Difference between revisions of "Part:BBa K838001"

Revision as of 17:43, 26 September 2012

LovTAP readout This part needs to be used with part BBa_K838000, which is LovTAP-VP16.

The part consists of a TRP repressor and DsRed. The LovTAP-Vp16 protein attaches to the this readout and promotes transcription of the DsRed mRNA.

1) Cloning:

First clone the part into a mammalian expression vector such as [http://products.invitrogen.com/ivgn/product/V79020?ICID=search-product pcDNA3.1(+)] or [http://products.invitrogen.com/ivgn/product/V04450 pCEP4]. These are the two main expression vectors we used.

Any promoters already present must be removed since we want only want expression drive by LovTAP-VP16 and not constitutive expression. The part must have a polyA tail at the end as well.

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2) Transfection Co-transfect the combination of LovTAP-Vp16 and readout in mammalian cells!

Sequence and Features

Assembly Compatibility:

10
INCOMPATIBLE WITH RFC[10]
Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303
12
INCOMPATIBLE WITH RFC[12]
Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303
21
INCOMPATIBLE WITH RFC[21]
Illegal EcoRI site found at 424
Illegal BglII site found at 381
23
INCOMPATIBLE WITH RFC[23]
Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303
25
INCOMPATIBLE WITH RFC[25]
Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303
Illegal NgoMIV site found at 7
1000
COMPATIBLE WITH RFC[1000]

@@ Line 4: / Line 4: @@
 The part consists of a TRP repressor and DsRed. The LovTAP-Vp16 protein attaches to the this readout and promotes transcription of the DsRed mRNA.
-<!-- Add more about the biology of this part here
-===Usage and Biology===
@@ Line 22: / Line 18: @@
 '''2) Transfection'''
 Co-transfect the combination of LovTAP-Vp16 and readout in mammalian cells!
+<!-- Add more about the biology of this part here
+===Usage and Biology===