Template:Team:UC Berkeley/Notebook/DV construction

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Contents


K112701

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

K112702

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-K112702
-----------------------------------------------
dv007   Forward biobricking of <His-Tag!     CATCATCATCATCATCATtaaGGATCC
dv008   Reverse biobriquing of <His-Tag!     GGATCCttaATGATGATGATGATGATG

K112703

Mix dv009/dv010 on pBca9145-Bca1117    (41bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+41, L)
Product is pBca1256-K112703
-----------------------------------------------
dv009	Forward Biobricking of His-Tag>
ccataGAATTCatgAGATCTatgCATCATCATCATCATCAT
dv010   Reverse Biobricking of His-Tag>
ATGATGATGATGATGATGcatAGATCTcatGAATTCtatgg

K112704

Mix dv011/dv012 on (template?)		(60 bp, EcoRI/BamHI/DpnI)
Sub into pBca1256			(EcoRI/BamHI)
Product is pBca1256-K112704
----------------------------------------
dv011		Forward Biobricking of <S-Tag!
GATCTAAAGAAACCGCTGCTGCTAAATTCGAACGCCAGCA
dv012		Reverse Biobricking of <S-Tag!
GGATCCttaCGCTGTCCATGTGCTGGCGTTCGAATTTAGC