Team:MIT/Basic Part

Basic Parts

Team MIT has chosen our cpComE-NLS-VP16-NES SM6 construct to share as our best part. This part contains a human optimized coding sequence for a constitutively phosphorylated ComE, the putative response regulator protein for the ComCDE two-component signaling pathway, native to Streptococcus mutans strain SM6 (a bacteria common in the human oral cavity). Design elements were chosen to optimize expression and function of ComE in human cells. The aspartic acid at amino acid position 60 (the putative phosphorylation site of the response regulator) in a wildtype ComE was changed to a glutamic acid, to act as a phosphomimetic.

Normally, to be used as part of a functional sensing system, the ComE response regulator must be paired with the ComD histidine kinase receptor protein. When ComD binds to the S. mutans competence stimulating peptide, it autophosphorylates and transfers a phosphate group to an aspartic acid on ComE, enabling ComE to oligomerize and bind to specific DNA sequences to promote transcription. This constitutively phosphorylated ComE should theoretically always be able to bind DNA. The cpComE coding sequence is fused to a nuclear localization sequence (NLS), VP16 activating domain, and a nuclear export sequence (NES). It is in a pSB1C3 backbone and is BioBrick and MoClo compatible.

Please find below the list of all of our basic parts!

Name	Type	Description
BBa_K2649001	Coding	cpComE-NLS-VP16-NES SM6
BBa_K2649000	Coding	ComE-NLS-VP16-NES SM6
BBa_K2649004	Coding	ComE-NLS-VP16-NES NN2025
BBa_K2649005	Coding	cpComE-NLS-VP16-NES NN2025
BBa_K2649010	Coding	cpComE-NLS-VP16-NES UA159
BBa_K2649011	coding	eComE-NLS-VP16-NES
BBa_K2649006	Regulatory	ComE Binding Promoter Low Affinity
BBa_K2649007	Regulatory	ComE Binding Promoter Med Affinity
BBa_K2649008	Coding	CD4-ComD-mKO2 NN2025
BBa_K2649009	Coding	CD4-ComD-mKO2 UA159
BBa_K2649002	Backbone	Phytobricks Universal Backbone with RFP TU Insert
BBa_K2649003	Backbone	pSB1K3 with RFP TU Insert