Difference between revisions of "Part:BBa K638402"

Revision as of 14:42, 20 September 2011

TorA tag variant

This TorA leader sequence variant has had been successfully used to export GFP to the periplasm of E.coli as described [http://www.ncbi.nlm.nih.gov/pubmed/11123687 here].

We ordered this sequence as a synthetic oligonucleotide in two halves: one 5-->3, one 3-->5, with 20bp overlap. We then joined these parts using PCR to make the full sequence. This is essentially a deliberate primer dimer. We then inserted this sequence into our previous constructs using [http://www.synbio.org.uk/gibson/downloads/files/RFC57.pdf Gibson assembly], an extremely powerful method of assembling DNA constructs that is compatible with Standard Assembly.

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_K638402 short</partinfo>
-We found a TorA sequence which had been successfully used to export GFP to the periplasm of E.coli.
+This TorA leader sequence variant has had been successfully used to export GFP to the periplasm of E.coli as described [http://www.ncbi.nlm.nih.gov/pubmed/11123687 here].
-The sequence was originally taken from this paper:
-Export of active green fluorescent protein to the periplasm by the twin-arginine translocase (Tat) pathway in Escherichia coli
-Joanna D. Thomas1, Richard A. Daniel2, Jeff Errington2, Colin Robinson1,*
-The sequence was added into our constructs as part of a gibson assembly primer.
+We ordered this sequence as a synthetic oligonucleotide in two halves: one 5-->3, one 3-->5, with 20bp overlap. We then joined these parts using PCR to make the full sequence. This is essentially a deliberate primer dimer. We then inserted this sequence into our previous constructs using [http://www.synbio.org.uk/gibson/downloads/files/RFC57.pdf Gibson assembly], an extremely powerful method of assembling DNA constructs that is compatible with Standard Assembly.
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