data
Mucoromycotina genomes	Joint Genome Institute Mycocosm	https://mycocosm.jgi.doe.gov/mucoromycotina/mucoromycotina. info.html
R. microsporus pyrG	Joint Genome Institute Mycocosm	R. microsporus Protein ID 279857
R. microsporus pyrF	Joint Genome Institute Mycocosm	R. microsporus Protein ID 231940
R. microsporus leuA	Joint Genome Institute Mycocosm	R. microsporus Protein ID 228594
R. microsporus crgA	Joint Genome Institute Mycocosm	R. microsporus Protein ID 226533
Experimental models: Organisms/strains
Rhizopus microsporus ATCC 11559	Kindly provided by Dr. Teresa E. Pawlowska (Cornell University)	N/A
Rhizopus microsporus UM1: pyrF-	This Study	N/A
Rhizopus microsporus UM3: crgA::pyrF	This Study	N/A
Rhizopus microsporus UM4: crgA::pyrF	This Study	N/A
Rhizopus microsporus UM6: leuA::pyrF	This Study	N/A
Rhizopus microsporus UM7: leuA::pyrF	This Study	N/A
Mucor lusitanicus CBS 277.49	von Arx and Schipper, 1978	N/A
Mucor lusitanicus MU223: crgAΔ::pyrG	Silva et al., 2008	N/A
Oligonucleotides
Oligonucleotides	This Study	See Table S1
Recombinant DNA
pBluescript II SK+	Agilent	Cat#212205
pMAT1819	This Study	N/A
Software and algorithms
Snapgene v5.1.0	Snapgene Software	https://www.snapgene.com/
EuPaGDT	Peng and Tarleton, 2015	http://grna.ctegd.uga.edu/
NIS Elements Software	Nikon	https://www.microscope.healthcare.nikon.com/products/software/nis-elements
ImageJ v1.51m9	Schneider et al., 2012
Zeiss Zen v2.5	Zeiss	https://www.zeiss.com/microscopy/us/products/microscope-software/zen.html#introduction
Jmol		http://jmol.sourceforge.net/
Mol∗		http://molstar.org
GraphPad Prism v8.0.2	Graphpad Software	https://www.graphpad.com/
IBM SPSS Statistic	IBM Software	https://www.ibm.com/analytics/spss-statistics-software