Part:BBa_J107013

HSL inducible RiPS generator and ORF expresser

Overview:

This part is composed by BBa_F2620 HSL inducible device and a strong RBS (BBa_B0034) assembled downstream of the lux promoter (BBa_R0062). If this part is contained in a vector that has a transcriptional terminator downstream of the BioBrick Suffix (e.g. pSB1A3 or pSB1AK3 which have the E. coli his terminator BBa_B0053), an ORF can be quickly assembled and tested under the control of the lux HSL inducible promoter.

Usage:

The ORF can simply be a coding sequence (from ATG to a stop codon), without any RBS or transcriptional terminator. BBa_J107013 can be used as follows:

• digest the plasmid containing BBa_J107013 with SpeI-PstI;
• digest the ORF with XbaI-PstI;
• ligate the two DNA fragments and transform them in E. coli;
• let the transformed cells grow and induce them with 3OC₆HSL to trigger the expression of the ORF.

In B. Canton et al.(2008) (see References) and in BBa_F2620 Registry page the induction curve of BBa_F2620 is reported, so that users can choose the optimal HSL concentration. HSL between 100nM and 1uM should be sufficient for the ORF expression at very high levels.

Compatibility:

For the reasons explained above, this part must be used in vectors that have a transcriptional terminator downstream of the BioBrick Suffix (e.g. pSB1A3 or pSB1AK3). Otherwise, the ORF must be ligated to a transcriptional terminator (e.g. BBa_B0015) before being assembled with BBa_J107013.

This part should be compatible with all E. coli strains because the inducibility with HSL is genotype-independent.

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
  INCOMPATIBLE WITH RFC[1000]

  Illegal BsaI.rc site found at 1004