![](https://parts.igem.org/images/partbypart/icon_coding.png)
Part:BBa_K2271107:Design
PEX5 variant R19 with Promotor/Terminator
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1341
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1128
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1633
- 1000COMPATIBLE WITH RFC[1000]
Part design
This part was designed in the course of a random mutagenesis approach. With the predicted three dimensional strucutre of the TPR domains we carried out our workflow in order to obtain the PEX5 variant we wanted.
After gene synthesis we performed golden gate cloning and made it compatible to the biobrick standard via overhang PCR with the appropriate primers. This product then was ligated into the biobrick backone pSB1C3.
Source
The original sequence was taken from NCBI (GenBank: KZV12481.1). After mutagenesis we performed the IDT codon optimization for Saccharomyces cerevisiae, removed any restrictions sites that would interfere with the biobrick or golden gate standard and synthesized the gene with appropiate Golden Gate overhangs.