Part:BBa_K1321338
T7 Expression vector
This part enables easy addition of a promoter and RBS to any coding part which does not already have these parts. We used this part as an expression vector backbone for which to clone in any insert beginning with the start codon to allow us to actually express the protein. This part can be used to clone in both standard RFC10 Biobricks with the "N-part" prefix, or RFC25 Freiburg Fusion parts.
The part was designed so that the scar from assembly will leave an optimal 6 bp between the RBS and the start codon:
- This backbone part is cut with SpeI and PstI
- The desired insert part is cut with XbaI and PstI
- The resulting SpeI-XbaI scar is TACTAGA, where the last A is the first A of the subsequent ATG start codon.
We found it useful to clone this way since we could have a protein coding part on its own, which can easily be assmelbed into different backbones with different combinations of promoter or RBS. For example we also used part J04500 as an expression backbone in a similar manner.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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