Part:BBa_K243033

pEX

This is our main E. coli protein expression vector. To insert DNA fragments, the plasmid possesses the restriction sites XbaI NgoMIV and AgeI_SpeI_NotI_PstI. The IPTG-inducable plasmid vector contains the pBR322 origin of replication and a Shine-Dalgarno-Sequence.

Usage and Biology

We used this vector containing an ampicillin resistance gene for the expression of single constructs such as the His-FluA-Split-Fok_i (BBa_K243010)construct. Simultaneous expression of the other Fok construct, His-Dig-Split-Fok_a (BBa_K243036) could be assured using the vector pJS419 since it confers chloramphenicol resistance to the host cell. This enables a positive selection of bacterial strains such as XL1-blue (Stratagene) cotransformed with both plasmids.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal suffix found in sequence at 2734
  Illegal XbaI site found at 1523
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal SpeI site found at 2735
  Illegal PstI site found at 2749
  Illegal NotI site found at 2742
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 1808
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal suffix found in sequence at 2735
  Illegal XbaI site found at 1523
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal suffix found in sequence at 2725
  Illegal XbaI site found at 1523
  Illegal NgoMIV site found at 1531
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 3800
  Illegal SapI.rc site found at 4799