Paris/Freezer

From 2007.igem.org

< Paris(Difference between revisions)
(→Digestion Products)
(→PCR products)
 
(49 intermediate revisions not shown)
Line 39: Line 39:
| style="background: #ccffcc;" |P3
| style="background: #ccffcc;" |P3
|DGAT-1
|DGAT-1
 +
|27 ForDGAT1
 +
|13 DPst1-DGAT-R
 +
|Miniprep pKS::DGAT
 +
|1087
|
|
|
|
|
|
-
|
+
|[[Paris/July_31|July 31]]
-
|
+
-
|
+
-
|
+
-
|
+
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |P4
| style="background: #ccffcc;" |P4
|DGAT-2
|DGAT-2
 +
|12 DPst1-DGAT
 +
|27 Rev-DGAT1
 +
|Miniprep pKS::DGAT
 +
|396
|
|
|
|
|
|
-
|
+
|[[Paris/July_31|July 31]]
-
|
+
-
|
+
-
|
+
-
|
+
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |P5
| style="background: #ccffcc;" |P5
Line 135: Line 135:
|
|
|[[Paris/July 27| July 27]],[[Paris/July 30| July 30]]
|[[Paris/July 27| July 27]],[[Paris/July 30| July 30]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |P13
 +
|DGAT deltaPst-1
 +
| ForDGAT1
 +
| Megaprimer DGAT2
 +
| DNA plasmid (pKS::DGAT):
 +
|1443
 +
|
 +
|
 +
|
 +
|[[Paris/August_1| August 1]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |P14
 +
|DGAT deltaPst-1 biobrick
 +
| ForDGAT1bb
 +
| RevDGAT1bb
 +
| DGAT deltaPst1
 +
|1504
 +
|
 +
|
 +
|
 +
|[[Paris/August_2| August 2]][[Paris/October 18| Oct. 18]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |P15
 +
| PacI [pTet>>lox-GFP-loxmRFP] XmaI
 +
| O47
 +
| O48
 +
| [pTet>>lox-GFP-loxmRFP] L59.6
 +
|2kb
 +
|
 +
|
 +
|
 +
|[[Paris/August_2| August 2]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |P16
 +
| PrefixBB-frt1-CmR-frt2-SufixBB
 +
| O51
 +
| O52
 +
| pY-2C-IntC
 +
|1kb
 +
|
 +
|
 +
|
 +
|[[Paris/October 15| Oct. 15]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |P17
 +
| PrefixBB-frt1-CmR-frt2-T1T2-SufixBB
 +
| O51
 +
| O53
 +
| pY-2C-IntC
 +
|>1kb
 +
|
 +
|
 +
|
 +
|[[Paris/October 18| Oct. 18]]
|}
|}
-
 
http://partsregistry.org/images/partbypart/icon_terminator.png
http://partsregistry.org/images/partbypart/icon_terminator.png
http://partsregistry.org/images/partbypart/icon_regulatory.png
http://partsregistry.org/images/partbypart/icon_regulatory.png
Line 161: Line 215:
|Strong constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23100 SpecifiedComponents</partinfo>
|Strong constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23100 SpecifiedComponents</partinfo>
|
|
-
| [[Paris/July 15| July 15]]
+
| [[Paris/July 15| July 15]], [[Paris/August 17| Aug. 17]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |MP2
| style="background: #ccffcc;" |MP2
Line 167: Line 221:
|Medium constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23107 SpecifiedComponents</partinfo>
|Medium constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23107 SpecifiedComponents</partinfo>
|MP2.2  
|MP2.2  
-
| [[Paris/July 15| July 15]]
+
| [[Paris/July 15| July 15]], [[Paris/August 17| Aug. 17]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |MP3
| style="background: #ccffcc;" |MP3
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart>
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart>
|Double terminator (clones 1 + 2) <partinfo>B0015 SpecifiedComponents</partinfo>
|Double terminator (clones 1 + 2) <partinfo>B0015 SpecifiedComponents</partinfo>
-
|MP3.1
+
|MP3.3, MP3.4
-
| [[Paris/July 16| July 16]]
+
