Part:BBa_K1943017:Design

Puro+T2A

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 481
  

Design Notes

This year we use mammalian cells as carriers. So in our plasmids, there are several kinds of anti-antibiotics genes which are used for screening. Usually, we will not use anti-antibiotics genes individually. We often use them together with some core protein such as GECO. However, it is monocistronic in the eukaryotic cells. There is a DNA sequence called 2A which connects two coding sequence and then both two call be expressed by the same promoter. We design primers and do PCR of this anti-antibiotics genes coding sequence together with 2A sequence and ligate it to pSB1C3 backbone.

Source

The plasmid was from our instructor, Huangwei's lab. And we design primers to copy down by PCR.

References

1. Parts Registry Assembly Help
2. Parts Registry Transformation Guideline