Part:BBa_K1861401:Design
GS-Linker and rpd3
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 222
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This is a component of BBa_K1861040.
In order to clone rpd3 into BBa_K1861010, we had to add the restriction site for SacI to the rpd3 gene. We used the following primer to achieve this:
3' SacI primer - ACA TGC TCA GAG CTC TCC ACT GC
The restriction site is in italic.
To synthesize the rpd3 gene, we ordered both halves of it from IDT (the first half also included BBa_K1159200) and fused them together by an overlap-PCR. The following sequence (102 nucleotides) was used as overlap:
GCTGTCAACTATGCGGGTGGTTTGCATCATGCAAAAAAATCGGAAGCATCTGGGTTTTGTTATTTAAATGACATAGTACTGGGCATTATTGAGCTACTACGA
Source
The rpd3 gene is located on chromosome XIV of S. cerevisiae. Its sequence was obtained through the Saccharomyces Genome Database.
References
https://eu.idtdna.com/site, visited on 2015-09-19
http://www.yeastgenome.org/locus/S000005274/overview, visited on 2015-09-21
http://web.archive.org/web/20150921180338/http://www.yeastgenome.org/locus/S000005274/overview