Part:BBa_K4235027
Tn7L: Left end of the Tn7 transposon segment
This is the left end of our transposon segment, required for the action of the enzyme transposase. Tn7 are terminal inverted transposon elements that flank both sides of the mini-attTn7 segment. These are used for site-specific transposition of heterologous gene circuits into attTn7 sites on a different chromosome and are more efficient in generating recombinant progeny than transposition through homologous recombination. Our project aimed at utilizing the mini-attTn7 segment for site specific transposition of our insert BBa_K4235000 from the pFastBac transfer vector onto the baculovirus genome (Bacmid), which is propagated in E. coli DH10Bac cells. Our mini-attTn7 circuit contains a Gentamicin Resistance gene BBa_K4235003, a multiple cloning site driven by a polyhedrin promoter BBa_K4235001, SV40 polyA signal and Tn7-L and Tn7-R ends for transposase activity. The Bac-to-Bac expression system exploits this mechanism of transposition to generate recombinant Bacmid particles in E coli DH10Bac, which can be isolated and used to infect insect cells for the production of recombinant proteins.
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]
//dna/transposon
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