Difference between revisions of "Part:BBa K838001"
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__NOTOC__ | __NOTOC__ | ||
<partinfo>BBa_K838001 short</partinfo> | <partinfo>BBa_K838001 short</partinfo> | ||
+ | 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. | |
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
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+ | '''1) Cloning:''' | ||
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+ | 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. | ||
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+ | 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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+ | [[File:Team_EPF_Lausanne_simpleswitch.png|500px]] | ||
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+ | '''2) Transfection''' | ||
+ | Co-transfect the combination of LovTAP-Vp16 and readout in mammalian cells! | ||
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Revision as of 17:40, 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.
Sequence and Features
Assembly Compatibility:
- 10INCOMPATIBLE WITH RFC[10]Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 424
Illegal BglII site found at 381 - 23INCOMPATIBLE WITH RFC[23]Illegal EcoRI site found at 424
Illegal SpeI site found at 295
Illegal SpeI site found at 303 - 25INCOMPATIBLE 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 - 1000COMPATIBLE WITH RFC[1000]