Regulatory

Part:BBa_K1675020:Design

Designed by: Jing Yang   Group: iGEM15_BIT-China   (2015-09-15)


P-atp2 mutant 226-B0034-lacz alpha


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
    COMPATIBLE WITH RFC[1000]


Design Notes

There is an EcoRI site exists in the P-atp2 sequence, so we mutate it to change it.


Source

Error prone PCR of P-atp2, then overlap extension of synthetic oligonucleotides.

The natural P-atp2 is from Corynebacterium glutamicum.

Lacz alpha is from https://parts.igem.org/wiki/index.php?title=Part:BBa_I732006


References

［1］ Le Y, Chen H, Zagursky R, et al. Thermostable DNA ligase-mediated PCR production of circular plasmid (PPCP) and its application in directed evolution via in situ error-prone PCR[J]. DNA research, 2013: dst016.

［2］ Pai J C, Entzminger K C, Maynard J A. Restriction enzyme-free construction of random gene mutagenesis libraries in Escherichia coli[J]. Analytical biochemistry, 2012, 421(2): 640-648.