Part:BBa_K2243032:Design
phiC31 attB_435R_phiC31 attP
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
A set of desirable unidirectional terminators were selected based on Nasri H, et. al Nature Methods 2013 and they were then constructed on the pSB4C5 vector. Terminator strength was tested using flow cytometry and corresponding Ts value was calculated.
Then attB/P sites were then introduced.
Two systems- the system containing sandwiched terminators and the system expressing corresponding recombinases were co-transformed into DH5alpha cells. Ts values of terminators sandwiched with attB/P sites were measured with flow cytometry.
Source
Streptomyces bacteria Phage phiC31
Terminators were commercially synthesized.
References
Dyakonov T, Muir A, Nasri H, et al. Characterization of 582 natural and synthetic terminators and quantification of their design constraints[J]. Nature Methods, 2013, 10(7):659-664.
Yang L, Voigt CA, et al. Permanent genetic memory with >1-byte capacity. Nat Methods. 2014 Dec; 11(12):1261-6.