Part:BBa_K318000

pBad + Cre + pLac + Hin + pCons + lacI

This composite part is an inducible recombinase generator. It can produce the Cre recombinase if induced with arabinose, and it can produce the Hin recombinase if induced with IPTG or an allo-lactose equivalent. It was refered to as the "Key Cassette" or "Key Construct" by the [http://2010.igem.org/Team:Wisconsin-Madison Wisconsin-Madison 2010] iGEM team.

It can work in conjunction with the Lock Cassette (after junk DNA adjusts the lock to the optimal length). The correct "combination" is IPTG induction and then arabinose induction, expressing the Hin and Cre recombinases respectively.

Note: The attached consititutive lacI expression in this part causes cells transformed with it to grow slowly and in some cases maintain lower intracellular plasmid copy numbers. This is because the promotor, BBa_J23100, is a very powerful constitutive promoter.

A zip file including information about this part is located here: File:BBa K318000 pSB1C3.zip

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1252
  Illegal NheI site found at 3141
  Illegal NheI site found at 3164
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 4342
  Illegal BamHI site found at 1191
  Illegal BamHI site found at 1642
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1026
  Illegal AgeI site found at 1328
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI site found at 1008