| [[Paris/August 14| August 14]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |MP4
| style="background: #ccffcc;" |MP4
Line 228: Line 282:
|
|
| [[Paris/July 27| July 27]]
| [[Paris/July 27| July 27]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |MP13
 +
|<bbpart>BBa_J61002</bbpart>
 +
|ptet-mRFP <partinfo>J61002 SpecifiedComponents</partinfo>
 +
|MP13.1, MP13.2
 +
| [[Paris/August 14| August 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |MP14
 +
|<bbpart>BBa_J61025</bbpart>
 +
|Medium constitutive promoter in J61002 (clones 1 + 2) <partinfo>BBa_J61025 SpecifiedComponents</partinfo>
 +
|MP14.1, MP14.2
 +
| [[Paris/August 14| August 14]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |L1
| style="background: #ccffcc;" |L1
|<bbpart>BBa_I0500</bbpart> in <bbpart>J61002</bbpart>
|<bbpart>BBa_I0500</bbpart> in <bbpart>J61002</bbpart>
|construct with pBad promotor regulating expression of mRFP <partinfo>I0500 SpecifiedComponents</partinfo>
|construct with pBad promotor regulating expression of mRFP <partinfo>I0500 SpecifiedComponents</partinfo>
-
| L1".1, L1".2
+
|L1".1, L1".2
 +
| [[Paris/August 14| August 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |
 +
|AttB
 +
|
 +
|MP15
 +
| [[Paris/August 14| August 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |
 +
|AttP
 +
|Finished
 +
|MP16
| [[Paris/July 25| July 25]], [[Paris/July 27| July 27]], [[Paris/August 7| Aug 7]]
| [[Paris/July 25| July 25]], [[Paris/July 27| July 27]], [[Paris/August 7| Aug 7]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 255: Line 333:
|
|
|
|
-
|
+
|[[Paris/July 25| July 25]], [[Paris/July 30| July 30]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |L10''
| style="background: #ccffcc;" |L10''
Line 261: Line 339:
|
|
|
|
-
|
+
|[[Paris/July 25| July 25]], [[Paris/July 30| July 30]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |L12
| style="background: #ccffcc;" |L12
Line 271: Line 349:
|[[Paris/July 30| July 30]]
|[[Paris/July 30| July 30]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
-
| style="background: #ccffcc;" |L25
+
| style="background: #ccffcc;" |L26
-
|lox71-KmR-lox66 in <bbpart>pSB1A2</bbpart>
+
|araC/pbad>>RBS-Cre in <bbpart>J61002</bbpart>
-
|ligation of lox71-KmR-lox66 as a biobrick in pSB1A2 plasmid
+
|ligation ([[Paris/August 13| Aug.13]]) of RBS-Cre as a CDS behind pbad in <bbpart>J61002</bbpart>
-
http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png
+
|L26.1 (no more) & L26.2
-
http://partsregistry.org/wiki/images/2/2e/Part_icon_generator.png
+
|[[Paris/August 17| Aug. 17]]
-
http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png
+
|- style="background: #cccccc;"
-
|L25.1
+
| style="background: #ccffcc;" |L29
-
|[[Paris/July 30| July 30]]
+
|gfptripart-lox66 <bbpart>pSB1A2</bbpart>
 +
|ligation ([[Paris/August 13| Aug.13]]) of gfp-tripart in front of lox66
 +
|L29.1 & L29.2
 +
|[[Paris/August 17| Aug. 17]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L30
 +
|dapAp>>mRFP in <bbpart>J61002</bbpart>
 +
|dapAp promotor placed([[Paris/August 13| Aug.13]]) upstream of mRFP <bbpart>J61002</bbpart>
 +
|L30A1 & L30A2
 +
|[[Paris/August 17| Aug. 17]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L31
 +
|lox71-<bbpart>B0015</bbpart>(T) in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L31.1 & L31.3
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L32
 +
