Template:Team:UC Berkeley/Notebook/SC construction

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(Difference between revisions)
Line 7: Line 7:
Wobble PCR SC001/SC002            (66 bp, EcoRI/BamHI)
Wobble PCR SC001/SC002            (66 bp, EcoRI/BamHI)
-
Sub into pBca1256                (EcoRI/BamHI, 2227+ 910)
+
Sub into pBca1256                (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112600
Product is pBca1256-K112600
----------------------------------------------------
----------------------------------------------------
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Wobble PCR SC001/SC003            (69 bp, EcoRI/BamHI)
Wobble PCR SC001/SC003            (69 bp, EcoRI/BamHI)
-
Sub into pBca1256                (EcoRI/BamHI, 2227+ 910)
+
Sub into pBca1256                (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112601
Product is pBca1256-K112601
--------------------------------------------------
--------------------------------------------------
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Wobble PCR SC004/SC005            (45 bp, EcoRI/BamHI)
Wobble PCR SC004/SC005            (45 bp, EcoRI/BamHI)
-
Sub into pBca1256                (EcoRI/BamHI, 2227+ 910)
+
Sub into pBca1256                (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112602
Product is pBca1256-K112602
---------------------------------------------------
---------------------------------------------------
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Wobble PCR SC004/SC006            (48 bp, EcoRI/BamHI)
Wobble PCR SC004/SC006            (48 bp, EcoRI/BamHI)
-
Sub into pBca1256                (EcoRI/BamHI, 2227+ 910)
+
Sub into pBca1256                (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112603
Product is pBca1256-K112603
-----------------------------------------------------
-----------------------------------------------------
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CR cb10007/cb10008 on MG1655            (95 bp, EcoRI/BamHI)                               
CR cb10007/cb10008 on MG1655            (95 bp, EcoRI/BamHI)                               
-
Sub into pBjh1601AC                      (EcoRI/BamHI, 2472+910, L)
+
Sub into pBjh1601AC                      (EcoRI/BamHI, 3195+910 L)
Product is pBjh1601AC-K112403                                             
Product is pBjh1601AC-K112403                                             
-------------------------------------------------------------
-------------------------------------------------------------
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PCR cb10009/cb10010 on MG1655            (107 bp, EcoRI/BamHI)                                 
PCR cb10009/cb10010 on MG1655            (107 bp, EcoRI/BamHI)                                 
-
Sub into pBjh1601AC                       (EcoRI/BamHI, 2472+910, L)                                               
+
Sub into pBjh1601AC                     (EcoRI/BamHI, 3195+910, L)                                               
Product is pBjh1601AC-K112404                                           
Product is pBjh1601AC-K112404                                           
-------------------------------------------------------------
-------------------------------------------------------------
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PCR sc008/cb10008-phoA-R on MG1655            (100 bp, EcoRI/BamHI)                               
PCR sc008/cb10008-phoA-R on MG1655            (100 bp, EcoRI/BamHI)                               
-
Sub into pBjh1601KC                            (EcoRI/BamHI, 2472+910, L)
+
Sub into pBjh1601KC                            (EcoRI/BamHI, 3132+910, L)
Product is pBjh1601KC-K112605
Product is pBjh1601KC-K112605
------------------------------------------------------
------------------------------------------------------
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PCR sc009/cb10010-lamB-R on MG1655            (111 bp, EcoRI/BamHI)                               
PCR sc009/cb10010-lamB-R on MG1655            (111 bp, EcoRI/BamHI)                               
-
Sub into pBjh1601CK                            (EcoRI/BamHI, 2472+910, L)
+
Sub into pBjh1601CK                            (EcoRI/BamHI, 3134+910, L)
Product is pBjh1601CK-K112606
Product is pBjh1601CK-K112606
--------------------------------------------------
--------------------------------------------------
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PCR sc010/cb10008-PhoA-R on MG1655            (113 bp, EcoRI/BamHI)                         
PCR sc010/cb10008-PhoA-R on MG1655            (113 bp, EcoRI/BamHI)                         
-
Sub into pBjh1601KC                            (EcoRI/BamHI, 2472+910, L)
+
Sub into pBjh1601KC                            (EcoRI/BamHI, 3132+910, L)
Product is pBjh1601KC-K112607
Product is pBjh1601KC-K112607
------------------------------------------------------
------------------------------------------------------
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PCR sc011/cb10010-lamB-R on E.coli K12 MG1655            (126 bp, EcoRI/BamHI)             
PCR sc011/cb10010-lamB-R on E.coli K12 MG1655            (126 bp, EcoRI/BamHI)             
-
Sub into pBjh1601KC                                        (EcoRI/BamHI, 2472+910, L)
+
Sub into pBjh1601KC                                        (EcoRI/BamHI, 3132+910, L)
Product is pBjh1601KC-K112608
Product is pBjh1601KC-K112608
------------------------------------------------------
------------------------------------------------------

