Device

Part:BBa_K3292001

Designed by: Abril Diosdado Hernandez, Gerardo David Diaz Renovado   Group: iGEM19_Tec-Monterrey   (2019-10-16)
Revision as of 01:41, 22 October 2019 by DamEsp (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 adding its nucleotide sequence next to the sfGFP.

Characterization

The characterization of this part included a Bradford assay to quantify the amount of protein present in the medium for each of the proposed constructs. The conditions for the assay are listed below.

The other characterization assay was a fluorescent measurement, where bacteria were growth at 30ºC for 3 hrs. Then 100µl of the grown bacteria were induced with IPTG at 0, 40, 200 and 400 mMm microtiter florescent measures were taken every half an hour for a period of 3 hours. Absorption and emission values were 395/509 nm.

800px-T--Tec-Monterrey--demostrate-grafica.png

Figure 1. sfGFP grpah, fluorescence vs time induced at different concentrations.

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