Difference between revisions of "Part:BBa K4268016:Design"
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The Neck and Tail Multi-Transcriptional Unit (MTU) codes for the putative proteins that makeup the neck region (Tail tubular A, B, and the Head-tail connector protein) and tail fibers (Tail-fiber like protein) of S-Tip37. The component transcriptional units and basic parts were designed to be compatible with Type IIS assembly via Golden gate cloning. The MUT is a level 2 assembly of multiple level 1 transcriptional units' and permits modular assembly so the user may customize the part as needed. The component transcriptional unit parts have been uploaded to the parts repository under the parts numbers BBa_K4268008, BBa_K4268012, BBa_K4268013, and BBa_K4268015. | The Neck and Tail Multi-Transcriptional Unit (MTU) codes for the putative proteins that makeup the neck region (Tail tubular A, B, and the Head-tail connector protein) and tail fibers (Tail-fiber like protein) of S-Tip37. The component transcriptional units and basic parts were designed to be compatible with Type IIS assembly via Golden gate cloning. The MUT is a level 2 assembly of multiple level 1 transcriptional units' and permits modular assembly so the user may customize the part as needed. The component transcriptional unit parts have been uploaded to the parts repository under the parts numbers BBa_K4268008, BBa_K4268012, BBa_K4268013, and BBa_K4268015. | ||
| − | [[File: T--SUNY Oneonta--neck-tail MTU.png| | + | [[File: T--SUNY Oneonta--neck-tail MTU.png|500px|thumb|center|Figure 1: SBOL representation of the Neck and Tail MTU]] |
One of the main concerns in designing the level 2 MTUs is gene order. Studies have demonstrated that the order that genes are placed within an operon can impact the rate of translation, and therefore, the level of expression (Wells et. al. 2016, Lim et. al. 2011). While each gene in the MTU will have its own promoter, and is therefore not organized within an operon, gene order may impact expression, and will likely require optimization. For the initial iteration of the MTU design we ordered the genes in the MTU so that genes requiring the highest levels of expression were not located on the “ends” of the MTU. | One of the main concerns in designing the level 2 MTUs is gene order. Studies have demonstrated that the order that genes are placed within an operon can impact the rate of translation, and therefore, the level of expression (Wells et. al. 2016, Lim et. al. 2011). While each gene in the MTU will have its own promoter, and is therefore not organized within an operon, gene order may impact expression, and will likely require optimization. For the initial iteration of the MTU design we ordered the genes in the MTU so that genes requiring the highest levels of expression were not located on the “ends” of the MTU. | ||
Revision as of 16:22, 7 October 2022
Neck and Tail Multi-Transcriptional Unit
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 1760
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 398
Illegal NheI site found at 4934
Illegal PstI site found at 1760 - 21COMPATIBLE WITH RFC[21]
- 23INCOMPATIBLE WITH RFC[23]Illegal PstI site found at 1760
- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 1760
Illegal NgoMIV site found at 6431
Illegal AgeI site found at 8462 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The Neck and Tail Multi-Transcriptional Unit (MTU) codes for the putative proteins that makeup the neck region (Tail tubular A, B, and the Head-tail connector protein) and tail fibers (Tail-fiber like protein) of S-Tip37. The component transcriptional units and basic parts were designed to be compatible with Type IIS assembly via Golden gate cloning. The MUT is a level 2 assembly of multiple level 1 transcriptional units' and permits modular assembly so the user may customize the part as needed. The component transcriptional unit parts have been uploaded to the parts repository under the parts numbers BBa_K4268008, BBa_K4268012, BBa_K4268013, and BBa_K4268015.
One of the main concerns in designing the level 2 MTUs is gene order. Studies have demonstrated that the order that genes are placed within an operon can impact the rate of translation, and therefore, the level of expression (Wells et. al. 2016, Lim et. al. 2011). While each gene in the MTU will have its own promoter, and is therefore not organized within an operon, gene order may impact expression, and will likely require optimization. For the initial iteration of the MTU design we ordered the genes in the MTU so that genes requiring the highest levels of expression were not located on the “ends” of the MTU.
For assembly, the order in which TUs will be assembled into MTUs will be dependent on the 5' and 3' flanking sites present on each TU. These sites are provided by the level 1 vector. Therefore, for our level 2 MTU design it will be critical to clone the level 1 parts into the appropriate level 1 vector. To obtain this design, the level one parts should be assembled into the iGEM standard golden gate vectors as in table 1.
For the same reasons, when assembling the level 2 component, care should be taken to select vectors that will yield the desired component order.
Source
The coding sequences were derived from the S-TIP37 genome.