Coding

Part:BBa_K1413043

Designed by: Sophia Belkhelfa   Group: iGEM14_Evry   (2014-10-07)
Revision as of 18:09, 2 November 2014 by SophiaB (Talk | contribs) (Usage and Biology)

Transposase Tn10

This part is the coding sequence for the transposase Tn10. A cut-and-paste mechanism where the transposase allows insertion of a transposon randomly in the genome. The transposon must be surrounded by IS10 sequences.


Usage and Biology

This part is extract to plasmid pNK2 (see BBa_K1413044). This part is used to insert the transposon in genome of some bacteria like BL21 and Pseudovibrio denitrificans.


Picture of an electrophorese gel samples come from the PCR which amplify the kanamycineR cassette. All clones have the insertion of the transposon. The Control, Pseudovibrio denitrificans, do not the insertion as expected.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 244
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


References

-Bender, J., & Kleckner, N. (1988). Genetic Evidence That TnlO Transposes by a Nonreplicative Mechanism, 45, 801–815
-Crellin, P., & Chalmers, R. (2001). Protein±DNA contacts and conformational changes in the Tn 10 transpososome during assembly and activation for cleavage, 20(14).

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