Template:Team:UC Berkeley/Notebook/SC construction

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pBca1256-K112600

Construction of <AP-tag> basic part K112600

Wobble PCR SC001/SC002            (66 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2227+ 910)
Product is pBca1256-K112600
----------------------------------------------------
SC001F    Forward Biobricking of <AP-tag>
cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag
SC002R    Reverse Biobricking of <AP-tag>
ccagtGGATCCttcatgccattcaattttctgcgcttcaaaaatatcg

pBca1256-K112601

Construction of <AP-tag! basic part K112601

Wobble PCR SC001/SC003            (69 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2227+ 910)
Product is pBca1256-K112601
--------------------------------------------------
SC001    Forward Biobricking of <AP-tag!
cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag
SC003    Reverse Biobricking of <AP-tag!
ccagtGGATCCttattcatgccattcaattttctgcgcttcaaaaatatcg

pBca1256-K112602

Construction of <FLAG-tag> basic part K112602

Wobble PCR SC004/SC005            (45 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2227+ 910)
Product is pBca1256-K112602
---------------------------------------------------
SC004     Forward Biobricking of <FLAG-tag>
cgataGAATTCatgAGATCTgactacaaggatgacgacg
SC005     Reverse Biobricking of <FLAG-tag>
ccagtGGATCCcttgtcgtcgtcatccttgtagtc

pBca1256-K112603

Construction of <FLAG-tag! basic part K112603

Wobble PCR SC004/SC006            (48 bp, EcoRI/BamHI)
Sub into pBca1256                 (EcoRI/BamHI, 2227+ 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  

pBca1256-K112403

Construction of {prepro>} of PhoA basic part K112403
          
CR cb10007/cb10008 on MG1655            (95 bp, EcoRI/BamHI)                              
Sub into pBca1256                       (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112403                                            
-------------------------------------------------------------
cb10007    Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgaaacaaagcactattgcactggc
cb10008    Reverse biobricking of {prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac

pBca1256-K112404

Construction of {prepro>} of LamB basic part K112404   
           
PCR cb10009/cb10010 on MG1655            (107 bp, EcoRI/BamHI)                                 
Sub into pBca1256                        (EcoRI/BamHI, 2472+910, L)                                               
Product is pBca1256-K112404                                           
-------------------------------------------------------------
cb10009    Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgatgattactctgcgcaaac
cb10010    Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBca1256-K112605 (needs to be redone -- replace with Atg)

Construction of {a~prepro>} of PhoA basic part K112605    
         
PCR sc008/cb10008-phoA-R on MG1655            (100 bp, EcoRI/BamHI)                              
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
------------------------------------------------------
sc008      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtaaaGtgaaacaaagcactattgcactggc
cb10008    Reverse biobricking of {a~prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac                                 

pBca1256-K112606

Construction of {a~prepro>} of LamB basic part K112606 
                
PCR sc009/cb10010-lamB-R on MG1655            (111 bp, EcoRI/BamHI)                               
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256- K112606
--------------------------------------------------
sc009      Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtagaatgatgattactctgcgc
cb10010    Reverse biobricking of {a~prepro>}
cgtagGGATCCcagccattgcctgagcagacattac

pBca1256-K112607

Construction of {rbs_prepro>} of PhoA basic part K112607               

PCR sc010/cb10008-PhoA-R on MG1655            (113 bp, EcoRI/BamHI)                         
Sub into pBca1256                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112607
------------------------------------------------------
sc010      Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaagcac
cb10008    Reverse biobricking of {rbs_prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac

pBca1256-K112608

Construction of {rbs_prepro>} of LamB basic part K112608   
      
PCR sc011/cb10010-lamB-R on E.coli K12 MG1655            (126 bp, EcoRI/BamHI)             
Sub into pBca1256                                        (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112608
------------------------------------------------------
sc011      Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
cb10010    Reverse biobricking of {rbs_prepro>}
cgtagGGATCCcagccattgcctgagcagacattac



Sherine Cheung
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