Part:BBa_K4624612

Laccase B4+(B5) Forward primer for GoldenBraid 2.0 cloning.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal XhoI site found at 12
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Usage

This part was designed for the addition of 5' appropriate overhangs to the lcc2 laccase where the signal peptide has been removed (BBa_K4624004). This part is meant to be used with Laccase B4 Reverse primer (BBa_K4624613) or Laccase B4+B5 Reverse primer (BBa_K4624614) depending on the desired final syntax.

The resulting part will compatible with the B4 or B4+B5 position of the GoldenBraid 2.0 cloning grammar, depending on the reverse primer used.

Below, we included the results of the gel electrophoresis of the PCR products. For the (7) laccase B4 the primers BBa_K4624612 and BBa_K4624613 were used, while for (8) laccase B4+B5. the primers BBa_K4624612 and BBa_K4624614 were used.

Figure 1: Gel electrophoresis of the PCR products for the (7) laccase B4 and (8) laccase B4+B5. Expected bands (bp): Lane 7: 1537 Lane 8: 1538, L=1kb plus DNA ladder

The successful addition of the appropriate 3' and 5' overhangs was confirmed upon the succesful insertion of the two parts into the universal part domesticator plasmid pUPD2, which is compatible only with domesticated parts, such as our PCR products.