Part:BBa_K1074001

PHT43

Usage and Biology

PHT43 is a E.coli-B.subtilis shuttle vector allowing high-level expression of secreted proteins in B.subtilis. It is based on the strong σA-dependent promoter preceding the groE operon of B. subtilis which has been converted into an efficiently controllable (IPTGinducible)promoter by addition of the lac operator. An efficient Shine-Dalgarno (SD) sequence as well as a multiple cloning site (BamH I, Xba I, Aat II, Sma I) were also inserted. To obtain secretion of recombinant proteins, the coding region for the signal peptide of the amyQ gene encoding an α-amylase was fused to the SD sequence.

Vector Map of pHT43

Reference

<Bacillus subtilis Expression Vectors> MoBiTec GmbH 2012

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 1255
  Illegal EcoRI site found at 3507
  Illegal XbaI site found at 2911
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 1255
  Illegal EcoRI site found at 3507
  Illegal NheI site found at 1091
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 1255
  Illegal EcoRI site found at 3507
  Illegal BglII site found at 2978
  Illegal BglII site found at 6059
  Illegal BamHI site found at 2905
  Illegal XhoI site found at 6063
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 1255
  Illegal EcoRI site found at 3507
  Illegal XbaI site found at 2911
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 1255
  Illegal EcoRI site found at 3507
  Illegal XbaI site found at 2911
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 6954
  Illegal SapI.rc site found at 4131
  Illegal SapI.rc site found at 8036