DNA

Part:BBa_K1732007

Designed by: Donna Lee   Group: iGEM15_Carnegie_Mellon   (2015-09-09)
Revision as of 01:01, 19 September 2015 by Li.kenn (Talk | contribs)

pSB1C3-J23100-B0034-BFP-His-B0015

J23100-B0034-BFP-His-B0015

His tagged mTagBFP codon optimized for E.coli (BBa_K1732021) and strong RBS (BBa_B0034). The J23100 strong constitutive promoter (BBa_J23100) 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. With the exception of E0040 GFP, all the other sequences differ from the original because they were codon optimized for E.coli via IDT codon optimization tool. The relative fluorescence as well as the signal-to-noise ratio of all the proteins in the MACH competent cell line were calculated. BFP was measured at (ex/em = 399nm/456).


FIntensity.jpg

FQuantitation1.jpg


Sequence alignment of original sequence to codon optimized sequence.

Alignment BFP.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
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 285
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 607
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 518
    Illegal BsaI.rc site found at 683


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