Device

Part:BBa_K3292001

Designed by: Abril Diosdado Hernandez, Gerardo David Diaz Renovado   Group: iGEM19_Tec-Monterrey   (2019-10-16)
Revision as of 02:17, 17 October 2019 by AbrilDH (Talk | contribs)


T7-LacO Promoter + strong RBS + sialidase + sfGFP + 6x his-tag + double terminator

This device uses Bba_K2406020, Bba_J34801 and Bba_0010 to regulate the expression of a super folding GFP (Bba_I746916). The main objective of creating this device is to facilitate the secretion and characterization of any protein intracellularly produced after being adding its nucleotide next to the sfGFP. In our case, we decided to add the coding sequence of sialidase from M. viridifaciens. The CDS of this enzymes comes from Micromonospora viridifaciens and was optimized for its expression in E. coli BL21 cells. Encoded by the gen ned A, this enzyme releases sialic acids (N-acetylneuraminic acid) moieties from glycoproteins and glycolipids for use as carbon and energy sources due to its hydrolytic activity.

Additionally, for the proper characterization of this part, the sequence of the of the sfGFP was added to the construct. e of the of the sfGFP was added to the construct.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 721
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 1986
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 646
    Illegal BsaI.rc site found at 510
    Illegal BsaI.rc site found at 1647
    Illegal SapI.rc site found at 2055


[edit]
Categories
Parameters
None