Difference between revisions of "Part:BBa P1016"

Revision as of 19:39, 9 July 2007

ccdB cassette without ccdA-

The CcdB protein, constitutively expressed by P1016, is lethal to most of the BioBrick cell strains, only DB3.1 is resistant.

This part is designed to be located in the multiple cloning site of low copy BioBrick vectors. BBa_P1016 provides selection against non-insert containing clones (see below).

P1016 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or religated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result.

A new set of low copy BioBrick plasmids are available with the BBa_I52001 insert (which includes BBa_P1016). See I52001 Physical DNA for the current list of plasmids that are available in this form.

Usage and Biology

For more information on how to use this brick, visit its Featured Parts:Cell Death page.

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
INCOMPATIBLE WITH RFC[1000]
Illegal BsaI site found at 323

@@ Line 12: / Line 12: @@
 This part is designed to be located in the multiple cloning site of low copy BioBrick vectors.  [[Part:BBa_P1016|BBa_P1016]] provides selection against non-insert containing clones (see below).
-P1016 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or religated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result.  I50020 is useful because it is a high copy origin located within the BioBricks cloning site.  Any plasmids containing this part will be propagated (and thus purified) at high copy.  Once the insert of interest is cloned into the plasmid, the I50020 part is eliminated and the plasmid returns to the copy number of the primary origin in the plasmid (usually low copy).
+P1016 is used when putting BioBrick parts into BioBrick plasmids. The part to be inserted and the plasmid are cut with BioBrick enzymes and mixed. The mixture will include both the original uncut or religated plasmid and the desired structure. However, because of CcdB, all of the cells containing the original plasmid die and the surviving colonies are the desired result.
 A new set of low copy BioBrick [[Help:Plasmids|plasmids]] are available with the [[Part:BBa_I52001|BBa_I52001]] insert (which includes BBa_P1016).  See [https://parts.igem.org/cgi/partsdb/dna.cgi?part_name=I52001 I52001 Physical DNA] for the current list of plasmids that are available in this form.<br>