Part:BBa_K2418001
Methylococcus capsulatus promoter - an improved version of BBa_K730001 - 1
Improved version of BBa_K730001 - 2
It was essential to find an appropriate promoter with high efficiency to enable the gene to work. Therefore, we looked for a promoter which can be found originally in Methylococcus capsulatus. The former iGEM team, iGEM12_juit found a nearly 1 000 base pairs long sequence which contained a promoter but they could not determine the exact location of the promoter. We did a search to find the 1 000 base pairs long sequence in Methylococcus capsulatus’ complete genome. This sequence contained not only the nucleotide sequence between two genes (the moxY and the mxaF) but did contain a partial part of each of the two genes. The putative moxY or mxaF used in this study did only contain the nucleotide sequence between the two genes, which must contain the promoter of either the moxY or the mxaF gene. Unfortunately, the orientation of the promoter is not known because the moxY and the mxaF genes are in different directions, therefore we could not determine neither the exact orientation of the promoter nor the exact sequence but we managed to approach the exact sequence of the promoter and apply it in such a way that the orientation was not needed to know.
The promoter was intended to ligate between the BglII restriction sites. This could enable the promoter to ligate in both orientations, randomly. The promoter was supplied with BglII restriction sites at both ends and addition AGTCAGTC nucleotides before and after each added AGTCAGTC nucleotides.
1. Addition AGTCAGTC bases
2. BglII restriction site
3. Promoter
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 239
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 9
Illegal BglII site found at 318 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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