Difference between revisions of "Part:BBa K2271105:Design"
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===Part design=== | ===Part design=== | ||
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| − | This part was designed in the course of a | + | This part was designed in the course of a targeted 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. |
<br> | <br> | ||
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 <b>pSB1C3</b>. | 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 <b>pSB1C3</b>. | ||
Latest revision as of 18:53, 1 November 2017
PEX5 variant R15
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 409
- 1000COMPATIBLE WITH RFC[1000]
Part design
This part was designed in the course of a targeted 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.