Part:BBa_I52001:Design
From partsregistry.org
ccdB and nonfunctional pUC19 derived high copy origin
- 10COMPATIBLE WITH RFC[10]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 435
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
Design Notes
The BioBrick standard vectors are designed to be easy to use for their most common purpose: assembly of BioBrick standard biological parts. To meet this requirement, we included BBa_P1016 within the BioBrick cloning site. BBa_P1016 encodes the positive selection marker ccdB. Positive selection markers prevent one of the most common problems during assembly of BioBrick parts: contamination of the ligation reaction with uncut plasmid DNA[1]. Any cells transformed with the uncut plasmid DNA produce the lethal protein ccdB and die[2, 3, 4]. Note, however, that the drawback of this solution is that inclusion of this part requires that users propagate both the base vector and any derived vectors in E. coli strains tolerant of ccdB expression such as DB3.1[5, 6].
Source
BBa_P1016 (ccdB positive selection marker) and BBa_I50020 (nonfunctional high copy origin derived from pUC19).
References
- Bernard P, Gabant P, Bahassi EM, Couturier M, Université Libre de Bruxelles. Positive-selection vectors using the F plasmid ccdB killer gene." Gene 1994 Oct 11;148(1):71-4. pmid:7926841. Pubmed
- Bernard P, Gabant P, Université Libre de Bruxelles. Cloning and/or sequencing vector US Patent Number 5,910,438, 1999. Google Patents
- Bernard P, Gabant P, Université Libre de Bruxelles. Cloning and/or sequencing vector US Patent Number 6,180,407 B1, 2001. Google Patents
- Bernard P, Gabant P, Université Libre de Bruxelles. Cloning and/or sequencing vector US Patent Number 7,176,029 B2, 2007. Google Patents
- Bahassi, et al., Université Libre de Bruxelles. Interactions of CcdB with DNA gyrase. Inactivation of Gyra, poisoning of the gyrase-DNA complex, and the antidote action of CcdA., J. Biol. Chem. 1999 274: 10936-10944. pmid:10196173. Pubmed
- Bernard P. New ccdB positive-selection cloning vectors with kanamycin or chloramphenicol selectable markers. Gene 1995 Aug 30; 162(1) 159-60. pmid:7557407.
- Bernard P. Positive selection of recombinant DNA by CcdB. Biotechniques 1996 Aug; 21(2) 320-3. pmid:8862819.
- Bernard P and Couturier M. Cell killing by the F plasmid CcdB protein involves poisoning of DNA-topoisomerase II complexes. J Mol Biol 1992 Aug 5; 226(3) 735-45. pmid:1324324.
- Miki T, Park JA, Nagao K, Murayama N, and Horiuchi T. Control of segregation of chromosomal DNA by sex factor F in Escherichia coli. Mutants of DNA gyrase subunit A suppress letD (ccdB) product growth inhibition. J Mol Biol 1992 May 5; 225(1) 39-52. pmid:1316444.
