WormBase Tree Display for Construct: WBCnstr00020708
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| WBCnstr00020708 | Summary | [dpy-27promoter: mVenus-MAP::dpy-27(+): dpy-27 3'UTR] | |
|---|---|---|---|
| Driven_by_gene | WBGene00001086 | ||
| Gene | WBGene00001086 | ||
| UTR_3 | WBGene00001086 | ||
| Fusion_reporter | Venus | ||
| Type_of_construct | Nterminal_translational_fusion | ||
| Construction_summary | MAP-tagged condensin subunits were constructed using the Multisite Gateway Three-fragment Construction Kit (Invitrogen,Carlsbad, CA). For each gene, its promoter (1 kb upstream of start) and 3'UTR (0.5 kb downstream of stop) were PCR amplified with primers containing appropriate Gateway att sites and recombined into pDONRP4-P1R and pDONRP2R-P3, respectively. To create Nterminal fusion protein ORFs, the MAP tag (minus stop codon) was PCR amplified with a primer containing a 24 nt overlap with the 5' end of the gene to be tagged, and the genomic locus was amplified with a 24 nt overlap to the end of the MAP tag. These products were annealed and PCR extended with the overlap serving as primers,the full length product purified (Qiagen, Valencia, CA), then used in a final PCR amplification with outside primers containing Gateway sites and recombined into pDONR221. The three resulting entry vectors were recombined into the MosSCI destination vector,pCFJ150 (8) (Addgene, Cambridge, MA). | ||
| Used_for | Transgene_construct | WBTransgene00021109 | |
| Reference | WBPaper00047083 |
