Coding

Part:BBa_K2957002

Designed by: Cloning Team (Melody, Margaret, Krissy, Ethan)   Group: iGEM19_MIT   (2019-09-21)


IL-8 (CXCL-8) with NeonGreen tag

Contents: IL-8 (CXCL-8), NeonGreen tag, with Spec resistance

What is it?: A Basic part; in this particular case we call it a pL0-G (through Modular Cloning Golden Gate Type II Assembly)

What does it do?: It codes for the chemokine, IL-8 (CXCL8), which is involved in inflammatory response. 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?: You can use this part for Type IIS Cloning. It has BsaI cut sites that allows for this. With other Basic parts like pL0-P (promoter/regulatory sites), pL0-T (terminator part), pL0-5.2/pL0-3 (Inert pieces that form the UTR regions around the genetic circuit), you can use this part to make a composite part. So the pL0's are placed in a given backbone depending on the kind of basic part it is. In this case, we have the CDS/gene part which is in a pL0-G backbone. See more on the Parts Overview Page or MIT iGEM 2019 wiki: <a href=https://2019.igem.org/Team:MIT/Parts>. It also has a NeonGreen tag attached to it (which makes it different from pIG_047, BBa K779200) so this allows us to visualize it and determine transfection efficiency.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal PstI site found at 460
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 460
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 234
    Illegal BamHI site found at 476
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal PstI site found at 460
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal PstI site found at 460
  • 1000
    COMPATIBLE WITH RFC[1000]


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Parameters
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