Difference between revisions of "Part:BBa M1000"

Latest revision as of 20:38, 12 April 2007

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]

@@ Line 1: / Line 1: @@
 __NOTOC__
 <partinfo>BBa_M1000 short</partinfo>
-Left end of a linear transpososome 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 end, or be PCR amplified with blunt end producing enzymes (Pfu, Phusion) at the PvuII cut site between CAG and CTG.
+*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 transpososomes: linear DNA sequences which can be cut in vitro from a plasmid backbone, or created by PCR, combined in vitro with purified Tn5 trasposase protein, electroportated into cells, and finally randomly inserting into the bacterial chromosome.
+*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.
+*The parts assembled between the two mosaic ends can be delivered, including antibiotic resistance markers or other  functional chromosomal insertions.