Composite

Part:BBa_K2722004

Designed by: Enikö Baligács   Group: iGEM18_Munich   (2018-10-09)


J23106 Promoter - mTurquoise - 380 Terminator

Transcription is promoted by the constitutive promoter obtained from the iGEM parts registry: BBa_J23106.


After the promoter there is a Plmj ribozyme. This cleaves itself and leaves the mRNA starting very near of the Start codon. This ribozyme was inserted to prevent context dependencies (influence of secondary mRNA structures on the expression level). The sequence of this part was generated to: BBa_K2722006


The cyan fluorescent protein is expressed under the ribosome binding site (RBS) based on Elowitz repressilator. RBS sequence is obtained from the iGEM parts registry: BBa_B0034.


The coding sequence of the mTQ protein can be founnd in the iGEM registry: BBa_K2722003


This part is terminated by an Eck terminator. The sequence was generated in the iGEM registry: Part: BBa_K2722007


The protein mTurquoise is a bright cyan fluorescent protein.

The protein molecular weight is 26.88 kilodaltons.

Excitation wavelength: 434 nm

Emission wavelength: 474 nm

Extinction coefficient: 30,000 1/Mcm

Quantum yield: 0.84

Brightness: 25.2

pKa value: 4,5

fluorescence lifetime: 3.7


This protein is very well suited for a fluorescence resonance energy transfer (FRET) donor.


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 301
  • 1000
    COMPATIBLE WITH RFC[1000]


The complete and correct sequence including spacers between the basic parts is:

gatctttacggctagctcagtcctaggtatagtgctagcGAAGCTTCAGTCATAAGTCTGGGCTAAGCCCACTGATGAGTCGCTGAAATGCGACGAAACTTATGACCTCTACAAATAATTTTGTTTAAAGTCAGTaaagaggagaaaTAGACATGGTTAGCAAGGGTGAAGAACTGTTAACCGGCGTCGTGCCGATTCTGGTTGAGCTGGATGGTGATGTCAACGGTCACAAGTTTAGCGTTAGCGGTGAGGGCGAGGGCGACGCCACCTACGGTAAATTGACCCTGAAGTTTATCTGCACGACCGGTAAGCTGCCGGTTCCGTGGCCGACCCTGGTGACGACTCTGTCGTGGGGCGTGCAATGTTTCGCGCGCTATCCGGATCACATGAAACAGCATGACTTCTTTAAGAGCGCGATGCCGGAAGGCTACGTTCAGGAACGTACGATCTTTTTCAAAGACGACGGTAACTATAAGACCCGCGCAGAAGTCAAGTTCGAGGGTGACACGCTGGTGAATCGTATTGAGCTGAAAGGTATTGACTTTAAAGAGGACGGTAACATCCTGGGTCACAAACTGGAGTATAATTACTTCAGCGACAATGTGTACATCACCGCTGATAAACAGAAAAACGGCATTAAAGCAAACTTCAAGATCCGTCACAATATTGAAGATGGCGGCGTGCAATTGGCCGATCACTATCAACAGAACACCCCGATTGGCGATGGTCCGGTCCTGCTGCCAGATAATCACTACTTGAGCACGCAATCCAAACTGTCCAAAGATCCGAACGAAAAACGTGACCACATGGTCCTGCTGGAATTTGTTACCGCGGCGGGTATCACGCTgggtatggacgaactgtacaagTAAttcagccaaaaaacttaagaccGCCGGTCTTGTCCACTACCTTGCAGTAATGCGGTGGACAGGATCGgcggttttcttttctcttctcTGG


[edit]
Categories
//awards/composite_part
//awards/part_collection
//cds
//cds/reporter
//cds/reporter/cfp
//classic/measurement/uncategorized
//function/reporter/fluorescence
Parameters
colorTurquoise
emission474 nm
excitation434 nm
familyCyan Fluorescent Proteins - CFP