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Revision as of 21:21, 6 June 2008 (view source) Madhvi (Talk | contribs) ← Older edit		Latest revision as of 23:25, 22 September 2008 (view source) Madhvi (Talk | contribs)
(18 intermediate revisions not shown)
Line 1:		Line 1:
		+	[[Template:Team:UC Berkeley/Notebook/MJV construction/corrected assembly vector | patching attR2 site in assembly vector]]<br>
		+	[[Template:Team:UC Berkeley/Notebook/MJV construction/OriV entry vector | OriV entry vector]]
		+
		+
	'''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-Bca1010   (1060bp, gp, EcoRI/BamHI)
	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)		Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
	Product is pBca1256-K112100		Product is pBca1256-K112100
Line 8:		Line 12:
	ccataGAATTCatgAGATCTtccaatttactgaccgtacacc		ccataGAATTCatgAGATCTtccaatttactgaccgtacacc
	mv002 Reverse Biobricking of <Cre!		mv002 Reverse Biobricking of <Cre!
-	cgttaGGATCCttaatcgccatcttccagcaggcg	+	cgttaGGATCCttaatcgccatcttccagc
	</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-Bca1010   (1060bp, EcoRI/BamHI/DpnI)
	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)		Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
	Product is pBca1256-K112101		Product is pBca1256-K112101
Line 26:		Line 30:
	'''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-Bca1010   (1057bp, EcoRI/BamHI/DpnI)
	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)		Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
	Product is pBca1256-K112102		Product is pBca1256-K112102
Line 37:		Line 41:
-	'''Construction of {r.BamH1!} Basic Part K112103'''	+	'''Construction of {r.BamHI!} Basic Part K112103'''
	<pre>		<pre>
	PCR mv005/mv006 on pBca9145-I716210    (673bp, EcoRI/BamHI/DpnI)		PCR mv005/mv006 on pBca9145-I716210    (673bp, EcoRI/BamHI/DpnI)
Line 46:		Line 50:
	ccataGAATTCatgAGATCTatggaagtagaaaaagagtttattactgatgaagc		ccataGAATTCatgAGATCTatggaagtagaaaaagagtttattactgatgaagc
	mv006 Reverse Biobricking of r.BamH1!		mv006 Reverse Biobricking of r.BamH1!
-	cgttaGGATCCctatttgttttcaactttatc	+	cgttaGGATCCctatttgttttcaactttatctttcc
	</pre>		</pre>
Line 68:		Line 72:
	-----------------------------------------------		-----------------------------------------------
	mv008 Forward Biobricking of <r.BamHI>		mv008 Forward Biobricking of <r.BamHI>
-	ccataGAATTCatgAGATCTgaagtagaaaaagagtttattac	+	ccataGAATTCatgAGATCTgaagtagaaaaagagtttattactgatg
	mv007 Reverse Biobricking of r.BamHI>		mv007 Reverse Biobricking of r.BamHI>
	cgttaGGATCCtttgttttcaactttatctttcc		cgttaGGATCCtttgttttcaactttatctttcc
Line 104:		Line 108:
	-----------------------------------------------		-----------------------------------------------
	mv012 Forward Biobricking of <r.BglII>		mv012 Forward Biobricking of <r.BglII>
-	ccataGAATTCatgAGATCTaagattgatataacggac	+	ccataGAATTCatgAGATCTaagattgatataacggactataacc
	mv011 Reverse Biobricking of r.BglII>		mv011 Reverse Biobricking of r.BglII>
	cgttaGGATCCatatgtcacgattgttcctc		cgttaGGATCCatatgtcacgattgttcctc
Line 114:		Line 118:
	PCR mv015/mv018 on T4 DNA ligase    (376 bp, gp = B)		PCR mv015/mv018 on T4 DNA ligase    (376 bp, gp = B)
	PCR mv017/mv020 on T4 DNA ligase    (191 bp, gp = C)		PCR mv017/mv020 on T4 DNA ligase    (191 bp, gp = C)
