Template:Team:UC Berkeley/Notebook/SC construction

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 +
<span style="font-family: Century Gothic; font-size: 10pt">
 +
<div style="text-align: right;"> [[Team:UC_Berkeley/Notebook/Sherine_Cheung|Back to Sherine's Notebook]] </div>
 +
<div style="text-align: right;"> [[Team:UC_Berkeley/Notebook| To All Notebooks]] </div>
__TOC__
__TOC__
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== pBca1256-K112600 ==
== pBca1256-K112600 ==
<pre>
<pre>
-
Construction of <AP-tag> basic part K112600
+
Construction of {<AP>} 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, 2472+ 910)
Product is pBca1256-K112600
Product is pBca1256-K112600
----------------------------------------------------
----------------------------------------------------
Line 18: Line 21:
== pBca1256-K112601 ==
== pBca1256-K112601 ==
<pre>
<pre>
-
Construction of <AP-tag! basic part K112601
+
Construction of {<AP!} 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, 2472+ 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 32: Line 35:
== pBca1256-K112602 ==
== pBca1256-K112602 ==
<pre>
<pre>
-
Construction of <FLAG-tag> basic part K112602
+
Construction of {<FLAG>} basic part K112602
-
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
-
-------------------------------------------------------------
+
---------------------------------------------------
SC004    Forward Biobricking of <FLAG-tag>
SC004    Forward Biobricking of <FLAG-tag>
cgataGAATTCatgAGATCTgactacaaggatgacgacg
cgataGAATTCatgAGATCTgactacaaggatgacgacg
Line 46: Line 49:
== pBca1256-K112603 ==
== pBca1256-K112603 ==
<pre>
<pre>
-
Construction of <FLAG-tag! basic part K112603
+
Construction of {<FLAG!} 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, 2472+ 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 64:
<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
-------------------------------
-------------------------------
-
mea035 Fwd biobricking of {rbs_barstar>}  
+
mea035   Fwd biobricking of {rbs_barstar>}  
cgataGAATTCatgAGATCTcataagaaaggagccgcacatg                               
cgataGAATTCatgAGATCTcataagaaaggagccgcacatg                               
-
sc007 Reverse biobricking of {rbs_barstar>}
+
sc007     Reverse biobricking of {rbs_barstar>}                     (Not needed...)
cgttaGGATCCagaaagtatgatggtgatgtcgc   
cgttaGGATCCagaaagtatgatggtgatgtcgc   
</pre>
</pre>
-
== pBca1256-K112403 ==
+
== pBjh1601AC-K112403 ==
<pre>
<pre>
-
Construction of {prepro>} of PhoA basic part K112403         (114 bp, EcoRI/BamHI)
+
Construction 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)                            
-
Product is pBca1256-K112403                                             
+
Sub into pBjh1601AC                      (EcoRI/BamHI, 3195+910 L)
 +
Product is pBjh1601AC-K112403                                             
-------------------------------------------------------------
-------------------------------------------------------------
-
cb10007-phoA-F  Fwd biobricking of {prepro>}
+
cb10007   Fwd biobricking of {prepro>}
-
cgataGAATTCatgAGATCTgtacatggagaaaataaaatgaaacaaagc
+
cgataGAATTCatgAGATCTatgaaacaaagcactattgcactggc
-
cb10008-phoA-R  Reverse biobricking of {prepro>}
+
cb10008   Reverse biobricking of {prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
cgtagGGATCCgggcttttgtcacaggggtaaac
</pre>
</pre>
-
 
+
== pBjh1601AC-K112404  ==
-
== pBca1256-K112404  ==
+
<pre>
<pre>
-
Construction of {prepro>} of LamB basic part K112404             (132 bp, EcoRI/BamHI)
+
Construction 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)                                
-
Product is pBca1256-K112404                                           
+
Sub into pBjh1601AC                    (EcoRI/BamHI, 3195+910, L)                                              
 +
Product is pBjh1601AC-K112404                                           
-------------------------------------------------------------
-------------------------------------------------------------
-
cb10009-lamB-F Fwd biobricking of {prepro>}
+
cb10009   Fwd biobricking of {prepro>}
-
cgataGAATTCatgAGATCTgaaaagcaatgactcaggagatag
+
cgataGAATTCatgAGATCTatgatgattactctgcgcaaac
-
cb10010-lamB-R  Reverse biobricking of {prepro>}
+
cb10010   Reverse biobricking of {prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
cgtagGGATCCcagccattgcctgagcagacattac
</pre>
</pre>
-
 
