Team:Freiburg Cloning Strategy

From 2008.igem.org

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<h2>'''Tabelle fuer Kathrin'''</h2>
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<h2>'''Cloning strategy'''</h2>
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Revision as of 20:22, 27 October 2008


Freiburg2008 small header.gif



Home

The Team

Project Report

Parts

Modeling

Notebook

Safety

CoLABoration

Cloning strategy

Step 1

Vector

digestion: EcoRI + PstI

Insert

digestion: EcoRI + PstI

 

BBa-J52017

_CMV-promotor

Step 2

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _ SPLIT-Linker

 

C-YFP

 

C-CFP

Step 3

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _egfR-Tm

 

_  N-β-Lactamase

 

_  C-β-Lactamase

 

_ SPLIT-Linker_ C-YFP

 

_ N-YFP

 

_ SPLIT-Linker_ C-CFP

 

_ N-CFP

 

_ BB058 (Luciferase)

 

_ BB057  (Luciferase)

Step 4

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _SP

 

_scFv-anti-NIP

 

_ Lipocalin

Step 5

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _SP_ scFv-anti-NIP

and

pMA-BBFR-+SP_ Lipocalin

_GGGS-linker (produced by Klenow fill in)

Step 6

Vector

digestion: AgeI+SpeI

Insert

digestion: NgoMIV+SpeI

 

pMA-BBFR _SP_ scFv-anti-NIP _ GGGS-Li

and

pMA-BBFR _ SP_ Lipocalin _

GGGS-Li

_ egfR-Tm _  N-β-Lactamase

 

_ egfR-Tm _  C-β-Lactamase

 

_ egfR-Tm _ SPLIT-Linker_ C-YFP

 

_ egfR-Tm _ N-YFP

 

_ egfR-Tm _ SPLIT-Linker_ C-CFP

 

_ egfR-Tm _ N-CFP

 

_ egfR-Tm _ BB058 (Luciferase)

 

_ egfR-Tm _ BB057  (Luciferase)

Step 7

Vector

digestion: SpeI + PstI

Insert

  digestion: XbaI + PstI

 

BBa-J52017_CMV

 

_SP_ scFv-anti-NIP_GGGS-Li_egfR-Tm_N-β-Lactamase

 

_ SP_ scFv-anti-NI _GGGS-Li_ egfR-Tm_C-β-Lactamase

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_SPLIT-Linker_C-YFP

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_N-YFP

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_SPLIT-Linker_C-CFP

 

_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_N-CFP

 

_ SP_ scFv-anti-NIP_GGGS-Li _ egfR-Tm_BB058 (Luciferase)

 

_ SP_ scFv-anti-NIP_GGGS-Li _ egfR-Tm_BB057  (Luciferase)

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-β-Lactamase

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_C-β-Lactamase

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_SPLIT-Linker_ C-YFP

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-YFP

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_SPLIT-Linker_ C-CFP

 

_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-CFP

 

_ SP_ Lipocalin _GGGS-Li__ egfR-Tm _ BB058 (Luciferase)

 

_ SP_ Lipocalin _GGGS-Li__ egfR-Tm _ BB057  (Luciferase)

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Retrieved from "http://2008.igem.org/Team:Freiburg_Cloning_Strategy"