Template:Team:UC Berkeley/Notebook/MJV construction

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Revision as of 02:21, 10 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!
ccataGAATTCatgAGATCTtccaatttactgaccgtacacc
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

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!
ccataGAATTCatgAGATCTtccaatttactgaccgtacacc
mv004	Reverse Biobricking of Cre>
cgttaGGATCCatcgccatcttccagcaggcg

Construction of {r.BamHI!} 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!
ccataGAATTCatgAGATCTatggaagtagaaaaagagtttattactgatgaagc
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatctttcc

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!
ccataGAATTCatgAGATCTatggaagtagaaaaagagtttattactgatgaagc
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!
ccataGAATTCatgAGATCTatgaagattgatataacggactataaccatgc
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!
ccataGAATTCatgAGATCTatgaagattgatataacggactataaccatgc
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 mv019/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!
ccataGAATTCatgAGATCTaaactctaaaggatgaacaatg
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 mv021/mv014 on pBca1256-K112109    (1495bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112110
-----------------------------------------------
mv021	Forward Biobricking of ligase!
ccataGAATTCatgAGATCTatgattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {<ligase!} Basic Part K112111

PCR mv022/mv014 on pBca1256-K112109    (1492bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112111
-----------------------------------------------
mv022	Forward Biobricking of <ligase!
ccataGAATTCatgAGATCTattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {ligase>} Basic Part K112112

PCR mv021/mv023 on pBca1256-K112109    (1492bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112112
-----------------------------------------------
mv021	Forward Biobricking of ligase!
ccataGAATTCatgAGATCTatgattcttaaaattctgaacg
mv023	Reverse Biobricking of ligase>
cgttaGGATCCtagaccagttacctcatg

Construction of {<ligase>} Basic Part K112113

PCR mv022/mv023 on pBca1256-K112109    (1489bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112113
-----------------------------------------------
mv022	Forward Biobricking of <ligase!
ccataGAATTCatgAGATCTattcttaaaattctgaacg
mv023	Reverse Biobricking of ligase>
cgttaGGATCCtagaccagttacctcatg

Construction of {a~ligase!} Basic Part K112114

PCR mv024/mv014 on pBca1256-K112109    (1499bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112114
-----------------------------------------------
mv024	Forward Biobricking of a~ligase!
ccataGAATTCatgAGATCTaacaatgattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {a~Cre!} Basic Part K112115

PCR mv025/mv002 on pBca9145-Bca1117    (1067bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112115
-----------------------------------------------
mv025	Forward Biobricking of a~Cre!
ccataGAATTCatgAGATCTttaaatgtccaatttactgaccgtacacc
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

Construction of {a~r.BamHI!} Basic Part K112116

PCR mv026/mv006 on pBca9145-I716210    (677bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112116
-----------------------------------------------
mv026	Forward Biobricking of a~r.BamH1!
ccataGAATTCatgAGATCTtagaatggaagtagaaaaagagtttattactgatgaagc
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatctttcc

Construction of {a~r.BglII!} Basic Part K112117

PCR mv027/mv010 on pBca9145-Bca1147    (707bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112117
-----------------------------------------------
mv027	Forward Biobricking of a~r.BglII!
ccataGAATTCatgAGATCTcataatgaagattgatataacggactataaccatgc
mv010	Reverse Biobricking of r.BglII!
cgttaGGATCCttaatatgtcacgattgttcctc

Construction of rrnB P1 promoter Basic Part K112118

PCR mv028/mv029 on rrnB native sequence   (721bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                         (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112118
-----------------------------------------------
mv028	Forward Biobricking of rrnB P1 promoter
cgataGAATTCatgAGATCTatgcactaaaacgtatcctacgcccg
mv029   Reverse Biobricking of rrnB P1 promoter
cgttaGGATCCcgccttcccgctacagagtcaagc

Construction of {rbs.Cre!} Basic Part K112119

PCR mv030/mv002 on pBca9145-Bca1117    (1082bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112119
-----------------------------------------------
mv030	Forward Biobricking of rbs.Cre!
cgataGAATTCatgAGATCTaccattatctgagtgttaaatgtccaatttactgaccgtacacc
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

Construction of {rbs.r.BamHI!} Basic Part K112120

PCR mv031/mv006 on pBca9145-I716210    (692bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112120
-----------------------------------------------
mv031	Forward Biobricking of rbs.r.BamH1!
cgataGAATTCatgAGATCTtaagtgggggcaaattagaatggaagtagaaaaagagtttattactgatgaagc
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatc

Construction of {rbs.r.BglII!} Basic Part K112121

PCR mv032/mv010 on pBca9145-Bca1147    (722bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112121
-----------------------------------------------
mv032	Forward Biobricking of rbs.r.BglII!
cgataGAATTCatgAGATCTtggaaggagacactcataatgaagattgatataacggactataaccatgc
mv010	Reverse Biobricking of r.BglII!
cgttaGGATCCttaatatgtcacgattgttcctc