Coding

Part:BBa_K2957001

Designed by: Cloning Team (Melody, Margaret, Krissy, Ethan)   Group: iGEM19_MIT   (2019-09-21)
Revision as of 00:57, 7 October 2019 by Mwu (Talk | contribs)


hEF1a, IL-8 (CXCL-8)

Contents: IL-8 (CXCL-8), FLAG Tag, with Carb 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 an 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 Flag tag attached.

How to use it?: You can use this part for Type IIS Cloning. It has BsaI cut sites that allow 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 pages on our MIT iGEM 2019 wiki: https://2019.igem.org/Team:MIT/Parts. It also has a Flag tag attached to it (which makes it different from pIG_048, BBa K779200) so this allows it to be tracked and then blotted with ELISA.

Sequence and Features


Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal XbaI site found at 3667
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2684
    Illegal BamHI site found at 3290
    Illegal BamHI site found at 3569
    Illegal XhoI site found at 3083
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal XbaI site found at 3667
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal XbaI site found at 3667
    Illegal PstI site found at 2430
    Illegal PstI site found at 2935
    Illegal NgoMIV site found at 197
    Illegal NgoMIV site found at 2818
    Illegal AgeI site found at 2025
    Illegal AgeI site found at 2196
    Illegal AgeI site found at 3673
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI site found at 2073
    Illegal SapI.rc site found at 9


[edit]
Categories
Parameters
None