Team:ETH Zurich/Parts

Parts

Overview

We submitted in total 14 new BioBricks to the iGEM registry! You can find all parts and their design below.

Basic Parts

Part Description BioBrick
Bacterioferritin Heme-deletion mutant of bacterial iron storage protein functioning as MRI contrast agent BBa_K2500000
Azurin Mature cytotoxic copper protein BBa_K2500001
p28 Effector domain of Azurin BBa_K2500002
PtlpA Best Basic Part: Temperature-responsive promoter TlpA optimized for slight activation above 37°C and full activation at 45 °C BBa_K2500003
TlpA Protein reversibly inhibiting the pTlpA promoter at temperatures below 37 °C BBa_K2500004
AND gate a Synthetic promoter responsive to LldR and luxR BBa_K2500010
AND gate b Synthetic promoter responsive to LldR and luxR BBa_K2500011
AND gate c Synthetic promoter responsive to LldR and luxR BBa_K2500012

Composite Parts

Part Description BioBrick
Pconst_RBSeng_TlpA Best Composite Part:Temperature-dependent transcriptional repressor TlpA regulated by constant promoter and engineered RBS BBa_K2500008
pTlpA_Protein E Bacterial cell lysing protein E regulated by heat-inducible promoter pTlpA and synthetic RBS BBa_K2500009
Pconst_RBS_LldP/
LldR_Pconst_RBS_LuxR
Expression cassette consisting of LuxR and LldP/LldR BBa_K2500013

Design

BBa_K2500010: AND gate a

In absence of high concentrations of L-lactate, LldR inhibitor proteins bind to the binding sites O1 and O2 surrounding the pLux promoter leading to the formation of a DNA loop. The pLux promoter is sequestered and inaccessible for transcription. In design a, the distances between the intercalated promoter and the binding sites were adapted from BBa_K1847007.

Registry: BBa_K2500010

BBa_K2500011: AND gate b

In design b, each binding site was duplicated in order to achieve a potential zipper mechanism and stronger inhibition due to binding more LldR inhibitor proteins.

Registry: BBa_K2500011

BBa_K2500012: AND gate c

In design c, an artificial spacer was embedded between the pLux promoter and the O2 binding site in order to influence the looping dynamics.

Registry: BBa_K2500012