+
== pBjh1601KC-K112605 ==
-
== pBca1256-K112605 ==
+
<pre>
<pre>
-
Construction of {a~prepro>} of PhoA basic part K112605             (100 bp, EcoRI/BamHI)
+
Construction of {a~PhoA>} basic part K112605  
-
PCR sc008/cb10008-phoA-R                                           (EcoRI/BamHI, 2472+910, L)
+
       
-
Sub into pBca1256
+
PCR sc008/cb10008-phoA-R on MG1655            (100 bp, EcoRI/BamHI)                             
-
Product is pBca1256-K112605
+
Sub into pBjh1601KC                            (EcoRI/BamHI, 3132+910, L)
 +
Product is pBjh1601KC-K112605
------------------------------------------------------
------------------------------------------------------
-
sc008 Fwd biobricking of {a~prepro>}
+
sc008     Fwd biobricking of {a~prepro>}
-
cgataGAATTCatgAGATCT taaa Atgaaacaaagcactattgcac
+
cgataGAATTCatgAGATCTtaaaGtgaaacaaagcactattgcactggc
-
cb10008-phoA-R Reverse biobricking of {a~prepro>}
+
cb10008   Reverse biobricking of {a~prepro>}
-
cgtagGGATCCgggcttttgtcacaggggtaaac
+
cgtagGGATCCgggcttttgtcacaggggtaaac                                
</pre>
</pre>
-
== pBca1256-K112606 ==
+
== pBjh1601CK-K112606 ==
<pre>
<pre>
-
Construction of {a~prepro>} of LamB basic part K112606                (111 bp, EcoRI/BamHI)
+
Construction of {a~LamB>} basic part K112606  
-
PCR 009/cb10010-lamB-R                                                (EcoRI/BamHI, 2472+910, L)
+
                  
-
Sub into pBca1256
+
PCR sc009/cb10010-lamB-R on MG1655            (111 bp, EcoRI/BamHI)                              
-
Product is pBca1256- K112606
+
Sub into pBjh1601CK                            (EcoRI/BamHI, 3134+910, L)
 +
Product is pBjh1601CK-K112606
--------------------------------------------------
--------------------------------------------------
-
sc009 Fwd biobricking of {a~prepro>}
+
sc009     Fwd biobricking of {a~prepro>}
-
cgataGAATTCatgAGATCT taga atgatgattactctgc
+
cgataGAATTCatgAGATCTtagaatgatgattactctgcgc
-
cb10010-lamB-R Reverse biobricking of {a~prepro>}
+
cb10010   Reverse biobricking of {a~prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
cgtagGGATCCcagccattgcctgagcagacattac
</pre>
</pre>
-
== pBca1256-K112607  ==
+
== pBjh1601KC-K112607  ==
<pre>
<pre>
-
Construction of {rbs_prepro>} of PhoA basic part K112607              (113 bp, EcoRI/BamHI)
+
Construction of {rbs_PhoA>} basic part K112607               
-
PCR sc010/cb10008-PhoA-R                                               (EcoRI/BamHI, 2472+910, L)
+
 
-
Sub into pBca1256
+
PCR sc010/cb10008-PhoA-R on MG1655            (113 bp, EcoRI/BamHI)                       
-
Product is pBca1256-K112607
+
Sub into pBjh1601KC                            (EcoRI/BamHI, 3132+910, L)
 +
Product is pBjh1601KC-K112607
------------------------------------------------------
------------------------------------------------------
-
sc010 Fwd biobricking of {rbs_prepro>}
+
sc010     Fwd biobricking of {rbs_prepro>}
-
cgataGAATTCatgAGATCT gtacatggagaaaataaaAtgaaacaaag
+
cgataGAATTCatgAGATCTgtacatggagaaaataaaAtgaaacaaagcac
-
cb10008-phoA-R  Reverse biobricking of {rbs_prepro>}
+
cb10008   Reverse biobricking of {rbs_prepro>}
cgtagGGATCCgggcttttgtcacaggggtaaac
cgtagGGATCCgggcttttgtcacaggggtaaac
</pre>
</pre>
-
== pBca1256-K112608 ==
+
== pBjh1601KC-K112608 ==
<pre>
<pre>
-
Construction of {rbs_prepro>} of LamB basic part K112608         (126 bp, EcoRI/BamHI)
+
Construction of {rbs_LamB>} basic part K112608  
-
PCR sc011/cb10010-lamB-R                                         (EcoRI/BamHI, 2472+910, L)
+
     
