Things That we Have Done
From 2006.igem.org
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*Tet-forward | *Tet-forward | ||
* Tet-backwards | * Tet-backwards | ||
| + | * Cmr-backwards | ||
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| + | <font color="red">'''KanB'''</font color> | ||
| + | *Kan backwards is the only gene that we are completely positive has not been cloned into a plasmid. Further testing is needed to see if it has indeed been cloned. | ||
=RE and Hix= | =RE and Hix= | ||
| - | We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for part [http://partsregistry.org/Part: | + | We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for part Part BBa_J3101 [http://partsregistry.org/Part:BBa_J3101] for further details. |
| - | * As per the [http://2006.igem.org/Davidson_Assembly_Plan| assembly plan] we will be attaching the RE downstream of the double | + | |
| + | * As per the [http://2006.igem.org/Davidson_Assembly_Plan| assembly plan] we will be attaching the RE downstream of the double terminator BBa_B0015 [http://partsregistry.org/Part:BBa_B0015] | ||
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| + | *We also have two cultures with the correct sequences for Hix C. | ||
| + | *We have sent a "Master Plate" with cultures of all of the things that we have cloned to the Mammoths. | ||
| + | |||
| + | =Cloning Parts of the Constructs= | ||
| + | The underlined portion of each construct represents parts that have been successfully ligated and cloned: | ||
| + | |||
| + | pAra-RBS-Hix- <u>TetF-Hix</u>-<u>DFT-RE</u> | ||
| + | |||
| + | pAra-Hix-RBS-<u>TetF-Hix</u>-<u>DFT-RE</u> | ||
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| + | pAra-RBS-Hix-<u>TetB-Hix</u>-<u>DFT-RE</u> | ||
| + | |||
| + | pAra-Hix- TetB- SBR-Hix-<u>DFT-RE</u> | ||
| + | *SBR= backwards RBS | ||
Latest revision as of 13:44, 25 July 2006
Cloning
We have been able to successfully clone the following antibiotic pancakes
- Hin
- Hin with LVA degredation tag
- Cmr- forward
- Kan-forward
- Tet-forward
- Tet-backwards
- Cmr-backwards
KanB
- Kan backwards is the only gene that we are completely positive has not been cloned into a plasmid. Further testing is needed to see if it has indeed been cloned.
RE and Hix
We have been able to clone RE into cells. The RE that we are using has two point mutations but they should not compromise the activity of the Fis-binding sites please see the page for part Part BBa_J3101 [http://partsregistry.org/Part:BBa_J3101] for further details.
- As per the [http://2006.igem.org/Davidson_Assembly_Plan| assembly plan] we will be attaching the RE downstream of the double terminator BBa_B0015 [http://partsregistry.org/Part:BBa_B0015]
- We also have two cultures with the correct sequences for Hix C.
- We have sent a "Master Plate" with cultures of all of the things that we have cloned to the Mammoths.
Cloning Parts of the Constructs
The underlined portion of each construct represents parts that have been successfully ligated and cloned:
pAra-RBS-Hix- TetF-Hix-DFT-RE
pAra-Hix-RBS-TetF-Hix-DFT-RE
pAra-RBS-Hix-TetB-Hix-DFT-RE
pAra-Hix- TetB- SBR-Hix-DFT-RE
- SBR= backwards RBS
