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t7-header-icon Voigt Lab Fragmented T7 RNAP Plasmids


Christopher Voigt's lab has built a ‘resource allocator’ to control synthetic genetic systems by splitting T7 RNA polymerase (RNAP) variants into a number of fragments. By tuning the production of a limiting protein fragment, the total number of polymerases in a synthetic system can be controlled while allowing the allocation of this resource between different outputs.

A publication from the Voigt lab in Molecular Systems Biology (Segall-Shapiro TH, et al., MSB, 2014) describes the bisection mapping of T7 RNAP to produce functional protein fragments and the composition of these fragments into a resource allocator system. Four orthogonal T7 RNAP variants were bisected into a conserved ‘core fragment’ and variable ‘σ fragments’. T7 RNAP is further shown to tolerate fragmentation into three and four fragments while retaining function. The three-piece RNAP and further mutations are used to create accessory fragments for further regulation of the resource allocator.

Voigt Lab Fragmented T7 RNAP Plasmids

The work and corresponding plasmids are described in:

A ‘resource allocator’ for transcription based on a highly fragmented T7 RNA polymerase. . Segall‐Shapiro TH, Meyer AJ, Ellington AD, Sontag ED & Voigt CA. Molecular Systems Biology. 2014 July 30. doi: 10.15252/msb.20145299. PubMed.

The Voigt lab has deposited plasmids encoding the fragmented T7 RNAP system that can be used to implement resource allocation. These include orthogonal T7 RNAP variant reporter plasmids (pTHSSd_8 – 11), plasmids with inducible σ fragments (pTHSSd_23 – 26), core fragment expression plasmids (pTHSSd_38 – 40), ‘accessory’ fragments (pTHSSd_27, 29, 42), and control plasmids (pTHSSd_34, 41, 44). They have also deposited plasmids to express three- and four-piece T7 RNAP (pTHSSd_14, 18) and a modified transposon that can be used to map the tolerated bisection sites in a protein (pTHSSd_1).

Resources

Individual plasmids can be ordered via the links below:

ID Plasmid
59947 pTHSSd_1 Add to Cart
59948 pTHSSd_8 Add to Cart
59949 pTHSSd_9 Add to Cart
59951 pTHSSd_11 Add to Cart
59952 pTHSSd_14 Add to Cart
59953 pTHSSd_18 Add to Cart
59954 pTHSSd_23 Add to Cart
59955 pTHSSd_24 Add to Cart
59956 pTHSSd_25 Add to Cart
59957 pTHSSd_26 Add to Cart
59958 pTHSSd_27 Add to Cart
59959 pTHSSd_29 Add to Cart
59960 pTHSSd_34 Add to Cart
59963 pTHSSd_40 Add to Cart
59966 pTHSSd_44 Add to Cart