Part:BBa_K1861401:Design

GS-Linker and rpd3

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal NgoMIV site found at 222
  
• 1000
  COMPATIBLE 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