Difference between revisions of "Part:BBa K777125"

Revision as of 14:30, 20 September 2012

Constitutive J23100 promoter

This part is a combination of a constitutive Anderson promoter with the downstream RFP (for more information check out J61002 and J23100) and the plasmid backbone pSB1C3.

Usage and Biology

This construct can be useful in several ways, just like the original part J61002. However, if you plan to send your parts in, they are already in the correct backbone, pSB1C3.

You may use this plasmid to insert a promoter of your choice between the XbaI/EcoRI and SpeI sites. This will provide you with a downstream mRFP as a reporter. Look up the specifications for restriction sites here.
Furthermore, you can clone a gene of interest downstream of the constitutive Anderson promoters to express it at a desired rate. For this, the SpeI and PstI site can be used. Look up the specifications for restriction sites here.
It can provide a nice selection system for inserting your basic parts into the pSB1C3 vector via the EcoRI/XbaI and PstI site. If you use a construct containing a strong promoter e.g. K777125, colonies containing a self-ligated plasmid after transformation will turn red and you can select for the non-red colonies. Your basic parts can be easily inserted into the pSB1C3 vector and the RFP provides a nice selection system.

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 8
Illegal NheI site found at 31
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

Fig. 1: Plasmid map of K777125

@@ Line 5: / Line 5: @@
 ===Usage and Biology===
-To be continued...
+This construct can be useful in several ways, just like the original part [https://parts.igem.org/Part:BBa_J61002 J61002]. However, if you plan to send your parts in, they are already in the correct backbone, [https://parts.igem.org/Part:pSB1C3 pSB1C3].
+# You may use this plasmid to insert a promoter of your choice between the XbaI/EcoRI and SpeI sites. This will provide you with a downstream mRFP as a reporter. Look up the specifications for restriction sites [https://parts.igem.org/Image:PBca1020-r0040.jpg here].
+# Furthermore, you can clone a gene of interest downstream of the constitutive Anderson promoters to express it at a desired rate. For this, the SpeI and PstI site can be used. Look up the specifications for restriction sites [https://parts.igem.org/Image:PBca1020-r0040.jpg here].
+# It can provide a nice selection system for inserting your basic parts into the pSB1C3 vector via the EcoRI/XbaI and PstI site. If you use a construct containing a strong promoter e.g. K777125, colonies containing a self-ligated plasmid after transformation will turn red and you can select for the non-red colonies. Your basic parts can be easily inserted into the pSB1C3 vector and the RFP provides a nice selection system.
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