DNA

Part:BBa_K1732008

Designed by: Donna Lee   Group: iGEM15_Carnegie_Mellon   (2015-09-09)
Revision as of 09:10, 18 September 2015 by Donnalee (Talk | contribs)

pSB1C3-J23100-B0034-EGFP-His-B0015

pSB1C3-J23100-B0034-EGFP-His-B0015

His tagged eGFP and strong RBS. The J23100 strong constitutive promoter allows for high level expression of the fluorescent protein used to measure fluorescence.

A set of fluorescent proteins which includes BFP, GFP, eGFP, E0040, OFP, YFP, and RFP with a J23100 constitutive promoter and RFP with a J23115 constitutive promoter were characterized. The relative fluorescence as well as the signal-to-noise ratio of all the proteins in the MACH competent cell line were calculated. eGFP was measured at (ex/em = 488nm/509nm).


FIntensity.jpg

FQuantitation.jpg

Sequence alignment of original sequence to codon optimized sequence, indicating the differences in amino acids.

Alignment eGFP.jpg


The graphs below show the data for the original set of fluorescent proteins prior to being codon optimized via the IDT codon optimization tool. The set consists of His tagged RFP, OFP, YFP, eGFP, and BFP, each with a strong RBS and WT lac promoter that allows for Lac induction of fluorescence.

Fluorescent proteins characterization.jpg

Fluorescent proteins Amounts.jpg

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
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 212
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 77


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