Part:BBa_K731710

Platform for terminators analysis under the control of tac promoter

This is a platform for the characterization of terminator efficiency. It was built starting from K731700, where the T7 promoter was replaced with a tac promoter by insertion/deletion. The platform has been successfully used for the ratiometric analysis of terminator efficiency.

The combined use of BBa_K731710 and BBa_K731700 allow also to analyse any potential difference in terminators' activity due to different polymerases

Usage and Biology

This part has been created by the iGEM Trento 2012 team for terminator characterization.

Protocol

The two proteins have partially overlapping emission spectra and different folding kinetics. Suggested wavelengths are:

	Excitation (nm)	Emission(nm)
mCherry	587	610
Venus	515	528

Sequence and Features

Assembly Compatibility:

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

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 6865
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NotI site found at 9
  Illegal NotI site found at 6871
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal EcoRI site found at 6865
  Illegal BglII site found at 6011
  Illegal BamHI site found at 6847
  Illegal XhoI site found at 753
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal prefix found at 6865
  Illegal suffix found at 2
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal prefix found at 6865
  Plasmid lacks a suffix.
  Illegal XbaI site found at 6880
  Illegal SpeI site found at 2
  Illegal PstI site found at 16
  Illegal NgoMIV site found at 714
  Illegal NgoMIV site found at 1051
  Illegal NgoMIV site found at 4231
  Illegal NgoMIV site found at 4391
  Illegal NgoMIV site found at 5979
  Illegal AgeI site found at 6815
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Plasmid lacks a prefix.
  Plasmid lacks a suffix.
  Illegal BsaI site found at 2228
  Illegal SapI.rc site found at 3310