Difference between revisions of "Part:BBa K103020:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=25_September_2008&arg1=29_September_2008&arg2=30_September_2008&arg3=9_October_2008&arg4=10_October_2008&arg5=13_October_2008&arg6=14_October_2008&name=Preparation%20of%20omega_linker%20under%20PT7%20%28BBa_K103020%29 | + | Preparation of BBa_K103019 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=25_September_2008&arg1=29_September_2008&arg2=30_September_2008&arg3=9_October_2008&arg4=10_October_2008&arg5=13_October_2008&arg6=14_October_2008&name=Preparation%20of%20omega_linker%20under%20PT7%20%28BBa_K103020%29 here] (entries from Univeristy of Warsaw 2008 iGEM team notebook). |
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+ | Omega_linker fragment was amplified from [http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha pACYC177+OmpA-omega-ΔA-alpha] using primers: 5' CAGCCATATGCTACTTACTCTAGCTTCCCGG 3' and 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 3'. Product was digested with NdeI and SacI and ligated into pET15b vector ([http://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega pET15b-OmpA-omega] with previously removed XbaI site, cutted with NdeI and SacI). | ||
+ | Omega_linker under control of lactose promoter was amplified using primers: 5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAA TTAATACG 3' and 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 3' | ||
+ | . PCR product were digested with EcoRI and BcuI and ligated into [https://parts.igem.org/wiki/index.php?title=Part:pSB2K3 pSB2K3] | ||
===Source=== | ===Source=== |
Revision as of 16:03, 28 October 2008
omega_linker under PT7
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 356
Design Notes
Preparation of BBa_K103019 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=7&arg0=25_September_2008&arg1=29_September_2008&arg2=30_September_2008&arg3=9_October_2008&arg4=10_October_2008&arg5=13_October_2008&arg6=14_October_2008&name=Preparation%20of%20omega_linker%20under%20PT7%20%28BBa_K103020%29 here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).
Omega_linker fragment was amplified from [http://2008.igem.org/Wiki/Team:Warsaw/vectors/pACYC177%2BompA-omega-A-alpha pACYC177+OmpA-omega-ΔA-alpha] using primers: 5' CAGCCATATGCTACTTACTCTAGCTTCCCGG 3' and 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 3'. Product was digested with NdeI and SacI and ligated into pET15b vector ([http://2008.igem.org/Wiki/Team:Warsaw/vectors/pET15b%2BOmpA-omega pET15b-OmpA-omega] with previously removed XbaI site, cutted with NdeI and SacI).
Omega_linker under control of lactose promoter was amplified using primers: 5' ATGAATTCGCGGCCGCTTCTAGAGATCCCGCGAAA TTAATACG 3' and 5' CCACTAGTACCGGATCCCGAACCACCCCCACCCCCGCTAC 3' . PCR product were digested with EcoRI and BcuI and ligated into pSB2K3