Difference between revisions of "Part:BBa K864012"

Revision as of 19:22, 27 September 2012

Low copy BioBrick standard vector with LacI

pSB4A15Iq(FRT) is a BioBrick standard vector with low copy pSC101 replication origin BBa_K864050 (~5 copies per cell) and ampicillin antibiotic resistance marker, especially usable for Lambda Red recombineering in E coli. The backbone sequence is based on pSB3T5, but the E coli His operon terminator BBa_B0053 has been replaced with the late terminator of the Salmonella phage P22.

This vector carries a LacIq cassette on the backbone for tight repression of any lac promoter cloned into the cloning site.

The resistance cassette is flanked by FRT sites (BBa_J61020) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase.

Usage and Biology

This part is available as pSB4A15-pUCori-redq. This facilitates high plasmid yields and fast red color expression on transformation plates.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4867
Illegal NheI site found at 2425
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NotI site found at 9
Illegal NotI site found at 4873
21
INCOMPATIBLE WITH RFC[21]
Plasmid lacks a prefix.
Plasmid lacks a suffix.
Illegal EcoRI site found at 4867
23
INCOMPATIBLE WITH RFC[23]
Illegal prefix found at 4867
Illegal suffix found at 2
25
INCOMPATIBLE WITH RFC[25]
Illegal prefix found at 4867
Plasmid lacks a suffix.
Illegal XbaI site found at 4882
Illegal SpeI site found at 2
Illegal PstI site found at 16
Illegal NgoMIV site found at 2319
1000
INCOMPATIBLE WITH RFC[1000]
Plasmid lacks a prefix.
Plasmid lacks a suffix.

@@ Line 8: / Line 8: @@
 The resistance cassette is flanked by FRT sites ([[Part:BBa_J61020|BBa_J61020]]) which allows for subsequent removal of the resistance cassette from the chromosome by the FLP recombinase.
-<!-- Add more about the biology of this part here
 ===Usage and Biology===
+This part is available as pSB4A15-[[Part:BBa_K864121|pUCori-redq]].  This facilitates high plasmid yields and fast red color expression on transformation plates.
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