Construction notes
From Chang et al., 2021:
Construction of pWALEXA20 (White-AttB-LexAop) vector
To replace 10xUAS in the pWALIUM20 vector with 13xLexAop2 (sulA derived LexA DNA-binding
motifs), a 675 bp product was amplified from pJFRC19-13xLexAop2-IVS-CD4-myr::GFP (Pfeiffer et al
2010) using primers LexAop_F1 (5’-CACCCATGCATAGGGCCGCAAGCTTGCATG-3’) and
hsp70p_R (5’-CCTTTTAGATCTATTCAGAGTTCTCTTCTTGTATTCAATAATTACTTCTTGGC-3’).
The PCR product was inserted to the PstI and BglII sites on the pWALIUM20 vector (Perkins 2015) by
Gibson-assembly (NEB HiFi DNA Assembly Kit) to generate pWALEXA20 (White-AttB-LexAop for
shRNA expression), a phiC31 integrase-mediated transformation vector with mini-white as a selection marker.