Team:BostonU/Parts

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OUR PARTS

We submitted two composite parts to the registry: BBa_K2411005 and BBa_K2411008.

 

BBa_K2411005: Forward Engineered Toehold 1 with deGFP gene

This device encodes for a toehold that when activated expresses deGFP. It has an OR2-OR1 promoter, followed by the first forward engineered toehold sequence designed by Green et al. The deGFP follows the toehold, and the device ends with a T7 terminator. The part has been designed for and tested in an E. coli derived transcriptional translation cell free system. In order for the part to function, it needs to be used in conjunction with a trigger RNA. Characterization data can be view on our results page or on the registry.

 

BBa_K2411008: Forward Engineered Toehold 2 with deGFP gene

This device encodes for a toehold that when activated expresses deGFP. It has an OR2-OR1 promoter, followed by the second best forward engineered toehold sequence designed by Green et al. The deGFP follows the toehold, and the device ends with a T7 terminator. The part has been designed for and tested in an E. coli derived transcriptional translation cell free system. In order for the part to function, it needs to be used in conjunction with a trigger RNA. Characterization data can be view on our results page or on the registry. 

In addition to these composite parts, we created registry pages for the OR2-OR1 promoter, the T500 terminator, and both toehold architectures that we used in our composite parts.

 

BBa_K2411000: OR2-OR1 Promoter

This is a constitutively active promoter that can be used in E. coli and E. coli derived transcription translation systems. This part is derived from the lambda bacteriaphage.

 

BBa_K2411002: T500 Terminator

This is a bacterial terminator.

 

BBa_K2411004: Toehold 1

This sequence is the forward engineered toehold from Green et. al. This part contains the hairpin loop sequence that with the RBS. There is no gene associated with this toehold.

 

BBa_K2411007: Toehold 2

This sequence is the second best performing forward engineered toehold from Green et. al. There is no gene associated with this toehold.

 

In addition, the deGFP part utilized in our composite part was submitted under BBa_K2205001 by iGEM17_Newcastle.

 

The sequences for the promoter, terminator, and deGFP were obtained from the pBEST plasmid used to Vincent Noireaux and his lab with their cell-free system [1].

 

The sequences for the toeholds were obtained from Green et al. [2].

A toehold switch part including a ribosomal binding site and a start codon.

 

[1]Shin, Jonghyeon, and Vincent Noireaux. "Efficient cell-free expression with the endogenous E. Coli RNA polymerase and sigma factor 70." Journal of biological engineering 4.1 (2010): 8.

 

[2] Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939.