Part:BBa_K3402054

Over-expression SBLE cassette

This device is composed of upSBLE(BBa_K3402014), Six site(BBa_K3402032), Pgalk(BBa_K3402033), Rec(BBa_K3402034), Tgki(BBa_K3402019), hph(BBa_K3402012), Ptef1(BBa_K3402007), SBLE(BBa_K3402010), Tsyn7(BBa_K3402001), doSBLE(BBa_K3402031).

Usage and Biology

upSBLE and doSBLE are homologous arms, which means this device will edit the SBLE site. The β-Rec/six self-excising system will help to recycle the hygromycin resistance gene. SBLE is the key gene in changing the configuration of sophorolipid from acid type to lactone type.
When we add this fragment to the Cas9 expression device, we will use promoters with different expression strength as we characterized before to control the expression level of UGTB. Then we can produce different amount of lactone-type sophorolipids.
When the over-expression of SBLE and UGTB both work in the cell, there will be two type of sophorolipids produced by our engineering yeast, acid type and lactone type. Different types of sophorolipids have different functions. The combination of lactone type and acid type may increase the effect of sophorolipid. So, we can control the ratio of lactone and acid type sophorolipids.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 858
  Illegal XbaI site found at 2596
  Illegal XbaI site found at 6613
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1380
  Illegal BamHI site found at 3783
  Illegal XhoI site found at 2214
  Illegal XhoI site found at 4935
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 858
  Illegal XbaI site found at 2596
  Illegal XbaI site found at 6613
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 858
  Illegal XbaI site found at 2596
  Illegal XbaI site found at 6613
  Illegal NgoMIV site found at 523
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 5922