Part:BBa_K1979004:Design

GFP-PBP5

Assembly Compatibility:

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

  Illegal NheI site found at 7
  Illegal NheI site found at 135
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1892
  Illegal BamHI site found at 168
  Illegal XhoI site found at 2100
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1206
  Illegal AgeI site found at 1808
  
• 1000
  COMPATIBLE WITH RFC[1000]

Design Notes

Histidine tag is added for affinity purification to harvest the proteins. g10-L is a strong RBS in E. coli but also well suited for foreign gene expression.The effect of the ribosome binding site (RBS) on in vitro translation has been investigated extensively (Karig et al. 2012; Lentini et al. 2013; Takahashi et al. 2013). Most important for an efficient reaction is a defined distance (4-9 bases) of the RBS to the following start codon (Karig et al. 2012; Lentini et al. 2013), but the strength of the RBS has a minor effect as long as it is not too weak (Karig et al. 2012; Takahashi et al. 2013).

Source

The GFP sequence is cloned from eGFP plasmid, PBP5 sequence is from BBa_K1979003, and the promoter comes from BBa_J23101.