Part:BBa_K2387048

Split sfGFP controlled by inducible araC/pBAD promoter

This a split version of BBa_K2387047. The two halves of sfGFP are fused to leucine zippers to induce the reassembly. Both parts are under the control of the araC/pBad promoter and each part is under the control of a stron RBS. The reassembly process is irreversible. This part can be used as a positive control for experiments using BiFC, as well as a source for the fragments of sfGFP.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal NheI site found at 1205
  
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1144
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 979
  
• 1000
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI site found at 2126
  Illegal SapI site found at 961
  Illegal SapI.rc site found at 1251
  

Functional Parameters

In order to test the functionality of this split protein under the araC/pBad promoter, the absorbance and the fluorescence spectra of the full length protein and the split protein were measured after induction with 0.002% arabinose (Figure 1). The stability of the shape in both spectra for the full length protein and the split protein show that the split has been successful.

Figure 1: Absorbance and fluorescence spectra of full length sfGFP (left) and split sfGFP (right), measured after induction with 0.002% arabinose.

This protein is part of a collection of split proteins developed for BiFC analysis and tested with the CpxR system (BBa_K2387032). These proteins were characterized through multiple experiments using as interacting proteins antiparallel synthetic leucine zippers. The proteins of this collection are the ones found in Table 1.

Table 1: Split Proteins.
Protein	Part Number (Full length)	Part Number (Split Protein)
mRFP	BBa_K2387054	BBa_K2387055
eYFP	BBa_K2387003	BBa_K2387065
mVenus	BBa_K2387045	BBa_K2387046
sfGFP	BBa_K2387047	BBa_K2387048
mCerulean	BBa_K2387052	BBa_K2387053