"
Team:Freiburg Cloning Strategy
From 2008.igem.org
Tabelle fuer Kathrin
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Step 1 |
Vector digestion: EcoRI + PstI |
Insert digestion: EcoRI + PstI |
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BBa-J52017 |
_CMV-promotor |
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Step 2 |
Vector digestion: AgeI+SpeI |
Insert digestion: NgoMIV+SpeI |
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pMA-BBFR _ SPLIT-Linker
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C-YFP |
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C-CFP |
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Step 3 |
Vector digestion: AgeI+SpeI |
Insert digestion: NgoMIV+SpeI |
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pMA-BBFR _egfR-Tm
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_ N-β-Lactamase |
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_ C-β-Lactamase |
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_ SPLIT-Linker_ C-YFP |
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_ N-YFP |
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_ SPLIT-Linker_ C-CFP |
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_ N-CFP |
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_ BB058 (Luciferase) |
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_ BB057 (Luciferase) |
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Step 4 |
Vector digestion: AgeI+SpeI |
Insert digestion: NgoMIV+SpeI |
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pMA-BBFR _SP
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_scFv-anti-NIP |
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_ Lipocalin |
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Step 5 |
Vector digestion: AgeI+SpeI |
Insert digestion: NgoMIV+SpeI |
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pMA-BBFR _SP_ scFv-anti-NIP and pMA-BBFR-+SP_ Lipocalin |
_GGGS-linker (produced by Klenow fill in) |
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Step 6 |
Vector digestion: AgeI+SpeI |
Insert digestion: NgoMIV+SpeI |
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pMA-BBFR _SP_ scFv-anti-NIP _ GGGS-Li and pMA-BBFR _ SP_ Lipocalin _ GGGS-Li |
_ egfR-Tm _ N-β-Lactamase |
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_ egfR-Tm _ C-β-Lactamase |
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_ egfR-Tm _ SPLIT-Linker_ C-YFP |
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_ egfR-Tm _ N-YFP |
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_ egfR-Tm _ SPLIT-Linker_ C-CFP |
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_ egfR-Tm _ N-CFP |
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_ egfR-Tm _ BB058 (Luciferase) |
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_ egfR-Tm _ BB057 (Luciferase) |
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Step 7 |
Vector digestion: SpeI + PstI |
Insert digestion: XbaI + PstI |
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BBa-J52017_CMV
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_SP_ scFv-anti-NIP_GGGS-Li_egfR-Tm_N-β-Lactamase |
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_ SP_ scFv-anti-NI _GGGS-Li_ egfR-Tm_C-β-Lactamase |
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_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_SPLIT-Linker_C-YFP |
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_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_N-YFP |
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_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_SPLIT-Linker_C-CFP |
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_ SP_ scFv-anti-NIP_GGGS-Li_ egfR-Tm_N-CFP |
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_ SP_ scFv-anti-NIP_GGGS-Li _ egfR-Tm_BB058 (Luciferase) |
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_ SP_ scFv-anti-NIP_GGGS-Li _ egfR-Tm_BB057 (Luciferase) |
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_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-β-Lactamase |
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_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_C-β-Lactamase |
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_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_SPLIT-Linker_ C-YFP |
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_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-YFP |
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_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_SPLIT-Linker_ C-CFP |
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_ SP_ Lipocalin _GGGS-Li_ egfR-Tm_N-CFP |
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_ SP_ Lipocalin _GGGS-Li__ egfR-Tm _ BB058 (Luciferase) |
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_ SP_ Lipocalin _GGGS-Li__ egfR-Tm _ BB057 (Luciferase) |
