Composite

Part:BBa_K2921400

Designed by: Allison Kuo   Group: iGEM19_TAS_Taipei   (2019-10-14)
Revision as of 08:07, 18 October 2019 by Allishannn (Talk | contribs)


Strong Promoter + RBS + MBP + Double Terminator

This construct constitutively expresses a colored metal-binding protein MBP (Basic part: BBa_K346004) MBP is a lead binding peptide that can accumulate lead ions from aquatic environment.

Construct design (insert photo) We inserted an upstream constitutive strong promoter-RBS combination (Basic part: BBa_K880005) and a downstream strong double terminator (Basic part: BBa_B0015) so as to maximize plasmid and protein expression.

PCR


PCR check results, using the primers VF2 and VR, and sequencing results by Tri-I Biotech confirm this construct. (insert photo)

Characterization

To verify MBP expression in E. coli, we ran a SDS-PAGE gel, expecting a protein band at ___kDa. We prepared and lysed overnight liquid cultures of cells expressing MBP-mRFP, MBP only, and mRFP only. The SDS-PAGE reveals a darker band at ___ kDa as compared to the MBP-only and mRFP-only controls. (insert photo)


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 7
    Illegal NheI site found at 30
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


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Categories
Parameters
None