Part:BBa_K2213004

PduD(1-20)_cgPPK2_mCherry_His6

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Usage and Biology

Fusion of the PduD(1-20), Corynebacterium glutamicum class II polyphosphate kinase and mCherry followed by a hexahistidine tag.

The construct was heterologously expressed in a BL21 (DE3) strain of E. coli under the control of a T7 promoter. The construct was cleaved before or during purification, resulting in the elution of a ~30 kDa (expected wieght: 66 kDa), his-tagged protein with a bright pink colour from the Ni-NTA agarose media (Figure 1). The protein fragment was not tested for kinase activity.

</h3>Figure 1. SDS-PAGE analysis of fractions from the Ni-NTA agarose purification of the PduD(1-20)_cgPPK_mCherry construct. The black arrow indicates where the expected band should be (66 kDa) in the elution fraction lane. Ladder: Precision Plus Protein™ (Biorad)

</br> Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 148
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 1030
  Illegal SapI.rc site found at 827