Revision as of 20:42, 28 July 2008

Contents


pBca1256-K112600 

Construction of {<AP>} basic part K112600

Wobble PCR SC001/SC002            (66 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112600
----------------------------------------------------
SC001F    Forward Biobricking of <AP-tag>
cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag
SC002R    Reverse Biobricking of <AP-tag>
ccagtGGATCCttcatgccattcaattttctgcgcttcaaaaatatcg

pBca1256-K112601 

Construction of {<AP!} basic part K112601

Wobble PCR SC001/SC003            (69 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112601
--------------------------------------------------
SC001    Forward Biobricking of <AP-tag!
cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag
SC003    Reverse Biobricking of <AP-tag!
ccagtGGATCCttattcatgccattcaattttctgcgcttcaaaaatatcg

pBca1256-K112602 

Construction of {<FLAG>} basic part K112602

Wobble PCR SC004/SC005            (45 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112602
---------------------------------------------------
SC004     Forward Biobricking of <FLAG-tag>
cgataGAATTCatgAGATCTgactacaaggatgacgacg
SC005     Reverse Biobricking of <FLAG-tag>
ccagtGGATCCcttgtcgtcgtcatccttgtagtc

pBca1256-K112603 

Construction of {<FLAG!} basic part K112603

Wobble PCR SC004/SC006            (48 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2472+ 910)
Product is pBca1256-K112603
-----------------------------------------------------
SC004    Forward Biobricking of <FLAG-tag!
cgataGAATTCatgAGATCTgactacaaggatgacgacg
SC006    Reverse Biobricking of <FLAG-tag!
ccagtGGATCCttacttgtcgtcgtcatccttgtagtc

pBca1256-K112604 

Construction of {rbs_barstar>} basic part K112604

PCR mea035/sc007 on Bacillus amyloliquefaciens gen.            (320 bp, EcoRI/BamHI)
Sub into pBca1256                                              (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112604
-------------------------------
mea035    Fwd biobricking of {rbs_barstar>} 
cgataGAATTCatgAGATCTcataagaaaggagccgcacatg                              
sc007     Reverse biobricking of {rbs_barstar>}                     (Not needed...)
cgttaGGATCCagaaagtatgatggtgatgtcgc

pBjh1601AC-K112403 

Construction of {PhoA>} basic part K112403
          
CR cb10007/cb10008 on MG1655            (95 bp, EcoRI/BamHI)                              
Sub into pBjh1601AC                       (EcoRI/BamHI, 3195+910 L)
Product is pBjh1601AC-K112403                                            
-------------------------------------------------------------
cb10007    Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgaaacaaagcactattgcactggc
cb10008    Reverse biobricking of {prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac

pBjh1601AC-K112404 

Construction of {LamB>} basic part K112404   
           
PCR cb10009/cb10010 on MG1655            (107 bp, EcoRI/BamHI)                                 
Sub into pBjh1601AC                     (EcoRI/BamHI, 3195+910, L)                                               
Product is pBjh1601AC-K112404                                           
-------------------------------------------------------------
cb10009    Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgatgattactctgcgcaaac
cb10010    Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBjh1601KC-K112605 

