Figure 1
From: Generation of stable Drosophila cell lines using multicistronic vectors
Schematic representation of the p Ac5-STABLE vectors.
All constructs were based on the vector backbone of pAc5.1 (Invitrogen), with expression driven by the Actin5C promoter. (A) In pAc5-GFP, GFP alone served as a negative control for selection experiments. (B) In pAc5-STABLE1-Neo, designed for bicistronic expression, GFP and NeoR are separated by the T2A sequence. Unique sites are shown. (C) pAc5-STABLE2-Neo, designed for tricistronic expression, features FLAG-tagged mCherry, GFP and NeoR, each separated by a T2A peptide. In the case of dT2A, it has identical peptide sequence to T2A, but degenerated sequence at the nucleotide level. Unique sites are shown. (D) pAc5-dTES.GFP-Neo was constructed by cloning Drosophila CG6522 into the EcoRI-XbaI sites of pAc5-STABLE1-Neo, generating an N-terminal fusion to GFP.
