Part:BBa_K2971010
crtE and crtB in a single open reading frame
This part contains the sequences of crtE and crtB connected by a short linker sequence. The stop codon of each gene has been removed to express the genes in a single open reading frame. The enzymes encoded by the genes supply the precursor of carotenoids when expressed in Escherichia coli.
The composite part was made by fusing crtEB (BBa_K2971010) and crtI (BBa_K2971001) via Gibson cloning. The part was then transformed into competent cells of E. coli. The presence of the insert was screened with colony pcr (figure 2). The colony with the insert was analyzed by sequencing to check for potential mutations or frame-shifts.
Figure 2: Colony PCR of cells transformed with crtEB Only one colony (1) showed the presence of the correct insert. 1 was then further investigated.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1602
Illegal BamHI site found at 841 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 43
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