Detailed protocols

Single-copy insertion

Constitutive Cas9 (plasmid-based)

Heat shock Cas9 (plasmid-based)

Multiplexed w heat shock Cas9 (plasmid-based)

Extra-chromosomal array integration

Two-step Cas9 (protein + sgRNA).

One-step constitutive Cas9 (plasmid).

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Reagents

Strains

Site	Selection	Strain	Annotated genomic regions (GenBank “right click and save” – please ignore browser warning)	CGC
Chr. I (7.45 Mb)	unc-119	CFJ77	1. Genomic 2. Landing site (no insert) 3. Landing site (w insert)	Strain request
	Pmlc-2::gfp	CFJ184	1. Genomic 4. Landing site (no insert) 5. Landing site (w insert)	Strain request
Chr. II (8.24 Mb)	unc-119	CFJ42	1. Genomic 2. Landing site (no insert) 3. Landing site (w insert)	Strain request
	Pmlc-2::gfp	CFJ108	1. Genomic 4. Landing site (no insert) 5. Landing site (w insert)	Strain request
	HygroR	CFJ111	1. Genomic 6. Landing site (no insert) 7. Landing site (w insert)	Strain request
Chr. IV (8.62 Mb)	unc-119	CFJ94	1. Genomic 2. Landing site (no insert) 3. Landing site (w insert)	Strain request