Part:BBa_K5302026

pBBR-INP-8IJZ

This work is derived from pBBR1MCS-OmpA-mCherry and pUC19-8IJZ, and it has undergone codon optimization. This composite part combines OmpA(21.4kda) and 8IJZ, we succeeded in transferring this plasmid into Escherichia coli Nissle 1917 and let it express 8IJZ. The plasmid uses lac promotor and has kanamycin resistence.

Figure 1. Colony PCR results of pBBR-INP-8IJZ

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8IJZ is an anti-VEGF nanobody, and it has 2 chains.These two chains all show great affinity with VEGF, we plan to use pBBR1MCS-2 plasmid as a backbone and transfer 8IJZ into Escherichia coli Nissle 1917, and finally succeeded in expressing 8IJZ.

Figure 2. structure of 8IJZ， from PDB

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 4659
  Illegal XbaI site found at 3189
  Illegal PstI site found at 2016
  Illegal PstI site found at 3177
  Illegal PstI site found at 4427
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 4659
  Illegal PstI site found at 2016
  Illegal PstI site found at 3177
  Illegal PstI site found at 4427
  Illegal NotI site found at 1057
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 4659
  Illegal BglII site found at 1803
  Illegal BamHI site found at 3195
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 4659
  Illegal XbaI site found at 3189
  Illegal PstI site found at 2016
  Illegal PstI site found at 3177
  Illegal PstI site found at 4427
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 4659
  Illegal XbaI site found at 3189
  Illegal PstI site found at 2016
  Illegal PstI site found at 3177
  Illegal PstI site found at 4427
  Illegal NgoMIV site found at 2467
  Illegal NgoMIV site found at 2750
  Illegal NgoMIV site found at 3616
  Illegal NgoMIV site found at 5324
  Illegal AgeI site found at 5164
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1379
  Illegal SapI.rc site found at 2316
  Illegal SapI.rc site found at 2526