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Fig. 3 | Mobile DNA

Fig. 3

From: Compensating for over-production inhibition of the Hsmar1 transposon in Escherichia coli using a series of constitutive promoters

Fig. 3

Papillation assay with a featureless DNA constitutive promoter. a. The Hsmar1 gene is fused to 3x FLAG-tag on its C-terminus and cloned downstream of pEE containing a ribosome binding site (RBS) based on the GACT repeat (RBS+), on an optimal RBS sequence (RBS++), or on an inactive RBS sequence (RBS-). The construct is located between terminator sequences (T) upstream and downstream to avoid read-through transcription. The plasmid backbone is a single-copy vector, pBACe3.6. b. Representative colonies of each single-copy vector expressing a wild-type FLAG-tagged Hsmar1 transposase under the control of pEE with three different RBSs (0 = no transposase/vector only control; pRC1821, 1833 and 1845, negative control: pRC1806). c. Quantification of the number of papillae per colony from single colonies. Average ± standard deviation of six representative colonies from the same biological replicate

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