Part:BBa_K4805011

pTEF2-ERG20_F96W-tSSA1-pPGK1-ClSS_ S532A,F441V-tPGK1

description

ERG20 catalyzes the sequential formation of geranyl diphosphate (GPP) and farnesyl diphosphate (FPP). ERG20_F96W, as a F96W mutant can help increase the supply of FPP. ClSS_S532A,F441V encodes for a variant of ClSS, a santalene synthase from Clausena lansium(Genbank Accession: ADR71055.1), which possesses two site direct mutagenesis of S532A and F441V. Our part can be a reference for other teams to perfect the pathway producing santalene from FPP. It belongs to the terpenoid part collection we have established for the production of santalol and ambrein in S. cerevisiae, which includes BBa_K4805000-BBa_K4805012.

usage and biology

==sequencing and features

Assembly Compatibility:

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

  Illegal BglII site found at 1302
  Illegal BglII site found at 1363
  Illegal BglII site found at 2828
  Illegal BglII site found at 3917
  Illegal XhoI site found at 1479
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1018
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1465
  Illegal BsaI site found at 1726
  Illegal BsaI.rc site found at 1715
  Illegal BsaI.rc site found at 2838
  

characterization

2023 LINKS-China chose a commonly used 106 site to insert ERG20_F96W and ClSS at the same time(Figure 1A). It is obvious that we successfully screened one positive strain out of 10 strains, which is named as Lv2s-1 (Figure 1B). Same as ClSS, ClSS_F441V, S532A was also inserted into 106 site.