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Template:Team:UC Berkeley/Notebook/MJV construction
From 2008.igem.org
(Difference between revisions)
| Line 1: | Line 1: | ||
| - | '''Construction of <Cre! Basic Part K112100''' | + | '''Construction of {<Cre!} Basic Part K112100''' |
<pre> | <pre> | ||
PCR mv001/mv002 on pBca9145-Bca1117 (1060bp, EcoRI/BamHI/DpnI) | PCR mv001/mv002 on pBca9145-Bca1117 (1060bp, EcoRI/BamHI/DpnI) | ||
| Line 6: | Line 6: | ||
----------------------------------------------- | ----------------------------------------------- | ||
mv001 Forward Biobricking of <Cre! | mv001 Forward Biobricking of <Cre! | ||
| - | + | ccataGAATTCatgAGATCTtccaatttactgaccgtacac | |
mv002 Reverse Biobricking of <Cre! | mv002 Reverse Biobricking of <Cre! | ||
cgttaGGATCCttaatcgccatcttccagcaggcg | cgttaGGATCCttaatcgccatcttccagcaggcg | ||
</pre> | </pre> | ||
| - | '''Construction of Cre> Basic Part K112101''' | + | '''Construction of {Cre>} Basic Part K112101''' |
<pre> | <pre> | ||
PCR mv003/mv004 on pBca9145-Bca1117 (1060bp, EcoRI/BamHI/DpnI) | PCR mv003/mv004 on pBca9145-Bca1117 (1060bp, EcoRI/BamHI/DpnI) | ||
| Line 18: | Line 18: | ||
----------------------------------------------- | ----------------------------------------------- | ||
mv003 Forward Biobricking of Cre> | mv003 Forward Biobricking of Cre> | ||
| - | + | ccataGAATTCatgAGATCTatgtccaatttactgaccgtac | |
mv004 Reverse Biobricking of Cre> | mv004 Reverse Biobricking of Cre> | ||
cgttaGGATCCatcgccatcttccagcaggcg | cgttaGGATCCatcgccatcttccagcaggcg | ||
| Line 24: | Line 24: | ||
| - | '''Construction of <Cre> Basic Part K112102''' | + | '''Construction of {<Cre>} Basic Part K112102''' |
<pre> | <pre> | ||
PCR mv001/mv004 on pBca9145-Bca1117 (1057bp, EcoRI/BamHI/DpnI) | PCR mv001/mv004 on pBca9145-Bca1117 (1057bp, EcoRI/BamHI/DpnI) | ||
| Line 31: | Line 31: | ||
----------------------------------------------- | ----------------------------------------------- | ||
mv001 Forward Biobricking of <Cre! | mv001 Forward Biobricking of <Cre! | ||
| - | + | ccataGAATTCatgAGATCTtccaatttactgaccgtacac | |
mv004 Reverse Biobricking of Cre> | mv004 Reverse Biobricking of Cre> | ||
cgttaGGATCCatcgccatcttccagcaggcg | cgttaGGATCCatcgccatcttccagcaggcg | ||
| + | </pre> | ||
| + | |||
| + | |||
| + | '''Construction of {r.BamH1!} Basic Part K112103''' | ||
| + | <pre> | ||
| + | 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 | ||
| + | </pre> | ||
| + | |||
| + | '''Construction of {r.BamHI>} Basic Part K112104''' | ||
| + | <pre> | ||
| + | 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 | ||
| + | </pre> | ||
| + | |||
| + | '''Construction of {<r.BamHI>} Basic Part K112105''' | ||
| + | <pre> | ||
| + | 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 | ||
| + | </pre> | ||
| + | |||
| + | '''Construction of {r.BglII!} Basic Part K112106''' | ||
| + | <pre> | ||
| + | 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 | ||
| + | </pre> | ||
| + | |||
| + | '''Construction of {r.BglII>} Basic Part K112107''' | ||
| + | <pre> | ||
| + | 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 | ||
| + | </pre> | ||
| + | |||
| + | '''Construction of {<r.BglII>} Basic Part K112108''' | ||
| + | <pre> | ||
| + | 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 | ||
| + | </pre> | ||
| + | |||
| + | '''Construction of {rbs.ligase!} Basic Part K112109''' | ||
| + | <pre> | ||
| + | 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 rbs.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 | ||
</pre> | </pre> | ||
Revision as of 01:42, 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 rbs.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
