We added a total of six new basic parts to the iGEM registry (BBa_K4660000 - BBa_K4660005).
| Number | Type | Description | Length |
|---|---|---|---|
| BBa_K4660000 | Cloning | “vbrK” This is the coding sequence for the histidine kinase VbrK from Vibrio parahaemolyticus. This part was codon optimized for gene expression in E. coli. | 1452bp |
| BBa_K4660001 | Cloning | “vbrR” This is the coding sequence for the response regulator VbrR from Vibrio parahaemolyticus. This part was codon optimized for gene expression in E. coli. | 660bp |
| BBa_K4660002 | Cloning | “blaR1” This is the coding sequence for the sensor metalloprotease BlaR1 from Staphylococcus aureus. This part was codon optimized for gene expression in E. coli. | 1755bp |
| BBa_K4660003 | Cloning | “blaI” This is the coding sequence for the transcription repressor BlaI from Staphylococcus aureus. This part was codon optimized for gene expression in E. coli. | 387bp |
| BBa_K4660004 | RBS | “promotor-VbrR” This is our predicted promotor region from Vibrio parahaemolyticus to which the activated response regulator VbrK binds and activates gene transcription. | 302bp |
| BBa_K4660005 | RBS | “promotor-BlaI” This is the endogenous promotor sequence from Staphylococcus aureus where BlaI dimers bind and repress gene transcription. | 83bp |
To use these basic parts for the design of our biosensor we ordered the genes from Twist Bioscience and added compatible BsaI restriction sites, so we could use them directly for a Golden Gate assembly into a level 1 vector. We ordered them in the standard high-copy vector from Twist Bioscience, here called pTWIST.
Figure 1: Basic parts for Golden Gate assembly with VbrK/VbrR and the respective promotor sequence.
Figure 2: Basic parts for Golden Gate Assembly with BlaR1/BlaI and the respective promotor sequence.