Part:BBa_I739005

Promoterless expression cassette for EYFP +LVA (B0034.E0032.B0015)

Part Structure

The Biobrick encodes LVA-tagged EYFP (BBa_E0032) downstream of the ribosome binding site BBa_B0034. The transcription of EYFP is terminated by the double terminator BBa_B0015. This construct can be used as second cistron expressing a reporter gene in a multicistronic construct.

WARNING: This synthetically synthesized part is identical to part BBa_E0432. However, since BBa_E0432 is not properly working (probably because of the LVA tag of EYFP) , it should be retested. If there should still be issues with this part, one should switch to BBa_E0430 or BBa_E0434. Because part BBa_E0438 seems to be difficult to handle too, on could conclude that the LVA-tagged EYFP (BBa_E0032) is not working at all! So please be careful with those parts...

Mode of Action

Enhanced yellow fluorescent proteins (EYFP) are fluorochromes, commonly used as reporter proteins.

Purpose

This Biobrick was designed for the [http://parts.mit.edu/igem07/index.php/ETHZ ETHZ iGEM 2007 project] and initially belonged to the reporting part of the system. In the project description, this part was also termed Part 5. Together with Part 4, this Biobrick was thought to form the composite part BBa_I739015.

HOWEVER: Because of the above mentioned problems, this part was NOT USED. Instead it was replaced by BBa_E0430 which is termed Part 5new.

Testing

Checked for uniqueness of restriction enzyme cleavage sites:
Eco: ok
Xba: ok
Spe: ok
Pst: ok

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]