Part:BBa_K5310061:Design
Transmembrane-Intracellular basic structure for chimeric autoantibody receptor
- 10INCOMPATIBLE WITH RFC[10]Illegal PstI site found at 146
- 12INCOMPATIBLE WITH RFC[12]Illegal PstI site found at 146
- 21COMPATIBLE WITH RFC[21]
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- 25INCOMPATIBLE WITH RFC[25]Illegal PstI site found at 146
Illegal AgeI site found at 380 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
In order to design the transmembrane and intracellular modular part of our chimeric autoantibody receptor, we began by identifying all the sequences of interest (see below) and gathering relevant sequences from databases such as NCBI, UniProt, and the iGEM Registry, while utilizing PDB to analyze the structure of individual domains. We then visualized and refined the structural models using PyMOL and conducted codon optimization via Twist Bioscience to enhance expression. Sequence design and visualization were performed on Benchling, then we used Robetta-Baker Lab and its deep learning-based modeling method, RoseTTAFold for structural modeling. The design process was further informed by recent literature, including studies by Castella et al. (2019), Rafiq et al. (2020), Ellebrecht et al. (2020), and Chang & Chen (2017), along with insights from two previous iGEM teams that developed CAR constructs (iGEM Patras med 2023, iGEM Munich 2022).
Source
The sequences of all individual domains are taken from the human genome (from NCBI) and codon optimised for Homo sapiens.