Et(E1B:FRT-GAL4-VP16-FRT-Cre,myl7:GFP)

ID

ZDB-ETCONSTRCT-130724-1

Name

Et(E1B:FRT-GAL4-VP16-FRT-Cre,myl7:GFP)

Previous Names

• Et(bp:FRT-Gal4VP16-FRT-Cre) (1)
• Et(E1b:FRT-GAL4-VP16-FRT-Cre,myl7:GFP)
• Et(E1b:FRT-Gal4-VP16-FRT-Cre,myl7:GFP)

Type

engineered_plasmid

Regulatory Regions

• E1B
• myl7 (1)

Coding Sequences

• Cre (1)
• GAL4 (1)
• GFP (1)
• VP16 (1)

Contains Sequences

• FRT (1)

Note

This enhancer trapping construct is based on pDestTol2CG3, a multisite-Gateway-based plasmid.  It contains a minimal adenovirus E1b promoter driving Gal4VP16 (Gal4 DNA-binding domain fused to the VP16 transcriptional activation domain), followed by the late SV40 polyA signal sequence. Both the Gal4VP16 and SV40 poly A are flanked by FRT sites suitable for Flpe mediated recombination.  Following the 3' FRT site is a Cre ORF and polyA, such that excision of the Gal4VP16 by flpe allows Cre to be expressed under the control of the trapped enhancer.  Following (3’ to) the enhancer trapping construct is a transgene marker: the cmlc2 promoter driving EGFP followed by the SV40 late polyA signal sequence; this marks the heart with green fluorescence.  The enhancer trapping construct and the transgene marker are in a tail-to-tail orientation such that they drive transcription in opposite directions.  The entire cassette (enhancer trapping construct + transgene marker) is flanked by miniTol2 transposon ends (Balciunas et al., 2006).