Comments: Combinatorial libraries of zinc finger arrays made by OPEN are cloned into this plasmid (see Maeder et al., Mol Cel 2008, PMID 18657511). Ligate zinc finger array fragments with BbsI-digested pBR-UV5-GP-FD2 vector backbone to create plasmids that express the zinc finger array as a FLAG-tagged Gal11P fusion in the B2H system.

The lacUV5 promoter which drives expression of the Gal11P fusion protein contains a lac operator and therefore can be repressed by lac repressor.

This plasmid is a low copy number plasmid and therefore we recommend that minipreps be performed from a 4 to 10 ml culture.

Annotations for pMLM36:

12-17: lacUV5 promoter -35 box / 36-41: lacUV5 promoter -10 box / 48: lacUV5 promoter transcription start point / 86-88: Start codon for Gal11P protein fragment / 89-358: Coding sequence for Gal11P a.a. 263-352 (with additional translationally silent mutations) / 407-412: BbsI site #1 / 433-438: BbsI site #2 / 568-1024: f1 phage origin of replication / 1156-2013: beta-lactamase gene cassette (confers resistance to ampicillin and carbenicillin) / 2016-2956: ColE1 origin of replication (plasmid origin) / 3203-3390: rop gene (associated with regulation of copy number)

Addgene has sequenced a portion of this plasmid for verification. Click here for the sequencing result.