Construction of {a~PhoA>} basic part K112605    
         
PCR sc008/cb10008-phoA-R on MG1655            (100 bp, EcoRI/BamHI)                              
Sub into pBjh1601KC                             (EcoRI/BamHI, 3132+910, L)
Product is pBjh1601KC-K112605
------------------------------------------------------
sc008      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtaaaGtgaaacaaagcactattgcactggc
cb10008    Reverse biobricking of {a~prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac

pBjh1601CK-K112606 

Construction of {a~LamB>} basic part K112606 
                
PCR sc009/cb10010-lamB-R on MG1655            (111 bp, EcoRI/BamHI)                               
Sub into pBjh1601CK                             (EcoRI/BamHI, 3134+910, L)
Product is pBjh1601CK-K112606
--------------------------------------------------
sc009      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtagaatgatgattactctgcgc
cb10010    Reverse biobricking of {a~prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBjh1601KC-K112607 

Construction of {rbs_PhoA>} basic part K112607               

PCR sc010/cb10008-PhoA-R on MG1655            (113 bp, EcoRI/BamHI)                         
Sub into pBjh1601KC                             (EcoRI/BamHI, 3132+910, L)
Product is pBjh1601KC-K112607
------------------------------------------------------
sc010      Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaagcac
cb10008    Reverse biobricking of {rbs_prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac

pBjh1601KC-K112608 

Construction of {rbs_LamB>} basic part K112608   
      
PCR sc011/cb10010-lamB-R on E.coli K12 MG1655            (126 bp, EcoRI/BamHI)             
Sub into pBjh1601KC                                        (EcoRI/BamHI, 3132+910, L)
Product is pBjh1601KC-K112608
------------------------------------------------------
sc011      Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
cb10010    Reverse biobricking of {rbs_prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBjh1601CA-K112609 

Construction of {a~PhoA>} basic part K112609 (K112605 replacement)
         
PCR sc012/cb10008-phoA-R on MG1655            (100 bp, EcoRI/BamHI)                              
Sub into pBjh1601CA                             (EcoRI/BamHI, 2472+910, L)
Product is pBjh1601CA-K112605
------------------------------------------------------
sc012      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtaaaAtgaaacaaagcactattgcactggc
cb10008    Reverse biobricking of {a~prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac


pBca1256-K112610 

Construction of {PhoA>} basic part K112610
         
PCR cb10007/sc013 on MG1655                   (94 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112610
------------------------------------------------------
cb10007    Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgaaacaaagcactattgcactggc
sc013    Reverse biobricking of {prepro>}
ccagtGGATCCggcttttgtcacaggggtaaac

pBca1256-K112611 

Construction of {LamB>} basic part K112611 
         
PCR cb10009/sc014 on MG1655                   (107 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
------------------------------------------------------
cb10009    Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgatgattactctgcgcaaac
sc014    Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBca1256-K112612 

Construction of {a~PhoA>} basic part K112612 
         
PCR sc012/sc013 on MG1655                     (98 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
------------------------------------------------------
sc012      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtaaaAtgaaacaaagcactattgcactggc
sc013    Reverse biobricking of {prepro>}
ccagtGGATCCggcttttgtcacaggggtaaac

pBca1256-K112613 

Construction of {a~LamB>} basic part K112613
         
PCR sc009/sc014 on MG1655                     (111 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
------------------------------------------------------
sc009      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtagaatgatgattactctgcgc
sc014    Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBca1256-K112614 

Construction of {rbs_PhoA>} basic part K112614
         
PCR sc010/sc013 on MG1655                     (112 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
------------------------------------------------------
sc010      Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaagcac
sc013    Reverse biobricking of {prepro>}
ccagtGGATCCggcttttgtcacaggggtaaac

pBca1256-K112615 

Construction of {rbs_LamB>} basic part K112615

PCR sc011/sc014 on MG1655                     (126 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
------------------------------------------------------
sc011      Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
sc014    Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac


Sherine Cheung
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