Part:BBa_K3726069

Lv.1 SigB - B

This is a composite part that corresponds with a level 1 MoClo part in accordance with the marburg collection standard. It is a transcriptional unit for the expression of the part “BBa_K3726063” L0_SigB which codifies a sigma factor that when it is over-expressed increases tolerance to temperature, and reduces ROS stress. Within our project we have used SigB to reduce ROS stress generated from n-butanol.

The part in its upstream region, has the connector 5'Con3 “K2560067” that will allow the assembly of a Lv2 construct following the marburg collection standard. In its downstream region has a homology region “BBa_K3726109” 3CON5(H)_NS3(mod)-down (PCC 11801) for homologous recombination within the genome of PCC 11801.

Expression is controlled by the medium-strong synthetic constitutive promoter J23119 “BBa_K2560031”and the strong synthetic RBS, RBS_PtaBs “BBa_K3726092”

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 850
  Illegal PstI site found at 1752
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 850
  Illegal NheI site found at 431
  Illegal NheI site found at 454
  Illegal NheI site found at 1477
  Illegal PstI site found at 1752
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 850
  Illegal XhoI site found at 886
  Illegal XhoI site found at 1123
  Illegal XhoI site found at 1905
  Illegal XhoI site found at 2025
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 850
  Illegal PstI site found at 1752
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 850
  Illegal PstI site found at 1752
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal SapI.rc site found at 610