Part:BBa_K3440015

RhlI-AiiA

RhlI(BBa_C0070)-RBS(BBa_B0034)-AiiA(BBa_C0060)

Usage

This is an intermediate part used in the construction of BBa_K3440020 and BBa_K3440021

Characterisations

Due to the pandemics, we haven’t been able to use biobricks to create the iGEM Stockholm 2020 parts. Those parts were ordered as gene blocks from Integrated DNA Technologies Inc.. As a result, the sequences of the biobricks used are the same, but the scars between biobricks might differ, as well as the final size of the part.

After heat shock transformation of the pSB1C3 plasmid containing the BBa_K3440015, we picked colonies from plates (Figure 1) and PCR amplified them with primers VF and VR2.

Figure 1: plate of BBa_K3440015

We ran gels of the product at 180V and for 30 mins (Figure 2). We obtained the expected size for the bands (1779bp) for H5, H5, H8, H11 and H12.

Figure 2: Colony PCR Gel for BBa_K3440015 (H)

We therefore prepared plasmid preparations and glycerol stocks of those and sent some of them for sequencing to Microsynth AG. The sequence obtained corresponded to the expected part for H5 and H11.

No further characterisation was done since this was only an intermediate part used to build others.

Sequence and Features

Assembly Compatibility:

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

  Illegal BglII site found at 661
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 792
  
• 1000
  COMPATIBLE WITH RFC[1000]