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. 2008 Sep;180(1):103–119. doi: 10.1534/genetics.108.090274

TABLE 1.

FLP recombinase expression constructs and FLP-out targets

A. FLP recombinase expression constructs
Plasmid Promoter Genea unc-119(+)b
pGC94 hsp-16.2 FLP No
pGC95 hsp-16.41 FLP No
pGC133 hsp-16.41 FLP Yes
pGC146 hsp-16.2 FLP Yes
pGC158 lag-2 FLP No
B. FLP recombinase Gatewayd destination vectors to generate expression constructs in the form [promoter-FLP recombinase]
Plasmid attd attd iAc Genea unc-119(+)b
pGC180 R1 R2 None FLP Yes
pGC181 R2 R1 None FLP Yes
pGC267 R1 R2 Yes FLP Yes
C. FRT-containing full FLP-out target cassettes in the form [promoter<GFP<gene/reporter/hairpin]
Plasmid att Promoter att FRT iAc Genea unc-119(+)b FRT iAc att Gene att 3′end unc-119(+)b
pGC185 B1 rpl-28 B2 Yes No 4xNLS-GFP Yes Yes No No 4xNLS-tdimer2(12) No let-858 No
pGC200 B1 pro-1 B2 Yes No 4xNLS-GFP Yes Yes Yes No 1xNLS-lacZ No unc-54 No
pGC219 B1 hlh-12 B2 Yes Yes 4xNLS-GFP No Yes Yes B4 hlh-12(R15K) B3 let-858 Yes
pGC220 B1 hlh-12 B2 Yes Yes 4xNLS-GFP No Yes Yes B4 hlh-12(R25K) B3 let-858 Yes
pGC240 B1 lim-7 B2 Yes Yes 4xNLS-GFP No Yes Yes No 4xNLS-tdimer2(12) No let-858 Yes
pGC452 B4 hlh-12 B3 Yes Yes 4xNLS-GFP No Yes Yes B1 hairpin_hlh-12 B1 let-858 Yes
D. FRT-containing Gateway destination vectors in the form [Gatewayd (R1-R2)<reporter<reporter] for insertion of promoter upstream of FLP-out cassette containing a reporter in the cassette and a reporter downstream of the cassette
Plasmid attd attd FRT iA Genea unc-119(+)b FRT iA att Gene att 3′end unc-119(+)b
pGC93 R1 R2 Yes No 4xNLS-GFP Yes Yes No No 1xNLS-lacZ No unc-54 No
pGC97 R1 R2 Yes No 4xNLS-GFP Yes Yes No No tdimer2(12) No let-858 No
pGC183 R1 R2 Yes No 4xNLS-GFP Yes Yes No No 4xNLS-tdimer2(12) No let-858 No
pGC238 R1 R2 Yes Yes 4xNLS-GFP No Yes Yes No 4xNLS-tdimer2(12) No let-858 Yes
pGC160 R1 R2 Yes No tdimer2(12) No Yes No No 4xNLS-GFP No let-858 Yes
E. FRT-containing Gateway destination vectors in the form [Gatewayd (R1-R2)<reporter<Gatewayd (R4-R3)] for insertion of promoter upstream of FLP-out cassette and gene of interest downstream of FLP-out cassette
Plasmid attd attd FRT iA Genea unc-119(+)b FRT iA attd attd 3′end unc-119(+)b
pGC162 R1 R2 Yes No 4xNLS-GFP Yes Yes No R4 R3 let-858 No
pGC247 R1 R2 Yes Yes 4xNLS-GFP No Yes No R4 R3 let-858 Yes
pGC163 R1 R2 Yes No tdimer2(12) No Yes No R4 R3 let-858 Yes
pGC225 R1 R2 Yes No 4xNLS-tdimer2(12) No Yes No R4 R3 let-858 Yes
pGC276 R1 R2 Yes Yes 4xNLS-tdimer2(12) No Yes Yes R4 R3 let-858 Yes
F. FRT-containing Gateway destination vectors in the form [Gatewayd (R4-R3)<reporter<Wormgatee] for insertion of promoter upstream of FLP-out cassette and RNAi hairpin downstream of FLP-out cassette
Plasmid attd attd FRT iA Genea unc-119(+)b FRT iA att att 3′end unc-119(+)b
pGC245 R4 R3 Yes Yes 4xNLS-GFP No Yes Yes R1 Wormgatee R1 let-858 Yes
pGC320 R4 R3 Yes Yes 4xNLS-mCherry No Yes Yes R1 Wormgatee R1 let-858 No
G. Donor vector for Gateway(R4-R3) cassette
Plasmid
pGC188f

See materials and methods or supplemental Methods for plasmid construction details. See also supplemental Toolkit Documentation and supplemental table for strategies and additional constructs to facilitate construction of FLP- and FRT-containing plasmids.

a

All plasmids contain the let-858 3′end after the FLP recombinase and all reporters [GFP, tdimer2(12), or mCherry]. This sequence adds the let-858 3′-UTR onto the transcript.

b

C. briggsae unc-119(+); see text for details.

c

Intron A from Fire vectors.

d

Whole Gateway cassettes (with ccdB gene and chloramphenicol resistance gene; Invitrogen) are between attR sites unless otherwise indicated.

e

Wormgate (Johnson et al. 2005) consists of two reciprocally oriented Gateway cassettes flanking an intron. This system accepts cDNA clones directly from the ORFeome library.

f

See materials and methods and supplemental Toolkit Documentation for details on pGC188.