Part:BBa_K2758000:Design
Mercury activated Killer Red Protein suicide switch
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 237
Illegal BsaI.rc site found at 528
Design Notes
This part is designed to work with another part that consists of a Hg accumulator. This part has been designed to express the Killer Red protein later than the accumulator is expressed from the other construct. In order to delay the expression of Killer Red protein relative to the accumulator, we inserted an RBS with moderate binding capabilities into the K2758000 construct. We built this construct through IDT adding the NotI restriction sites.
Source
This sequence was constructed entirely with Biobricks from the iGEM registry. The sequences are: BBa_K346002 (mercury responsive promotor), BBa_B0030 (RBS), BBa_K1184000 (Killer Red), BBa_B0015 (double terminator). The part was designed using Snapgene and then sent to be synthesized by IDT. The biobrick was made by copying the sequences of the components from their respective registry pages in the order: BBa_K346002, BBa_B0030, BBa_K1184000, and BBa_B0015. An XbaI/SpeI ligation scar was added in between each part to simulate a ligation, however, we replaced the ligation scars flanking the RBS with HindIII and ApaI restriction sites. These restriction sites serve two purposes:
1) To provide a method of checking whether or not the ligation of the biobrick to the PSB1C3 vector is successful
2) To facilitate future characterization of the part with different RBS in order to delay or promote the expression of KR.
Figure 1. Map of the Killer Red construct displayed in the Snapgene software