Part:BBa_K1122003

mTagBFP (BBa_K592100) under the control of lac promoter

The part is coding for the bright blue fluorescent protein mTagBFP (BBa_K592100) under the lac promoter. It is useful as a reporter inducible by IPTG.

Part construction

Part BBa_K1122003 was constructed in order to test Genabler assembly-compatible mTagBFP (BBa_K1122008) and one of the standard linkers Linker1. Since lacZ is known to be well expressed, Linker1 was designed to match lacZ upstream sequence containing an RBS and a start codon to enable protein expression.

A three-part Genabler assembly pAcceptor1(BBa_K1122010)-Plac-(Linker1)-mTagBFP(BBa_K1122008) was performed. Home-made DH5α competent E.coli cells were transformed with the assembly and plated on LB plates supplied with chloramphenicol and IPTG. Transformant colonies were brightly fluorescent under the UV light showing that the construct is functional (see Figure 1).

Figure 1. Cells that are blue on X-gal (left) are compared to DH5α competent cells with a 3-part assembly pAcceptor1-Plac-(Linker1)-mTagBFP (right) under the UV light.

References

Subach, O. M., I. S. Gundorov, et al. (2008). "Conversion of red fluorescent protein into a bright blue probe." Chem Biol 15(10): 1116-24.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]