Part:BBa_K2368017

Introduction

500 bp of homologous arm＋His

General

Sst2 protein is an important negative regulatory factor of the pheromone GPSTP. In order to improve the detection sensitivity, we design this part to knock out the sst2 gene.

Our experiment

Similarly, we design 3 pairs of primers and the marker is Histone synthesis gene, short termed His. We get 3 fragments by PCR and they have the overlap areas with each other. Then, the complete fragment observed by OE-PCR is converted to the yeast. Additionally, the colony is chosen on the relevant nutritional deficiency medium, so that only the positive cloning can survival on it.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1240
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1131
  Illegal BglII site found at 1191
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1801
  
• 1000
  COMPATIBLE WITH RFC[1000]