Part:BBa_K2350021

R0010-B0034-Holin-B0010-B0012-J23106-B0034-Endolysin-B0010-B0012-J23118-B0034-NrtA-B0015

Endolysin-Holin-NrtA

　　In this part, we combine endolysin (BBa_K2350020), holin (BBa_K2350019), and NrtA (BBa_K2350004). NrtA protein can stick to periplasmic membrane through a flexible linker to capture nitrite or nitrate in the periplasm. Therefore, we make use of NrtA in nitrogen starvation in our 2017 NYMU project. After we reach our goal, we need to kill E.coli to prevent contamination. When the lactose is added into the environment, the suicide mechanism, endolyin and holin, is induced. For more detail information, please see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, BBa_K2350019 for holin part, and BBa_K2350020 for endolysin-holin part.

Promoter, RBS, and Terminator

　　For more details of the connected promoter, RBS, and terminator, you can see in BBa_2350004 for NrtA part, BBa_K2350018 for endolysin part, and BBa_K2350019 for holin part.

Result

Figure 1 is restriction enzyme check electrophoresis result of Endolysin-Holin-NrtA construct. We use BamHI to digest sample 1~3. The result shows that 2 and 3 are right. M represents 1 kb marker.

Figure 1

Sequence and Features

Assembly Compatibility:

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

  Illegal NheI site found at 1035
  Illegal NheI site found at 1058
  Illegal NheI site found at 1757
  Illegal NheI site found at 1780
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BglII site found at 2990
  Illegal BamHI site found at 2360
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 1362
  Illegal AgeI site found at 1432
  
• 1000
  COMPATIBLE WITH RFC[1000]