Template:Team:UC Berkeley/Notebook/MJV construction

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Revision as of 02:18, 5 June 2008

Construction of {<Cre!} Basic Part K112100

PCR mv001/mv002 on pBca9145-Bca1117    (1060bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112100
-----------------------------------------------
mv001	Forward Biobricking of <Cre!
ccataGAATTCatgAGATCTtccaatttactgaccgtacac
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagcaggcg

Construction of {Cre>} Basic Part K112101

PCR mv003/mv004 on pBca9145-Bca1117    (1060bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112101
-----------------------------------------------
mv003	Forward Biobricking of Cre>
ccataGAATTCatgAGATCTatgtccaatttactgaccgtac
mv004	Reverse Biobricking of Cre>
cgttaGGATCCatcgccatcttccagcaggcg

Construction of {<Cre>} Basic Part K112102

PCR mv001/mv004 on pBca9145-Bca1117    (1057bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112102
-----------------------------------------------
mv001	Forward Biobricking of <Cre!
ccataGAATTCatgAGATCTtccaatttactgaccgtacac
mv004	Reverse Biobricking of Cre>
cgttaGGATCCatcgccatcttccagcaggcg

Construction of {r.BamH1!} Basic Part K112103

PCR mv005/mv006 on pBca9145-I716210    (673bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112103
-----------------------------------------------
mv005	Forward Biobricking of r.BamH1!
ccataGAATTCatgAGATCTatggaagtagaaaaagag
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatc

Construction of {r.BamHI>} Basic Part K112104

PCR mv005/mv007 on pBca9145-I716210    (670bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112104
-----------------------------------------------
mv005	Forward Biobricking of r.BamHI!
ccataGAATTCatgAGATCTatggaagtagaaaaagag
mv007	Reverse Biobricking of r.BamHI>
cgttaGGATCCtttgttttcaactttatctttcc

Construction of {<r.BamHI>} Basic Part K112105

PCR mv008/mv007 on pBca9145-I716210    (667bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112105
-----------------------------------------------
mv008	Forward Biobricking of <r.BamHI>
ccataGAATTCatgAGATCTgaagtagaaaaagagtttattac
mv007	Reverse Biobricking of r.BamHI>
cgttaGGATCCtttgttttcaactttatctttcc

Construction of {r.BglII!} Basic Part K112106

PCR mv009/mv010 on pBca9145-Bca1147    (703bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112106
-----------------------------------------------
mv009	Forward Biobricking of r.BglII!
ccataGAATTCatgAGATCTatgaagattgatataacggactataacc
mv010	Reverse Biobricking of r.BglII!
cgttaGGATCCttaatatgtcacgattgttcctc

Construction of {r.BglII>} Basic Part K112107

PCR mv009/mv011 on pBca9145-Bca1147    (700bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112107
-----------------------------------------------
mv009	Forward Biobricking of r.BglII!
ccataGAATTCatgAGATCTatgaagattgatataacggactataacc
mv011	Reverse Biobricking of r.BglII>
cgttaGGATCCatatgtcacgattgttcctc

Construction of {<r.BglII>} Basic Part K112108

PCR mv012/mv011 on pBca9145-Bca1147    (697bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112108
-----------------------------------------------
mv012	Forward Biobricking of <r.BglII>
ccataGAATTCatgAGATCTaagattgatataacggac
mv011	Reverse Biobricking of r.BglII>
cgttaGGATCCatatgtcacgattgttcctc

Construction of {rbs.ligase!} Basic Part K112109

PCR mv013/mv016 on T4 DNA ligase    (269 bp, gp = A)
PCR mv015/mv018 on T4 DNA ligase    (376 bp, gp = B)
PCR mv017/mv020 on T4 DNA ligase    (191 bp, gp = C)
PCR mvo19/mv014 on T4 DNA ligase    (746 bp, gp = D)
---------------------------------------------------
PCR mv013/mv018 on A+B              (622 bp, gp = E)
PCR mv017/mv014 on C+D              (914 bp, gp = F)
---------------------------------------------------
PCR mv013/mv014 on E+F              (1514bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                   (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112109
-----------------------------------------------
mv013	Forward Biobricking of rbs.ligase!
ccataGAATTCatgAGATCTaaactctaaaggatgaac
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg
mv015  Removing the 1st EcoRI site in T4 DNA ligase(F)  gacttcattgaGttcacattagc
mv016  Removing the 1st EcoRI site in T4 DNA ligase(R)  gctaatgtgaaCtcaatgaagtc
mv017  Removing the BglII site in T4 DNA ligase(F)  ctaggattagaCcttcttaagg
mv018  Removing the BglII site in T4 DNA ligase(R)  ccttaagaagGtctaatcctag
mv019  Removing the 2nd EcoRI site in T4 DNA ligase(F)  gctaaagaGttcgccgaagtagc
mv020  Removing the 2nd EcoRI site in T4 DNA ligase(R)  gctacttcggcgaaCtctttagc

Construction of {ligase!} Basic Part K112110

PCR mv015/mv014 on pBca1256-K112109    (1495bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112110
-----------------------------------------------
mv015	Forward Biobricking of ligase!
ccataGAATTCatgAGATCTatgattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {<ligase!} Basic Part K112111

PCR mv016/mv014 on pBca1256-K112109    (1492bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112111
-----------------------------------------------
mv016	Forward Biobricking of <ligase!
ccataGAATTCatgAGATCTattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {ligase>} Basic Part K112112

PCR mv015/mv017 on pBca1256-K112109    (1492bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112112
-----------------------------------------------
mv015	Forward Biobricking of ligase!
ccataGAATTCatgAGATCTatgattcttaaaattctgaacg
mv017	Reverse Biobricking of ligase>
cgttaGGATCCtagaccagttacctcatg

Construction of {<ligase>} Basic Part K112113

PCR mv016/mv017 on pBca1256-K112109    (1489bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112113
-----------------------------------------------
mv016	Forward Biobricking of <ligase!
ccataGAATTCatgAGATCTattcttaaaattctgaacg
mv017	Reverse Biobricking of ligase>
cgttaGGATCCtagaccagttacctcatg