Part:BBa_K3028001
С-terminal part of beta-lactamase fused with dCas9 and double terminator
С-terminal part of beta-lactamase fused with dCas9 and double terminator
Part BBa_K1689013 is a С-terminal fragment of β-lactamase fused with dCas9.
BBa_K3219000: dCas9 enzyme is also known as a catalytically dead Cas9 enzyme[1]. It lacks traditional endonuclease activity (it does not cleave DNA) but preserves its ability to bind targets very specifically. This part was modified by IGEM_Peking team for DNA detection purposes. β-Lactamases - bacterial enzymes that can hydrolyze the amide bond of the β-lactam ring, hence inactivate β-lactam antibiotics (AmpR). Split β-Lactamase (196|198) [2] is used as a reporter, for example, to study different protein-protein interactions. When fused to dCas9, it can report the presence of a specific target by regaining its enzymatic activity.
Our part is a composition of the most commonly used terminator and BBa_K1689014. It was necessary to make such an improvement, because of all side effects of Cas synthesis for cells.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 1150
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 4
Illegal BamHI site found at 3429 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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