DNA

Part:BBa_M1000

Designed by: Tom Knight   Group: UtahState_BE5930_-_S11   (2006-03-16)

Left mosaic end for Tn5 transpososome

  • Left end of a linear transposome sequence containing the 19 bp mosaic end in the forward orientation. The sequence also contains a PvuII cut site which releases the transpososome when cut with PvuII. Constructed compound parts intended for transposition must not contain other PvuII cut sites, or be PCR-amplified with blunt-end-producing enzymes (Pfu, Phusion) at the PvuII cut site between CAG and CTG.
  • This part, and the companion part M1001 provide BioBrick tools to construct Tn5 transposomes: linear DNA sequences which can be cut in vitro from a plasmid backbone, or created by PCR, combined in vitro with purified Tn5 transposase protein, electroportated into cells, and finally randomly insert into the bacterial chromosome.
  • The parts assembled between the two mosaic ends can be delivered, including antibiotic resistance markers or other functional chromosomal insertions.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


[edit]
Categories
//DNA/transposome/Tn5
//chassis/prokaryote/mflorum
Parameters
None