-
Sub into pBca1256
+
PCR sc011/cb10010-lamB-R on E.coli K12 MG1655            (126 bp, EcoRI/BamHI)           
-
Product is pBca1256-K112608
+
Sub into pBjh1601KC                                        (EcoRI/BamHI, 3132+910, L)
 +
Product is pBjh1601KC-K112608
------------------------------------------------------
------------------------------------------------------
-
sc011 Fwd biobricking of {rbs_prepro>}
+
sc011     Fwd biobricking of {rbs_prepro>}
-
cgataGAATTCatgAGATCT caatgactcaggagataga atgatgattac
+
cgataGAATTCatgAGATCTcaatgactcaggagatagaatg
-
cb10010-lamB-R Reverse biobricking of {rbs_prepro>}
+
cb10010   Reverse biobricking of {rbs_prepro>}
cgtagGGATCCcagccattgcctgagcagacattac
cgtagGGATCCcagccattgcctgagcagacattac
</pre>
</pre>
 +
== pBjh1601CA-K112609 ==
 +
<pre>
 +
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                               
 +
</pre>
 +
---------------------------------------------------------------------------------------------------------------
 +
 +
 +
== pBca1256-K112610 ==
 +
<pre>
 +
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
 +
</pre>
 +
 +
== pBca1256-K112611 ==
 +
<pre>
 +
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>}
 +
ccagtGGATCCagccattgcctgagcagac                               
 +
</pre>
 +
 +
== pBca1256-K112612 ==
 +
<pre>
 +
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
 +
</pre>
 +
 +
== pBca1256-K112613 ==
 +
<pre>
 +
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>}
 +
ccagtGGATCCagccattgcctgagcagac                               
 +
</pre>
 +
 +
== pBca1256-K112614 ==
 +
<pre>
 +
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                             
 +
</pre>
 +
 +
== pBca1256-K112615 ==
 +
<pre>
 +
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>}
 +
ccagtGGATCCagccattgcctgagcagac                             
 +
</pre>
 +
 +
== pBca1102-Bca9194 Assembly part ==
 +
<pre>
 +
PCR ca998/mea037 on Bca9194                          (1610 bp, EcoRI/BamHI?)
 +
Sub into {<tag!}.{b1006} composite part (CK vector)  (EcoRI/BamHI,____+__)
 +
Product is _______________
 +
---------------------------------------------------------
 +
ca998    Fwd biobricking
 +
gtatcacgaggcagaatttcag
 +
mea037    Reverse biobricking of {<phoA>}
 +
cgttaGGATCCCTTCAGGCCCAGCGCCGCTTTCATG
 +
</pre>
----
----
<div style="text-align: center;"> [[Team:UC_Berkeley/Notebook/Sherine_Cheung|Sherine Cheung]] </div>
<div style="text-align: center;"> [[Team:UC_Berkeley/Notebook/Sherine_Cheung|Sherine Cheung]] </div>
<div style="text-align: center;"> [[Team:UC_Berkeley|Back to Berkeley Team Homepage]] </div>
<div style="text-align: center;"> [[Team:UC_Berkeley|Back to Berkeley Team Homepage]] </div>
 +
<div style="text-align: center;"> [[Team:UC_Berkeley/Notebook| All Notebooks]] </div>

Latest revision as of 20:23, 8 August 2008

Back to Sherine's Notebook
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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>}
ccagtGGATCCagccattgcctgagcagac                                

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>}
ccagtGGATCCagccattgcctgagcagac                                

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>}
ccagtGGATCCagccattgcctgagcagac                              

pBca1102-Bca9194 Assembly part

PCR ca998/mea037 on Bca9194                           (1610 bp, EcoRI/BamHI?)
Sub into {<tag!}.{b1006} composite part (CK vector)   (EcoRI/BamHI,____+__) 
Product is _______________
---------------------------------------------------------
ca998     Fwd biobricking 
gtatcacgaggcagaatttcag
mea037    Reverse biobricking of {<phoA>}
cgttaGGATCCCTTCAGGCCCAGCGCCGCTTTCATG

Sherine Cheung
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