WormBase Tree Display for Construct: WBCnstr00020358
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WBCnstr00020358 | Other_name | [UNC-70(TSMOD)] |
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Summary | [Punc-70::UNC-70(1-1166)::TSMod::UNC-70(1167-2267)::unc-54 3'UTR] | |
Driven_by_gene | WBGene00006803 | |
Gene | WBGene00006803 | |
Construction_summary | The tension sensor, TSMod5, was inserted into UNC-70 between spectrin repeats 8 and 9, following residue 1166. This was accomplished with a four-way Gateway recombination strategy. The four entry clones contained (1) unc-70 promoter, (2) the N-terminal fragment of unc-70 (encoding residues 1-1166), (3) TSMod and (4) the carboxy-terminal fragment of unc-70(encoding residues 1167-2267) fused to the unc-54 3'UTR. The fragments were amplified by PCR using primers 1-8 (Supplementary Table 2) and combined with the appropriate DONR vector in a standard BP reaction, yielding four Entry clones:pENTR Punc-70, pENTR ATG-R8, pENTR TSMod and pENTRR9-TAA. These clones were combined in a standard LR reaction overnight with pDEST14 (Invitrogen), yielding the assembled construct Punc-70::UNC-70(1-1166)::TSMod::UNC-70(1167-2267)::unc-54 3'UTR, which was verified by sequencing. | |
Reference | WBPaper00044891 |