Part:BBa_K3739051

J23100-B0030-LamB-LC1KR-2-GFP-B0010

We anchor LCIKR-2 protein onto membranes through LamB to stick the engineered bacteria on the polypropylene. We use BBa_K3739051 to construct the expression system and anchor LCIKR-2 on the surface of VnDX.

Biology

LamB is an anchor protein from Vibrio Species, which can anchor its passenger protein to the cell membrane. It has been widely used in cell-surface display. LCIKR-2, A mutant of a peptide called LCI from Bacillus subtilis, can sticks to polypropylene strongly. LCIKR-2 is fused at C terminal with LamB so that LCIKR-2 can be displayed on the surface of VnDX.

Characterization

1. Identification

After receiving the synthesized DNA, PCR was done to certify that the plasmid was correct, and the experimental results were shown in figure1. ""

Fig. 1. Gene circuit and agarose gel electrophoresis. (A) Gene circuit of J23100-B0030-LamB-LC1KR-2-GFP-B0010 (BBa_K3739051). (B)Target bands of LamB-LC1KR-2-GFP (black arrow, 2300 bp).

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 7
  Illegal NheI site found at 30
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 592
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 740
  Illegal SapI.rc site found at 1496