Part:BBa_K200010

TaqI Composite

This sequence codes for the restriction enzyme TaqI and also includes and RBS and double terminator which will result in a closed transcriptional unit once this construct is ligated downstream of a promoter.

Detail

This is a Type II restriction enzyme that recognises the sequence TCGA. Its activity can be blocked by dam methylation.

The working temperature is 65°C.

As part of the Imperial 2009 iGEM E.ncapsulator project, TaqI was used along with DpnII to cut up the genetic material within the cell, completing the cell death module.

Usage and Biology

The restriction enzyme TaqI, is isolated from the bacterium Thermus aquaticus. 1

It cuts at a site that is near to where early and late messenger RNA transcription starts in opposite directions. Furthermore, it is a 4 base cutter, leaving a 2 base overhang.

The cutting site is as shown:

1

===Sequence and Features===

Assembly Compatibility:

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

  Illegal BglII site found at 36
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 175
  

References

<biblio>

1. 1 pmid=204628

</biblio>

2. [http://www.neb.com/nebecomm/products/productM0219.asp Information from NEB]