Conclusions of the experimental data

We have constructed the pINDEL plasmid.  We have observed that this construct is stable at 30°C and unstable at 42°C, as expected.  The DEL activity of pINDEL was shown to be functional as we were able to excise a FRT’-Cm-FRT’ cassette inserted in the E. coli chromosome at very high frequency.  Unfortunately, our assay for the IN activity failed.  More tests will be needed to optimize the IN assay.

In addition to the pINDEL construction, we rebuild a functional biobrick containing the FRT’-CmR-FRT’ cassette (BBa_K551000).  The biobrick is functional since it confers Cm resistance.  Moreover, we were able to show that the FRT’ sites of the intermediate construct (TOPO®-FRT’-Cm-FRT’) are functional since the Cm resistance phenotype is lost upon FLP induction, although at low frequency.