Part:BBa_J70027:Design
Mesoplasma florum plasmid origin
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 372
Design Notes
No native restriction sites. Added biobrick ends.
The gene for PE contains no CGG codons.
The primer names are for J70023 -- this is a shortened version
GTTTCTTCGAATTCGCGGCCGCTTCTAGAGtaattataaaaacaccttattttac,J00023-F2
GTTTCTTCCTGCAGCGGCCGCTACTAGTAttgggagtgcggggtttggggtgc,J00023-R
The start of J70027 matches the origin start in the pSci1 plasmid.
There appears to be a frame shift mutation at base 659, shifting the PE gene. This is likely fatal to replication. Re-PCR from the original S. citri genomic DNA sample and re-transform.
Source
Genbank AJ969070
PCR from S. citri genomic DNA, kind gift of J. Renaudin, using J70023-F2 and J70023-R primers.
References
Breton M, Duret S, Arricau-Bouvery N, Béven L, Renaudin J. Characterizing the replication and stability regions of Spiroplasma citri plasmids identifies a novel replication protein and expands the genetic toolbox for plant-pathogenic spiroplasmas. Microbiology. 2008 Oct;154(Pt 10):3232-44. PMID: 18832328
Saillard C, Carle P, Duret-Nurbel S, Henri R, Killiny N, Carrère S, Gouzy J, Bové JM, Renaudin J, Foissac X. The abundant extrachromosomal DNA content of the Spiroplasma citri GII3-3X genome. BMC Genomics. 2008 Apr 28;9:195. PMID: 18442384
Berho N, Duret S, Danet JL, Renaudin J. Plasmid pSci6 from Spiroplasma citri GII-3 confers insect transmissibility to the non-transmissible strain S. citri 44. Microbiology. 2006 Sep;152(Pt 9):2703-16. PMID: 16946265