-	PCR mvo19/mv014 on T4 DNA ligase    (746 bp, gp = D)	+	PCR mv019/mv014 on T4 DNA ligase    (746 bp, gp = D)
	---------------------------------------------------		---------------------------------------------------
	PCR mv013/mv018 on A+B              (622 bp, gp = E)		PCR mv013/mv018 on A+B              (622 bp, gp = E)
Line 198:		Line 202:
	'''Construction of {a~Cre!} Basic Part K112115'''		'''Construction of {a~Cre!} Basic Part K112115'''
	<pre>		<pre>
-	PCR mv025/mv002 on pBca9145-Bca1117   (1067bp, EcoRI/BamHI/DpnI)	+	PCR mv025/mv002 on pBca9145-Bca1010   (1067bp, EcoRI/BamHI/DpnI)
	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)		Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
	Product is pBca1256-K112115		Product is pBca1256-K112115
Line 205:		Line 209:
	ccataGAATTCatgAGATCTttaaatgtccaatttactgaccgtacacc		ccataGAATTCatgAGATCTttaaatgtccaatttactgaccgtacacc
	mv002 Reverse Biobricking of <Cre!		mv002 Reverse Biobricking of <Cre!
-	cgttaGGATCCttaatcgccatcttccagcaggcg	+	cgttaGGATCCttaatcgccatcttccagc
	</pre>		</pre>
-	'''Construction of {a~r.BamH1!} Basic Part K112116'''	+	'''Construction of {a~r.BamHI!} Basic Part K112116'''
	<pre>		<pre>
	PCR mv026/mv006 on pBca9145-I716210    (677bp, EcoRI/BamHI/DpnI)		PCR mv026/mv006 on pBca9145-I716210    (677bp, EcoRI/BamHI/DpnI)
Line 217:		Line 221:
	ccataGAATTCatgAGATCTtagaatggaagtagaaaaagagtttattactgatgaagc		ccataGAATTCatgAGATCTtagaatggaagtagaaaaagagtttattactgatgaagc
	mv006 Reverse Biobricking of r.BamH1!		mv006 Reverse Biobricking of r.BamH1!
-	cgttaGGATCCctatttgttttcaactttatc	+	cgttaGGATCCctatttgttttcaactttatctttcc
	</pre>		</pre>
Line 246:		Line 250:
	'''Construction of {rbs.Cre!} Basic Part K112119'''		'''Construction of {rbs.Cre!} Basic Part K112119'''
	<pre>		<pre>
-	PCR mv030/mv002 on pBca9145-Bca1117   (1082bp, EcoRI/BamHI/DpnI)	+	PCR mv030/mv002 on pBca9145-Bca1010   (1082bp, EcoRI/BamHI/DpnI)
	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)		Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
	Product is pBca1256-K112119		Product is pBca1256-K112119
	-----------------------------------------------		-----------------------------------------------
	mv030 Forward Biobricking of rbs.Cre!		mv030 Forward Biobricking of rbs.Cre!
-	ccataGAATTCatgAGATCTaccattatctgagtgttaaatgtccaatttactgaccgtacacc	+	cgataGAATTCatgAGATCTaccattatctgagtgttaaatgtccaatttactgaccgtacacc
	mv002 Reverse Biobricking of <Cre!		mv002 Reverse Biobricking of <Cre!
-	cgttaGGATCCttaatcgccatcttccagcaggcg	+	cgttaGGATCCttaatcgccatcttccagc
	</pre>		</pre>
-	'''Construction of {rbs.r.BamH1!} Basic Part K112120'''	+	'''Construction of {rbs.r.BamHI!} Basic Part K112120'''
	<pre>		<pre>
	PCR mv031/mv006 on pBca9145-I716210    (692bp, EcoRI/BamHI/DpnI)		PCR mv031/mv006 on pBca9145-I716210    (692bp, EcoRI/BamHI/DpnI)
Line 264:		Line 268:
	-----------------------------------------------		-----------------------------------------------
	mv031 Forward Biobricking of rbs.r.BamH1!		mv031 Forward Biobricking of rbs.r.BamH1!
-	ccataGAATTCatgAGATCTtaagtgggggcaaattagaatggaagtagaaaaagagtttattactgatgaagc	+	cgataGAATTCatgAGATCTtaagtgggggcaaattagaatggaagtagaaaaagagtttattactgatgaagc
