DNA/Replication origins
The genetic element responsible for the replication of plasmids during cell growth and division is called a replication origin (also "origin of replication" or simply "origin"). There are several different replication origins and they differ in their plasmid copy number per cell (how many molecules of the plasmid are maintained in the cell), mechanism of copy number control, cell-to-cell copy number variation, and even the degree of coiling of the physical DNA. Thus, BioBrick® parts, devices and systems can operate very differently from one plasmid backbone to another.
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Several replication origins have been designed in the Registry. Although most of them are not available as individual BioBrick parts, some can be obtained by PCR of existing plasmid backbones in the Registry using appropriate primers. In fact, when making new BioBrick vectors using the BioBrick base vector, Reshma simply assembled a PCR-amplified linear DNA encoding the replication origins with the antibiotic resistance markers. (The PCR primers included the BioBrick prefix and suffix so that the linear DNA encoding the origin was ready for digestion with EcoRI and SpeI. |
| Name | Description | Replicon | Copy number | Chassis | Length |
|---|---|---|---|---|---|
| BBa_I50000 | F plasmid backbone with BioBrick sites removed. | 4640 | |||
| BBa_I50001 | F plasmid backbone with BioBrick sites removed, reverse | 4640 | |||
| BBa_I50022 | Minimal pUC19-derived high copy replication origin | 674 | |||
| BBa_I50030 | pBR322 replication origin | 2016 | |||
| BBa_I50032 | p15A replication origin | 1554 | |||
| BBa_I50040 | nonfunctional pSC101 origin of replication | 2226 | |||
| BBa_I50041 | pSC101 origin of replication, reverse | 2037 | |||
| BBa_I50042 | pSC101 origin of replication | 2037 | |||
| BBa_I50050 | R6K replication origin | 312 | |||
| BBa_I50052 | pSC101 temperature sensitive origin of replication | 2037 | |||
| BBa_J61001 | [R6K] Origin of replication | 406 | |||
| BBa_K112125 | OriV origin of replication | 646 | |||
| BBa_K125020 | lac promoter/rbs/rep/oriV/lac promoter/rbs/aadA/oriT | 4841 | |||
| BBa_K125340 | Origin of Vegetative Replication | 424 | |||
| BBa_K320000 | Asaia replication origin | 1391 | |||
| BBa_K320004 | Origin for Asaia and resistance to Kanamycin | 2374 | |||
| BBa_K524203 | pSC101 heat sensitive origin of replication with deletion in par locus | 1941 | |||
| BBa_K533000 | p15A replication origin | relaxed, high copy number | 914 | ||
| BBa_K864050 | pSC101 low copy origin of replication | 2216 | |||
| BBa_K864051 | pSC101ts temperature sensitive low copy origin of replication | 2216 |
References
<biblio>
- Chang-JBacteriol-1978 pmid=149110
- Cohen-JBacteriol-1977 pmid=334752
- Bolivar-Gene-1977 pmid=344137
- Bazaral-J-Mol-Biol-1968 pmid=4939624
- Hershfield-Proc-Natl-Acad-Sci-USA-1974 pmid=4610576
</biblio>