Part:BBa_K103017:Design
OmpA_linker_alpha_linker under Plac
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1321
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
Preparation of BBa_K103016 is in details described [http://2008.igem.org/wiki/index.php?title=Team:Warsaw/JSTest&num=5&arg0=10_October_2008&arg1=13_October_2008&arg2=14_October_2008&arg3=15_October_2008&arg4=16_October_2008&name=Preparation%20of%20OmpA_linker_alpha_linker%20under%20Plac%20(BBa_K103017) here] (entries from Univeristy of Warsaw 2008 iGEM team notebook).
Alpha 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' GG GAGCTCGCTACTTACTCTAGCTTCCCGG 3' and 5' TCTGGAGGTGGAGGTAGCGG GGGTGGGGGTGGTTCGGGTGGAGGTGGT AAAACCGCGGCTCTTGCGC 3'.
Resulting product was used as a template for PCR with primers: 5' GG GAGCTCGCTACTTACTCTAGCTTCCCGG 3' and 5' ATACTAGTACCGGATCCAGAACCTCCTCCACCGCTACCTCCACCTCCAGAAC 3' (to obtain alpha_linker fragment), followed by digestion with SacI and BcuI.
Alpha_linker fragment was ligated into pSB2K3 vector containing lactose promoter and OmpA_linker (obtained by digest of pSB2K3 carrying BBa_K103018 with SacI and BcuI)