Template:Team:UC Berkeley/Notebook/SC construction

From 2008.igem.org

(Difference between revisions)
Line 6: Line 6:
Construction of <AP-tag> basic part K112600
Construction of <AP-tag> basic part K112600
-
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, 2227+ 910)
Product is pBca1256-K112600
Product is pBca1256-K112600
----------------------------------------------------
----------------------------------------------------
Line 20: Line 20:
Construction of <AP-tag! basic part K112601
Construction of <AP-tag! basic part K112601
-
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, 2227+ 910)
Product is pBca1256-K112601
Product is pBca1256-K112601
--------------------------------------------------
--------------------------------------------------
-
SC001     Forward Biobricking of <AP-tag!
+
SC001   Forward Biobricking of <AP-tag!
cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag
cgataGAATTCatgAGATCTggcctgaacgatatttttgaagcgcag
-
SC003     Reverse Biobricking of <AP-tag!
+
SC003   Reverse Biobricking of <AP-tag!
ccagtGGATCCttattcatgccattcaattttctgcgcttcaaaaatatcg
ccagtGGATCCttattcatgccattcaattttctgcgcttcaaaaatatcg
</pre>
</pre>
Line 48: Line 48:
Construction of <FLAG-tag! basic part K112603
Construction of <FLAG-tag! basic part K112603
-
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, 2227+ 910)
Product is pBca1256-K112603
Product is pBca1256-K112603
-----------------------------------------------------
-----------------------------------------------------
-
SC004   Forward Biobricking of <FLAG-tag!
+
SC004   Forward Biobricking of <FLAG-tag!
cgataGAATTCatgAGATCTgactacaaggatgacgacg
cgataGAATTCatgAGATCTgactacaaggatgacgacg
-
SC006   Reverse Biobricking of <FLAG-tag!
+
SC006   Reverse Biobricking of <FLAG-tag!
ccagtGGATCCttacttgtcgtcgtcatccttgtagtc
ccagtGGATCCttacttgtcgtcgtcatccttgtagtc
</pre>
</pre>
Line 61: Line 61:
<pre>
<pre>
Construction of {rbs_barstar>} basic part 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)
+
PCR mea035/sc007 on Bacillus amyloliquefaciens gen.           (320 bp, EcoRI/BamHI)
 +
Sub into pBca1256                                             (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112604
Product is pBca1256-K112604
-------------------------------
-------------------------------
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== pBca1256-K112403 ==
== pBca1256-K112403 ==
<pre>
<pre>
-
Construction of {prepro>} of PhoA basic part K112403         (95 bp, EcoRI/BamHI)
+
Construction of {prepro>} of PhoA basic part K112403
-
CR cb10007/cb10008 on MG1655                                  (EcoRI/BamHI, 2472+910, L)
+
         
-
Sub into pBca1256
+
CR cb10007/cb10008 on MG1655            (95 bp, EcoRI/BamHI)                            
 +
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112403                                             
Product is pBca1256-K112403                                             
-------------------------------------------------------------
-------------------------------------------------------------
-
cb10007-phoA-F  Fwd biobricking of {prepro>}
+
cb10007   Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgaaacaaagcactattgcactggc
cgataGAATTCatgAGATCTatgaaacaaagcactattgcactggc
-
cb10008-phoA-R  Reverse biobricking of {prepro>}
+
cb10008   Reverse biobricking of {prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
cgtagGGATCCgggcttttgtcacaggggtaaac
</pre>
</pre>
Line 86: Line 88:
== pBca1256-K112404  ==
== pBca1256-K112404  ==
<pre>
<pre>
-
Construction of {prepro>} of LamB basic part K112404             (107 bp, EcoRI/BamHI)
+
Construction of {prepro>} of LamB basic part K112404  
-
PCR cb10009/cb10010 on MG1655                                    (EcoRI/BamHI, 2472+910, L)
+
         
-
Sub into pBca1256                                                             
+
PCR cb10009/cb10010 on MG1655            (107 bp, EcoRI/BamHI)                                
 +
Sub into pBca1256                        (EcoRI/BamHI, 2472+910, L)                                              
Product is pBca1256-K112404                                           
Product is pBca1256-K112404                                           
-------------------------------------------------------------
-------------------------------------------------------------
-
cb10009-lamB-F  Fwd biobricking of {prepro>}
+
cb10009   Fwd biobricking of {prepro>}
cgataGAATTCatgAGATCTatgatgattactctgcgcaaac
cgataGAATTCatgAGATCTatgatgattactctgcgcaaac
-
cb10010-lamB-R  Reverse biobricking of {prepro>}
+
cb10010   Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
cgtagGGATCCcagccattgcctgagcagacattac
</pre>
</pre>
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== pBca1256-K112605 (needs to be redone -- replace with Atg)==
== pBca1256-K112605 (needs to be redone -- replace with Atg)==
<pre>
<pre>
-
Construction of {a~prepro>} of PhoA basic part K112605             (100 bp, EcoRI/BamHI)
+
Construction of {a~prepro>} of PhoA basic part K112605  
-
PCR sc008/cb10008-phoA-R on MG1655                                 (EcoRI/BamHI, 2472+910, L)
+
       
