Part:BBa_K4624609

accCD 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 final two subunits (C and D) of the acetyl-CoA carboxylase (accABCD), compatible with the 5' of the B4 position of the GoldenBraid 2.0 cloning grammar. This part is meant to be used with accCD B4 Reverse (BBa_K4624610) or accCD B4+B5 Reverse primer (BBa_K4624611) depending on the desired final syntax.

Below, we included the results of the gel electrophoresion of the PCR products. For the (1) accCD B4 part the primers BBa_K4624609 and BBa_K4624610 were used, while for (2) accCD B4+B5 part the primers BBa_K4624609 and BBa_K4624611 were used.

Figure 1: Gel electrophoresis of the PCR products for the (1) accCD B4 and (2) accCD B4+B5. Expected bands (bp): Lane 1: 2332 Lane 2: 2333, 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.