	mv006 Reverse Biobricking of r.BamH1!		mv006 Reverse Biobricking of r.BamH1!
-	cgttaGGATCCctatttgttttcaactttatc	+	cgttaGGATCCctatttgttttcaactttatctttcc
	</pre>		</pre>
Line 276:		Line 280:
	-----------------------------------------------		-----------------------------------------------
	mv032 Forward Biobricking of rbs.r.BglII!		mv032 Forward Biobricking of rbs.r.BglII!
-	ccataGAATTCatgAGATCTtggaaggagacactcataatgaagattgatataacggactataaccatgc	+	cgataGAATTCatgAGATCTtggaaggagacactcataatgaagattgatataacggactataaccatgc
	mv010 Reverse Biobricking of r.BglII!		mv010 Reverse Biobricking of r.BglII!
	cgttaGGATCCttaatatgtcacgattgttcctc		cgttaGGATCCttaatatgtcacgattgttcctc
	</pre>		</pre>
		+
		+	'''Construction of {Cre!} Basic Part K112122'''
		+	<pre>
		+	PCR mv003/mv002 on pBca9145-Bca1010    (1063bp, EcoRI/BamHI/DpnI)
		+	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
		+	Product is pBca1256-K112122
		+	-----------------------------------------------
		+	mv003 Forward Biobricking of Cre>
		+	ccataGAATTCatgAGATCTatgtccaatttactgaccgtac
		+	mv002 Reverse Biobricking of <Cre!
		+	cgttaGGATCCttaatcgccatcttccagc
		+	</pre>
		+
		+	'''Construction of {<r.BglII!} Basic Part K112123'''
		+	<pre>
		+	PCR mv012/mv010 on pBca9145-Bca1147    (700bp, EcoRI/BamHI/DpnI)
		+	Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
		+	Product is pBca1256-K112123
		+	-----------------------------------------------
		+	mv012 Forward Biobricking of <r.BglII>
		+	ccataGAATTCatgAGATCTaagattgatataacggactataacc
		+	mv010 Reverse Biobricking of r.BglII!
		+	cgttaGGATCCttaatatgtcacgattgttcctc
		+	</pre>
		+
		+	'''Construction of {rbs.trfA!} Basic Part K112124'''
		+	<pre>
		+	PCR JH102F/JH102R on RP4 genome      (416bp, EcoRI/BamHI/DpnI)
		+	Sub into pBjh1601KC                  (EcoRI/BamHI, 3132+778, L)
		+	Product is pBjh1601KC-K112124
		+	-----------------------------------------------
		+	JH102F Forward Biobricking of {rbs.trfA!}
		+	ccatgGAATTCatgAGATCTgcgaggggcggcttccgctAtgtacaaccag
		+	JH102R Reverse Biobricking of {rbs.trfA!}
		+	cgttaGGATCCtcagagccttccacgcagc
		+	</pre>
		+
		+	'''Construction of {OriV} Basic Part K112125'''
		+	<pre>
		+	PCR mv037/mv038 on pSMART Fosmid Blunt Vector (Lucigen)    (677bp, EcoRI/BamHI/DpnI)
		+	Sub into pBca1256                        (EcoRI/BamHI, 2472+910, L)
		+	Product is pBca1256-K112125
		+	-----------------------------------------------
		+	mv037  Forward biobricking of OriV from pSMART
		+	cgataGAATTCatgAGATCTtgcttcctcgctcactcgac
		+	mv038  Reverse biobricking of OriV form pSMART
		+	gctagGGATCCaagccctcggctggttgc
		+	</pre>
		+
		+	'''Construction of {temp. sensitive promoter} Basic Part K112126'''
		+	<pre>
		+	PCR JH115F/JH115R on pAH57
		+	sub into pBca1256
		+	Product is pBca1256-K112126
		+	-----------------------------------------------
		+	JH115F  Forward biobricking of cI, Pr
		+	cgataGAATTCatgAGATCTgaagattcttgctcaattgttatcagc
		+	JH115R  Reverse biobricking of cI, Pr
		+	gctagGGATCCagaattgatacaacctccttagtacatgc
		+	</pre>
		+
		+	----
		+
		+	<div style="text-align: center;"> [[Team:UC_Berkeley/Notebook/Madhvi_Venkatesh|Madhvi's Notebook]] </div>
		+	<div style="text-align: center;"> [[Team:UC_Berkeley|UC Berkeley Team Homepage]] </div>

