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Template:Team:UC Berkeley/Notebook/DV construction
From 2008.igem.org
Construction of [<Phns>] Basic Part K112701
PCR dv001/dv004 on Phns (150 bp, gp = A) PCR dv003/dv006 on Phns (225 bp, gp = B) PCR dv005/dv002 on Phns (342 bp, gp = C)
PCR mv001/dv006 on A+B (355 bp, gp = D)
PCR dv001/dv002 on C+D (697bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+697) Product is pBca1256-K112701
dv001 Forward Biobricking of [<Phns>] CCATAgaattcATGagatctGAAATATAGCTGTGCCATCAGCC dv002 Reverse Biobricking of [<Phns>] CCATAggatccGCACGAAGAGTACGGATG
dv003 Removing the 1st EcoRI site in [<Phns>] (F) GCCAGGAATGTAAGGgATTCAAAATTGTTC dv004 Removing the 1st EcoRI site in [<Phns>] (R) GAACAATTTTGAATcCCTTACATTCCTGGC dv005 Removing the 2nd EcoRI site in [<Phns>] (F) gctaaagaGttcgccgaagtagc dv006 Removing the 2nd EcoRI site in [<Phns>] (R) gctacttcggcgaaCtctttagc
Construction of <His-Tag! Basic Part K112702
Mixing of dv007/dv008 (27bp, EcoRI/BamHI/DpnI) Sub into (vector?)pBca1256 (EcoRI/BamHI, 2472+27, L) Product is pBca1256-K112110
dv007 Forward biobricking of <His-Tag! CATCATCATCATCATCATtaaGGATCC dv008 Reverse biobriquing of <His-Tag! GGATCCttaATGATGATGATGATGATG
PCR dv009/dv010 on pBca9145-Bca1117 (1060bp, EcoRI/BamHI/DpnI)
Sub into pBca1256 (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112100
dv009 Forward Biobricking of His-Tag> ccataGAATTCatgAGATCTatgCATCATCATCATCATCAT dv010 Reverse Biobricking of His-Tag> ATGATGATGATGATGATGcatAGATCTcatGAATTCtatgg
Contents |
K112000
PCR bx1/bx2 on Bjh1301 (688 bp, EcoRI/BamHI/DpnI)
Sub into pBca1256 (EcoRI/BamHI)
Product is pBca1256-K112000
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bx1 Forward Biobricking of {T4 Holin!}
ccagtGAATTCatgAGATCTatggcagcacctagaatatc
bx2 Reverse Biobricking of {T4 Holin!}
cgttaGGATCCttatttagcccttcctaatattctggccgcc
