Part:BBa_K3726071

Lv.1 AcrbV2 - 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_K3726065” L0_AcrbV2 which codifies a membrane transporter subunit , which has been optimized for the specific secretion of n-butanol. Within our project we have used AcrbV2 to enhance n-butanol secretion through cell membrane in order to cut down on toxicity generated by itself.

The part is flanked by two connectors that will allow the assembly of a Lv2 construct. In its upstream region the connector is 5'Con2 Bba_K2560066, and in its downstream region 3'Con2 Bba_K2560071 .

Expression is controlled by the promoter PgntK “BBa_K3726078” which is a negative feedback membrane stress promoter that has demonstrated that can optimize the expression levels of AcrB drug efflux pump to maximize the growth rate and minimize cellular toxicity.

Moreover, the expression is controlled as well by the strong synthetic RBS, RBS_PtaBs “BBa_K3726092”.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 1023
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 752
  Illegal PstI site found at 1023
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 1023
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 1023
  Illegal NgoMIV site found at 2159
  Illegal NgoMIV site found at 3675
  Illegal AgeI site found at 109
  
• 1000
  COMPATIBLE WITH RFC[1000]