Part:BBa_K2957009

hEF1a, NeonGreen-C5a

Contents: hEF1a, C5a, NeonGreen Tag, SynthpA, with Carb resistance

What is it?: A Composite part; it is a genetic circuit with hEF1a promoter and C5a gene allowing for constitutive C5a expression

What does it do?: It codes for the chemokine, C5a. It was important for our project in particular because of its involvement in the immune system and is known to create neutrophil swarming. It has a NeonGreen tag attached.

How to use it?: This part was created with Type IIS Cloning and transfected into HEKs. It also has a NeonGreen tag attached to the C5a so this allows us to visualize it and determine transfection efficiency. See more on the Parts Overview Page or MIT iGEM 2019 wiki: https://2019.igem.org/Team:MIT/Parts

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal EcoRI site found at 3971
  Illegal XbaI site found at 4018
  Illegal SpeI site found at 3083
  Illegal PstI site found at 464
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal EcoRI site found at 3971
  Illegal SpeI site found at 3083
  Illegal PstI site found at 464
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal EcoRI site found at 3971
  Illegal BglII site found at 1932
  Illegal BamHI site found at 480
  Illegal XhoI site found at 2048
  
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal EcoRI site found at 3971
  Illegal XbaI site found at 4018
  Illegal SpeI site found at 3083
  Illegal PstI site found at 464
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal EcoRI site found at 3971
  Illegal XbaI site found at 4018
  Illegal SpeI site found at 3083
  Illegal PstI site found at 464
  Illegal AgeI site found at 81
  Illegal AgeI site found at 4024
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 3611
  Illegal SapI.rc site found at 3584