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Template:Team:UC Berkeley/Notebook/SC construction
From 2008.igem.org
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pBca1256-K112600
Construction of <AP-tag> basic part K112600 Wobble PCR SC001/SC002 (66 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910) Product is pBca1256-K112600 ---------------------------------------------------- SC001F Forward Biobricking of <AP-tag> cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag SC002R Reverse Biobricking of <AP-tag> ccagtGGATCCttcatgccattcaattttctgcgcttcaaaaatatcg
pBca1256-K112601
Construction of <AP-tag! basic part K112601 Wobble PCR SC001/SC003 (69 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910) Product is pBca1256-K112601 -------------------------------------------------- SC001 Forward Biobricking of <AP-tag! cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag SC003 Reverse Biobricking of <AP-tag! ccagtGGATCCttattcatgccattcaattttctgcgcttcaaaaatatcg
pBca1256-K112602
Construction of <FLAG-tag> basic part K112602 Wobble PCR SC004/SC005 (45 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910) Product is pBca1256-K112602 ------------------------------------------------------------- SC004 Forward Biobricking of <FLAG-tag> cgataGAATTCatgAGATCTgactacaaggatgacgacg SC005 Reverse Biobricking of <FLAG-tag> ccagtGGATCCcttgtcgtcgtcatccttgtagtc
pBca1256-K112603
Construction of <FLAG-tag! basic part K112603 Wobble PCR SC004/SC006 (48 bp, EcoRI/BamHI) Sub into pBca1256 (EcoRI/BamHI, 2227+ 910) Product is pBca1256-K112603 ------------------------------------------------------------- SC004 Forward Biobricking of <FLAG-tag! cgataGAATTCatgAGATCTgactacaaggatgacgacg SC006 Reverse Biobricking of <FLAG-tag! ccagtGGATCCttacttgtcgtcgtcatccttgtagtc
pBca1256-K112604
Construction of {rbs_barstar>} basic part K112604
PCR mea035/sc007 on Bacillus amyloliquefaciens gen. (320 bp, EcoRI/BamHI)
Sub into pBca1256 (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112604
-------------------------------
mea035 Fwd biobricking of {rbs_barstar>}
cgataGAATTCatgAGATCTcataagaaaggagccgcacatg
sc007 Reverse biobricking of {rbs_barstar>}
cgttaGGATCCagaaagtatgatggtgatgtcgc
pBca1256-K112403
Construction of {prepro>} of PhoA basic part K112403 (114 bp, EcoRI/BamHI)
CR cb10007/cb10008 on MG1655 (EcoRI/BamHI, 2472+910, L)
Sub into pBca1256
Product is pBca1256-K112403
-------------------------------------------------------------
cb10007-phoA-F Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTgtacatggagaaaataaaatgaaacaaagc
cb10008-phoA-R Reverse biobricking of {prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
pBca1256-K112404
Construction of {prepro>} of LamB basic part K112404 (132 bp, EcoRI/BamHI)
PCR cb10009/cb10010 on MG1655 (EcoRI/BamHI, 2472+910, L)
Sub into pBca1256
Product is pBca1256-K112404
-------------------------------------------------------------
cb10009-lamB-F Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTgaaaagcaatgactcaggagatag
cb10010-lamB-R Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
pBca1256-K112605
Construction of {a~prepro>} of PhoA basic part K112605 (100 bp, EcoRI/BamHI)
PCR sc008/cb10008-phoA-R (EcoRI/BamHI, 2472+910, L)
Sub into pBca1256
Product is pBca1256-K112605
------------------------------------------------------
sc008 Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtaaaAtgaaacaaagcactattgcac
cb10008-phoA-R Reverse biobricking of {a~prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
pBca1256-K112606
Construction of {a~prepro>} of LamB basic part K112606 (111 bp, EcoRI/BamHI)
PCR 009/cb10010-lamB-R (EcoRI/BamHI, 2472+910, L)
Sub into pBca1256
Product is pBca1256- K112606
--------------------------------------------------
sc009 Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtagaatgatgattactctgc
cb10010-lamB-R Reverse biobricking of {a~prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
pBca1256-K112607
Construction of {rbs_prepro>} of PhoA basic part K112607 (113 bp, EcoRI/BamHI)
PCR sc010/cb10008-PhoA-R (EcoRI/BamHI, 2472+910, L)
Sub into pBca1256
Product is pBca1256-K112607
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sc010 Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaag
cb10008-phoA-R Reverse biobricking of {rbs_prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
pBca1256-K112608
Construction of {rbs_prepro>} of LamB basic part K112608 (126 bp, EcoRI/BamHI)
PCR sc011/cb10010-lamB-R on E.coli K12 MG1655 (EcoRI/BamHI, 2472+910, L)
Sub into pBjh1601AK
Product is pBca1256-K112608
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sc011 Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTcaatgactcaggagatagaatgatgattac has little longer annealing region. If you use shorter, GC content goes up little. I would use this:
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
cb10010-lamB-R Reverse biobricking of {rbs_prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