---

Latest revision as of 23:25, 22 September 2008

patching attR2 site in assembly vector
OriV entry vector

Construction of {<Cre!} Basic Part K112100

PCR mv001/mv002 on pBca9145-Bca1010    (1060bp, gp, EcoRI/BamHI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112100
-----------------------------------------------
mv001	Forward Biobricking of <Cre!
ccataGAATTCatgAGATCTtccaatttactgaccgtacacc
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

Construction of {Cre>} Basic Part K112101

PCR mv003/mv004 on pBca9145-Bca1010    (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-Bca1010    (1057bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112102
-----------------------------------------------
mv001	Forward Biobricking of <Cre!
ccataGAATTCatgAGATCTtccaatttactgaccgtacacc
mv004	Reverse Biobricking of Cre>
cgttaGGATCCatcgccatcttccagcaggcg

Construction of {r.BamHI!} 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!
ccataGAATTCatgAGATCTatggaagtagaaaaagagtttattactgatgaagc
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatctttcc

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!
ccataGAATTCatgAGATCTatggaagtagaaaaagagtttattactgatgaagc
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>
ccataGAATTCatgAGATCTgaagtagaaaaagagtttattactgatg
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!
ccataGAATTCatgAGATCTatgaagattgatataacggactataaccatgc
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!
ccataGAATTCatgAGATCTatgaagattgatataacggactataaccatgc
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>
ccataGAATTCatgAGATCTaagattgatataacggactataacc
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 mv019/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!
ccataGAATTCatgAGATCTaaactctaaaggatgaacaatg
mv014	Reverse Biobricking of 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

Construction of {ligase!} Basic Part K112110

PCR mv021/mv014 on pBca1256-K112109    (1495bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112110
-----------------------------------------------
mv021	Forward Biobricking of ligase!
ccataGAATTCatgAGATCTatgattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {<ligase!} Basic Part K112111

PCR mv022/mv014 on pBca1256-K112109    (1492bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112111
-----------------------------------------------
mv022	Forward Biobricking of <ligase!
ccataGAATTCatgAGATCTattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {ligase>} Basic Part K112112

PCR mv021/mv023 on pBca1256-K112109    (1492bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112112
-----------------------------------------------
mv021	Forward Biobricking of ligase!
ccataGAATTCatgAGATCTatgattcttaaaattctgaacg
mv023	Reverse Biobricking of ligase>
cgttaGGATCCtagaccagttacctcatg

Construction of {<ligase>} Basic Part K112113

PCR mv022/mv023 on pBca1256-K112109    (1489bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112113
-----------------------------------------------
mv022	Forward Biobricking of <ligase!
ccataGAATTCatgAGATCTattcttaaaattctgaacg
mv023	Reverse Biobricking of ligase>
cgttaGGATCCtagaccagttacctcatg

Construction of {a~ligase!} Basic Part K112114

PCR mv024/mv014 on pBca1256-K112109    (1499bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112114
-----------------------------------------------
mv024	Forward Biobricking of a~ligase!
ccataGAATTCatgAGATCTaacaatgattcttaaaattctgaacg
mv014	Reverse Biobricking of ligase!
cgttaGGATCCtcatagaccagttacctcatg

Construction of {a~Cre!} Basic Part K112115

PCR mv025/mv002 on pBca9145-Bca1010    (1067bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112115
-----------------------------------------------
mv025	Forward Biobricking of a~Cre!
ccataGAATTCatgAGATCTttaaatgtccaatttactgaccgtacacc
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

Construction of {a~r.BamHI!} Basic Part K112116

PCR mv026/mv006 on pBca9145-I716210    (677bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112116
-----------------------------------------------
mv026	Forward Biobricking of a~r.BamH1!
ccataGAATTCatgAGATCTtagaatggaagtagaaaaagagtttattactgatgaagc
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatctttcc

Construction of {a~r.BglII!} Basic Part K112117

PCR mv027/mv010 on pBca9145-Bca1147    (707bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112117
-----------------------------------------------
mv027	Forward Biobricking of a~r.BglII!
ccataGAATTCatgAGATCTcataatgaagattgatataacggactataaccatgc
mv010	Reverse Biobricking of r.BglII!
cgttaGGATCCttaatatgtcacgattgttcctc

