Part:BBa_K3407018:Design

Mini-3 endoribonuclease with araBAD promoter - RBS - and rrnBT1 terminator

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 300
  Illegal XbaI site found at 673
  Illegal PstI site found at 527
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 300
  Illegal PstI site found at 527
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 300
  Illegal BglII site found at 309
  Illegal BamHI site found at 809
  Illegal XhoI site found at 818
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 300
  Illegal XbaI site found at 673
  Illegal PstI site found at 527
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 300
  Illegal XbaI site found at 673
  Illegal PstI site found at 527
  Illegal AgeI site found at 74
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 56
  

Design notes

The biobrick is made with EcoRI and BglII before the RBS and CDS, and BamHI and XhoI after the CDS, so the piece CDS can be excised using these set of enzymes.

Source

Genomic DNA from B. subtilis strain 168. Plasmid used pBbE8c backbone from BglBricks. pBbE8c-RFP was a gift from Jay Keasling (Addgene plasmid # 35269; http://n2t.net/addgene:35269; RRID:Addgene_35269) [1] Please refer to the Addgene page for more information about licences associated with the use of the plasmid.

References

1. BglBrick vectors and datasheets: A synthetic biology platform for gene expression. Lee TS, Krupa RA, Zhang F, Hajimorad M, Holtz WJ, Prasad N, Lee SK, Keasling JD. J Biol Eng. 2011 Sep 20;5:12. 10.1186/1754-1611-5-12 PubMed 21933410