![](https://parts.igem.org/images/partbypart/icon_plasmid_backbone.png)
Part:BBa_K4727009
Low copy backbone for K. pneumoniae and A. baumannii
This plasmid backbone was developed by Wang et al [1], it is provided of two ORIs: the first one a p15a medium/low copy number ORI for E. coli, the second one is capable o maintaining the plasmid both in Acinetobacter baumannii and Klebsiella pneumoniae
This backbone has been made standard RFC[10] compatible and provided with the RFC[10] prefix and suffix. Flanked by prefix and suffix is located part BBa_J23119 that has been demonstrated able to promote mRNA synthesis in K. pneumoniae and A. baumannii
This part is the result of and engineering cycle applied on part BBa_K4727000 out of the necessity to express a dCas expression cassette. Our experiments, and the literature, suggests that plasmid backbone BBa_K4727000 has an ORI for E. coli that provides a high number of copies (500-700). This information, together with the knowledge that the Cas protein if present in large quantities results in cellular toxicity, made us hypothesize that cloning a dCas expression cassette in a high copy number plasmid as BBa_K4727000 can lead to no results. So, out of this necessity this new part was born. It maintains the same ORI for
Sequence and Features
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found at 4850
Illegal suffix found at 4907 - 12INCOMPATIBLE WITH RFC[12]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4850
Illegal NheI site found at 2155
Illegal NheI site found at 4878
Illegal NheI site found at 4901
Illegal SpeI site found at 4908
Illegal PstI site found at 4922
Illegal NotI site found at 4856
Illegal NotI site found at 4915 - 21INCOMPATIBLE WITH RFC[21]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4850
Illegal BglII site found at 2348
Illegal XhoI site found at 2961
Illegal XhoI site found at 4966 - 23INCOMPATIBLE WITH RFC[23]Illegal prefix found at 4850
Illegal suffix found at 4908 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found at 4850
Plasmid lacks a suffix.
Illegal XbaI site found at 4865
Illegal SpeI site found at 4908
Illegal PstI site found at 4922
Illegal AgeI site found at 5462
Illegal AgeI site found at 5785 - 1000INCOMPATIBLE WITH RFC[1000]Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal SapI site found at 5194
None |