Colicin E2 Immunity protein under pLac promoter (BBa_R0011) with repressilator RBS. This part can be used to confer immunity against the colicin E2 activity protein by induction with IPTG.
We selected the repressilator RBS and pLac to drive production of Colicin E2 Immunity protein
Rnase Ba (Barnase) from Bacillus amyloliquefaciens
Rnase Ba (Barnase) from Bacillus Amyloliquefaciens
ptrc* is a modified form of ptrc promoter with the -35 and -10 regions being present in the reverse order and the promoter is flanked by fimE recognition sequence. FimE is an invertase capable of inverting any DNA sequence flanked between its recognition seqence. This fim inversion system would help avoid leaky expression. We expressed a new lysis device, DpnI under the control of fim inversion promoter. DpnI is a restriction enzyme capable of chopping up all methylated DNAs. We propose DpnI as a novel lysis device. DpnC and DpnD are obtained from Streptococcus pneumoniae R6 strain.
One should be careful while expressing Dpn genes. Dpn is supposed to chop up the DNA and cause cell death. Hence, no leaky level of Dpn is needed to recover live cells
repressilator RBS Colicin E2 Immunity protein
Colicin E2 Immunity protein with the repressilator RBS
This part contains the ColE2 operon in its entirety.
This part was cloned from a plasmid containing the operon which was received from UC Berkeley.
ColicinE1 Producer Controlled by 3OC6HSL Receiver Device
Pascal Kraemer, Marika Ziesack, Kathrin Nussbaum and Andreas Kuehne
The constitutive expressed LuxR protein is able to build a complex with the AHL molecule. This complex function as transcription factor for the lux pR promoter and activates by this the colicinE1 production. This activation of the lux pR promoter can additionally lead to expression of the kil protein by read through of the terminator t1 and therefore to lysis of the cell and colcin release. The colicinE1 immunity protein (imm) lying on the reverse strand is expressend under a constitutive promotor.