Template:Team:UC Berkeley/Notebook/DV construction

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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 (41 bp)
dv010   Reverse Biobricking of His-Tag>
ATGATGATGATGATGATGcatAGATCTcatGAATTCtatgg (41 bp)

K112704

Wobble PCR of dv011/dv012 (no 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

K112705

Mix dv013/dv014 (no template)           (47 bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+47, L)
Product is pBca1256-K112705
-----------------------------------------------
dv013	Forward Biobricking of S-Tag>
CCATAgaattcATGAGATCTatgGATCTAAAGAAACCGCTGCTGCTA (47 bp)
dv014   Reverse Biobricking of S-Tag>
CGCTGTCCATGTGCTGGCGTTCGAATTTAGCAGCAGCGGTTTCTTTA (47 bp)

K112706

PCR of dv015/dv016 (template?) (500bp, EcoRI/BamHI/DpnI) Sub into pBca1256 (EcoRI/BamHI, 2472+ 500, L) Product is pBca1256-K112706

dv015 Forward biobricking of <Pspv2> CCATAgaattcATGAGATCTctagaccttatgcagcgtaaggg dv016 Reverse biobricking of <Pspv2> GGATCCtGATAATGTtTGCAGGGGAATTATTTTGTATTGCC </pre>

K112707

PCR of dv017/dv018 (template?)         (499bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                       (EcoRI/BamHI, 2472+499, L)
Product is pBca1256-K112707
-----------------------------------------------
dv017 Forward biobricking of <Pspv>
CACTGgaattcatgAGATCTccttatgcagcgtaagggccgcaac
dv018 Reverse biobricking of <Pspv>
CGTACggatccGATAATGTtTGCAGGGGAATT
<pre/>