-
Sub into pBca1256
+
PCR sc008/cb10008-phoA-R on MG1655           (100 bp, EcoRI/BamHI)                             
 +
Sub into pBca1256                            (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112605
Product is pBca1256-K112605
------------------------------------------------------
------------------------------------------------------
-
sc008   Fwd biobricking of {a~prepro>}
+
sc008     Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtaaaGtgaaacaaagcactattgcactggc
cgataGAATTCatgAGATCTtaaaGtgaaacaaagcactattgcactggc
-
cb10008-phoA-R   Reverse biobricking of {a~prepro>}
+
cb10008    Reverse biobricking of {a~prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac                                 
cgtagGGATCCgggcttttgtcacaggggtaaac                                 
</pre>
</pre>
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== pBca1256-K112606 ==
== pBca1256-K112606 ==
<pre>
<pre>
-
Construction of {a~prepro>} of LamB basic part K112606                (111 bp, EcoRI/BamHI)
+
Construction of {a~prepro>} of LamB basic part K112606  
-
PCR sc009/cb10010-lamB-R on MG1655                                     (EcoRI/BamHI, 2472+910, L)
+
                  
-
Sub into pBca1256
+
PCR sc009/cb10010-lamB-R on MG1655           (111 bp, EcoRI/BamHI)                             
 +
Sub into pBca1256                            (EcoRI/BamHI, 2472+910, L)
Product is pBca1256- K112606
Product is pBca1256- K112606
--------------------------------------------------
--------------------------------------------------
-
sc009   Fwd biobricking of {a~prepro>}
+
sc009     Fwd biobricking of {a~prepro>}
cgataGAATTCatgAGATCTtagaatgatgattactctgcgc
cgataGAATTCatgAGATCTtagaatgatgattactctgcgc
-
cb10010-lamB-R   Reverse biobricking of {a~prepro>}
+
cb10010    Reverse biobricking of {a~prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
cgtagGGATCCcagccattgcctgagcagacattac
</pre>
</pre>
Line 125: Line 130:
== pBca1256-K112607  ==
== pBca1256-K112607  ==
<pre>
<pre>
-
Construction of {rbs_prepro>} of PhoA basic part K112607              (113 bp, EcoRI/BamHI)
+
Construction of {rbs_prepro>} of PhoA basic part K112607               
-
PCR sc010/cb10008-PhoA-R on MG1655                                     (EcoRI/BamHI, 2472+910, L)
+
 
-
Sub into pBca1256
+
PCR sc010/cb10008-PhoA-R on MG1655           (113 bp, EcoRI/BamHI)                       
 +
Sub into pBca1256                            (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112607
Product is pBca1256-K112607
------------------------------------------------------
------------------------------------------------------
-
sc010   Fwd biobricking of {rbs_prepro>}
+
sc010     Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaagcac
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaagcac
-
cb10008-phoA-R  Reverse biobricking of {rbs_prepro>}
+
cb10008   Reverse biobricking of {rbs_prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
cgtagGGATCCgggcttttgtcacaggggtaaac
</pre>
</pre>
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== pBca1256-K112608 ==
== pBca1256-K112608 ==
<pre>
<pre>
-
Construction of {rbs_prepro>} of LamB basic part  K112608         (126 bp, EcoRI/BamHI)
+
Construction of {rbs_prepro>} of LamB basic part  K112608  
-
PCR sc011/cb10010-lamB-R on E.coli K12 MG1655                     (EcoRI/BamHI, 2472+910, L)
+
     
-
Sub into pBjh1601AK
+
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
Product is pBca1256-K112608
------------------------------------------------------
------------------------------------------------------
-
sc011   Fwd biobricking of {rbs_prepro>}
+
sc011     Fwd biobricking of {rbs_prepro>}
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
-
cb10010-lamB-R  Reverse biobricking of {rbs_prepro>}
+
cb10010   Reverse biobricking of {rbs_prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
cgtagGGATCCcagccattgcctgagcagacattac
</pre>
</pre>

Revision as of 21:24, 9 July 2008

Contents


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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