Part:BBa_I50042:Design
From partsregistry.org
pSC101 origin of replication
- 10COMPATIBLE WITH RFC[10]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
Design Notes
The original design for BBa_I50042 was BBa_I50040. Synthesis of BBa_I50040 (designed pSC101 origin) was straightforward but testing revealed that BBa_I50040 was nonfunctional as a replication origin presumably due to introduced mutations.
In constructing BBa_I50042, the replication origin boundaries were chosen quite liberally (i.e. they might include more sequence than is strictly necessary) in part because BBa_I50040 was shown to be non-functional.
BBa_I50042 is very similar to the replication origin used in Bujard's pZS4Int-1 vector and in Michael Elowitz's repressilator. However, a mutation was introduced to eliminate a SpeI site for compatibility with the BioBrick assembly standard. BBa_I50042 is also the origin of replication in pSB4* series plasmids.
Source
We constructed BBa_I50042 by PCR using pSB4A3-P1010 as a template and amplification primers I50042-f (5'-GTTTCTTCGAATTCGCGGCCGCTTCTAGAGCTGTCAGACCAAGTTTACGAG-3') and I50042-r (5'-GTTTCTTCCTGCAGCGGCCGCTACTAGTAGTTACATTGTCGATCTGTTC-3').
References
- Cohen SN and Chang AC. Revised interpretation of the origin of the pSC101 plasmid. J Bacteriol 1977 Nov; 132(2) 734-7. pmid:334752.
- Lutz R and Bujard H. Independent and tight regulation of transcriptional units in Escherichia coli via the LacR/O, the TetR/O and AraC/I1-I2 regulatory elements. Nucleic Acids Res 1997 Mar 15; 25(6) 1203-10. pmid:9092630.
- Elowitz MB and Leibler S. A synthetic oscillatory network of transcriptional regulators. Nature 2000 Jan 20; 403(6767) 335-8. doi:10.1038/35002125 pmid:10659856.
