Coding

Part:BBa_K2455005

Designed by: Jon Fugl   Group: iGEM17_UCopenhagen   (2017-10-27)


CPP-mTag BFP

This is an improved version of the Blue Fluorescent Protein (mTag BFP; BBa_K592100) carrying an N-terminal nona-arginin tag, and a C-terminal hexa-histidine tag. The R9-tag is a Cell-Penetrating Peptide giving its associated protein the ability to pass through plasma membranes.

For our study we demonstrated that this improved version of mTag BFP had fluorescence of similar levels to the original protein, and was able to stain both Escherichia coli and Pseudomonas putida cells, presumably through internalisation of the protein.

Previous studies have shown R9 associated proteins to be able to enter a variety of cell types (Chang et al., 2005).

DISCLAIMER: This biobrick carries an illegal EcoRI site partly in the end of the BFP and party in the beginning of the linker region - For data and design please see CPP-SYFP2.

Sequence and Features

Assembly Compatibility:
  • 10
    INCOMPATIBLE WITH RFC[10]
    Illegal EcoRI site found at 741
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal EcoRI site found at 741
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal EcoRI site found at 741
  • 23
    INCOMPATIBLE WITH RFC[23]
    Illegal EcoRI site found at 741
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal EcoRI site found at 741
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal EcoRI site found at 741


[edit]
Categories
//cds/reporter
//collections/chromoprotein/uppsala
//function/reporter/fluorescence
Parameters
abs399 nm
colorBlue
directionForward
emit456
excite399
functionCell-penetrating, Fluorescent
proteinmTag BFP
tagC-terminal hexa-histidine