Construction of rrnB P1 promoter Basic Part K112118

PCR mv028/mv029 on rrnB native sequence   (721bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                         (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112118
-----------------------------------------------
mv028	Forward Biobricking of rrnB P1 promoter
cgataGAATTCatgAGATCTatgcactaaaacgtatcctacgcccg
mv029   Reverse Biobricking of rrnB P1 promoter
cgttaGGATCCcgccttcccgctacagagtcaagc

Construction of {rbs.Cre!} Basic Part K112119

PCR mv030/mv002 on pBca9145-Bca1010    (1082bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112119
-----------------------------------------------
mv030	Forward Biobricking of rbs.Cre!
cgataGAATTCatgAGATCTaccattatctgagtgttaaatgtccaatttactgaccgtacacc
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

Construction of {rbs.r.BamHI!} Basic Part K112120

PCR mv031/mv006 on pBca9145-I716210    (692bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112120
-----------------------------------------------
mv031	Forward Biobricking of rbs.r.BamH1!
cgataGAATTCatgAGATCTtaagtgggggcaaattagaatggaagtagaaaaagagtttattactgatgaagc
mv006	Reverse Biobricking of r.BamH1!
cgttaGGATCCctatttgttttcaactttatctttcc

Construction of {rbs.r.BglII!} Basic Part K112121

PCR mv032/mv010 on pBca9145-Bca1147    (722bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112121
-----------------------------------------------
mv032	Forward Biobricking of rbs.r.BglII!
cgataGAATTCatgAGATCTtggaaggagacactcataatgaagattgatataacggactataaccatgc
mv010	Reverse Biobricking of r.BglII!
cgttaGGATCCttaatatgtcacgattgttcctc

Construction of {Cre!} Basic Part K112122

PCR mv003/mv002 on pBca9145-Bca1010    (1063bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112122
-----------------------------------------------
mv003	Forward Biobricking of Cre>
ccataGAATTCatgAGATCTatgtccaatttactgaccgtac
mv002	Reverse Biobricking of <Cre!
cgttaGGATCCttaatcgccatcttccagc

Construction of {<r.BglII!} Basic Part K112123

PCR mv012/mv010 on pBca9145-Bca1147    (700bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                      (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112123
-----------------------------------------------
mv012	Forward Biobricking of <r.BglII>
ccataGAATTCatgAGATCTaagattgatataacggactataacc
mv010	Reverse Biobricking of r.BglII!
cgttaGGATCCttaatatgtcacgattgttcctc

Construction of {rbs.trfA!} Basic Part K112124

PCR JH102F/JH102R on RP4 genome      (416bp, EcoRI/BamHI/DpnI)
Sub into pBjh1601KC                  (EcoRI/BamHI, 3132+778, L)
Product is pBjh1601KC-K112124
-----------------------------------------------
JH102F	 Forward Biobricking of {rbs.trfA!}
ccatgGAATTCatgAGATCTgcgaggggcggcttccgctAtgtacaaccag
JH102R	 Reverse Biobricking of {rbs.trfA!}
cgttaGGATCCtcagagccttccacgcagc

Construction of {OriV} Basic Part K112125

PCR mv037/mv038 on pSMART Fosmid Blunt Vector (Lucigen)    (677bp, EcoRI/BamHI/DpnI)
Sub into pBca1256                        (EcoRI/BamHI, 2472+910, L)
Product is pBca1256-K112125
-----------------------------------------------
mv037  Forward biobricking of OriV from pSMART
cgataGAATTCatgAGATCTtgcttcctcgctcactcgac
mv038  Reverse biobricking of OriV form pSMART
gctagGGATCCaagccctcggctggttgc

Construction of {temp. sensitive promoter} Basic Part K112126

PCR JH115F/JH115R on pAH57
sub into pBca1256
Product is pBca1256-K112126
-----------------------------------------------
JH115F  Forward biobricking of cI, Pr
cgataGAATTCatgAGATCTgaagattcttgctcaattgttatcagc
JH115R  Reverse biobricking of cI, Pr
gctagGGATCCagaattgatacaacctccttagtacatgc

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