Week 2 HW: Genetic Circuit Design for Deinoxanthin

Week 2 Homework – Genetic Circuit Design

Part 0: Kernel Setup

Repository: awais-deinoxanthin
Folder: awais-deinoxanthin

Part 1: Expression Target

I designed a genetic construct to increase deinoxanthin production in Deinococcus radiodurans.

  • Molecules: Two enzymes – CrtB (phytoene synthase) and Dxs
  • Host: Deinococcus radiodurans
  • Location: Cytoplasm

Part 2 & 3: Genetic Parts and Construct

I created the following parts in Kernel:

  • Promoter: P_constitutive_Deinococcus
  • RBS: RBS_strong
  • CDS: crtB_Deinococcus
  • RBS: RBS_strong_2
  • CDS: dxs_Deinococcus
  • Terminator: Terminator_strong

Construct name: Deinoxanthin_Expression_Cassette

Order of parts:
Promoter → RBS → crtB → RBS → dxs → Terminator

Part 4: Optimize and Quality Check

I codon-optimized the coding sequences for Deinococcus.
I checked the sequences using BLAST and ran the safety screen. No high-risk sequences were found.

Part 5: From Design to Build

For the first version, I will order the complete construct as synthetic DNA.
If multiple versions are needed later, I can use Gibson assembly. Deinoxanthin Expression Cassette Deinoxanthin Expression Cassette

Part 6: What I don’t know yet

  1. Will the promoter work well in Deinococcus radiodurans?
  2. Will the cells grow normally after overexpressing these two genes?
  3. How much extra deinoxanthin will be produced?

AI Note
I used Grok to help structure the design and understand the Kernel workflow. The final decisions and understanding are my own.