pPV32

2 kb of the gpa-4 promoter was inserted upstream of human caspase in the pV32 plasmid (a kind gift from V. Maricq) using PstI and KpnI restriction sites.

IM#182

IM#97 is the plasmid construct of unc-6 genomic sequence. The plasmid pIM#182 was made by deleting domain V-3. PIM#97 was partially digested with XbaI, a 12375 bp fragment was eluted from the digestion mixture, and this fragment was self ligated to generate pIM#182. --precise ends.

pCMH1195

The PEST domain was amplified from pAF207 (a generous gift from Dr. A. Frand, UCLA) and was used to create NLS-mCherry-pest::unc-54-3'UTR (pCMH1183). To make pCMH1195, a 2kb genomic fragment containing the col-12 promoter (chr V:10,422,005-10,429,220) was subcloned into the XmaI sites of pCMH1183.

IM#181

IM#97 is the plasmid construct of unc-6 genomic sequence. The plasmid pIM#181 was constructed by deleting domain V-2. The pIM#97 was digested with XhoI and subjected to a Klenow reaction without dNTP. The fragments were separated by agarose gel electrophoresis, and the largest fragment was eluted, digested with NruI, and self ligated to generate pIM#181. --precise ends.

pDNL10

N2 genomic lysates were PCR amplified with primers B4F2 and B1R3 (see Table S1 for primer sequences). The 5.6 kb fragment was recombined into pDONR P4P1R. This construct has 1169 bp of 5' UTR sequence. Sequencing revealed an error at nucleotide #1921 (T->C nucleotide change; V->A amino acid change), which was corrected with the QuikChange Mutagenesis kit (Agilent Technologies, Santa Clara, CA). Theentry clone containing the cbd-1 3' UTR was made as follows. N2 genomic lysates were PCR amplified with primers B2rF2 and B3R1. The 389 bp PCR fragment was recombined into pDONR P2RP3. The expression clone was constructed by performing a Gateway LR reaction with the two entryclones, pCR347, and pCR319.

IM#206

IM#97 is the plasmid construct of unc-6 genomic sequence. The plasmid pIM#206 was constructed by deleting domain V-1. To make this deletion construct, the two fragments that introduce a SphI site were amplified by PCR with pIM#97 as a template. For the first fragment, 5 -GAGAACAGACCTTCTGC-3 and 5 -GATCGCATGCACCACCAACTGCC-3 were used as a set of primers. This PCR product was double digested with SgrAI and SphI. For the second fragment, 5 -GATCGCATGCGCTTGCAACTGCA-3 and 5 -CACTACAAAAAGGCGCCTGG-3 were used as primers for the PCR with the same template. SphI and NarI were used to digest this amplified DNA. An 1884 bp fragment was excised with SgrAI and NarI from pIM#97 and replaced by the above SgrAISphI and SphINarI fragments to generate pIM#206. --precise ends.

WBCnstr00022583

a mRFP-tagged version of Annexin V (sAnxV::mRFP) that binds PS (Mapes et al., 2012)

WBCnstr00013875

translational fusion. Full-length trim-9 cDNA was amplified from the cDNA clone yk801c07 and inserted into the delta-PSM vector between the Nhe I and EcoR V sites.

WBCnstr00004794

Mapping info: The uIs9 insertion was mapped within 1 m.u. of unc-23 in the central cluster of chromosome V [none of 42 Unc progeny from unc-23(e25)/uIs9 exhibited GFP fluorescence].

WBCnstr00012951

[f14d12.6::gfp] full-length translational fusion. Primers were used to amplify a 3987 bp fragment upstream from the start ATG and the 5' end of the f14d12.6 genomic sequence including the first 53 nt of exon V. --precise ends.