A simplified counter-selection recombineering protocol for creating fluorescent protein reporter constructs directly from C. elegans fosmid genomic clones

Table 1 pGOv5-based constructs ^a

Construct	Insert	Intron 1^e	Intron 2
pNH001	[(G₄S)₃]::C-TAP-tag::2xNLS ^b	-	-
pNH002	N-TAP-tag::[(G₄S)₃]::mTFP1 ^c	-	-
pNH009	N-TAP-tag::mTFP1	-	-
pNH013	mTFP1 ^d	-	-
pNH026	mTFP1::[(G₄S)₃]::C-TAP-tag	-	-
pNH030	mTFP1::[(G₄S)₃]::2xNLS	-	-
pNH078	mTFP1[1I] ^d	B	-
pNH082	mTFP1[2I] ^d	B	C

^a All constructs are in E. coli DH5α and are available from Addgene ^b lacks intact open-reading frame; ^c mTFP1 sequence lacks N-terminal Met-Val and C-terminal Lys residues present in native sequence; ^d N-terminal Met-Val residues and C-terminal Lys residue, present in native coding sequence, replaced, respectively, with Met-Ala-Ala and Val-Ser-Ala; ^e see Additional file 1: Table S2.

ISSN: 1472-6750