|<bbpart>B0015</bbpart>(T)-lox66 in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L32.1 & L32.2
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L34
 +
|pTet>>lox71-gfptripart in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L34.1 & L34.2
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L35
 +
|araC/pBad>>lox71-gfptripart in <bbpart>pSB1A2</bbpart>
 +
|il manque la seq. de 740-1410
 +
|L35.1 & L35.2
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L36
 +
|lox66-mRFP
 +
|
 +
|L36.1 & L36.2
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L37
 +
|lox66-RBS-dapAcoli
 +
|
 +
|L37.1 & L37.2
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L39
 +
|RBS-dapAcoli cloned in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L39.1 & L39.2
 +
|[[Paris/August 20| Aug. 20]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L46
 +
|pTet>>lox71-B0015(T) in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L46.1
 +
|[[Paris/September 18| Sep. 18]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L53
 +
|B0015(T)-lox66-RBSDapAcoli in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L53.7
 +
|[[Paris/September 18| Sep. 18]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L56
 +
|B0015(T)-lox66-RBSdapAsubtilis in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L56.3 (attention, seq a partiellement foiré, refaire seq)
 +
|[[Paris/September 18| Sep. 18]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L59
 +
|pTet>>lox71-gfptripart-lox66-mRFP <bbpart>pSB1A2</bbpart>
 +
|
 +
|L59.6 (attention, seq avec O40 a foiré, mais les extrémités sont bonnes, je pense que c ok car les colonies sont vertes)
 +
|[[Paris/September 18| Sep. 18]]
 +
|- style="background: #cccccc;"
 +
| style="background: #ccffcc;" |L64
 +
|RBSdapAsubtilis cloned in <bbpart>pSB1A2</bbpart>
 +
|
 +
|L564.1 & L64.2
 +
|[[Paris/September 18| Sep. 18]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |MPT1
| style="background: #ccffcc;" |MPT1
-
|
+
|Lox66-DapAColi
|pTOPO P5
|pTOPO P5
|MPT1.1, MPT1.2
|MPT1.1, MPT1.2
-
|Lox66-DapAColi
+
|[[Paris/August 7| Aug 7]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
| style="background: #ccffcc;" |MPT3
| style="background: #ccffcc;" |MPT3
Line 303: Line 462:
|MPT5.1, .2 &.3
|MPT5.1, .2 &.3
|[[Paris/August 7| Aug 7]]
|[[Paris/August 7| Aug 7]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |MP15
 +
|pKD46
 +
|
 +
|
 +
|[[Paris/October 2| Oct 2]]
|}
|}
Line 331: Line 496:
|2096
|2096
|Vector with the J23100 promoter ready for insertion of a RBS/CDS insert  
|Vector with the J23100 promoter ready for insertion of a RBS/CDS insert  
-
|
+
|Ø (24/09/07)
|34
|34
|[[Paris/July 17| July 17]]
|[[Paris/July 17| July 17]]
Line 353: Line 518:
|2096
|2096
|Vector with the J23107 promoter ready for insertion of a RBS/CDS insert  
|Vector with the J23107 promoter ready for insertion of a RBS/CDS insert  
-
|
+
|Ø (24/09/07)
|32
|32
|[[Paris/July 17| July 17]]
|[[Paris/July 17| July 17]]
Line 562: Line 727:
|Ø D22.1
|Ø D22.1
|Open pSB1A2 vector for BioBrick EcoRI/PstI insertio
|Open pSB1A2 vector for BioBrick EcoRI/PstI insertio
-
|D22.1 Ø
+
|Ø (24/09/07)
|D22.2:59
|D22.2:59
|[[Paris/July 23| July 23]],[[Paris/August 13 | Aug. 13 ]]
|[[Paris/July 23| July 23]],[[Paris/August 13 | Aug. 13 ]]
Line 764: Line 929:
|[[Paris/August 6 | Aug. 6 ]]
|[[Paris/August 6 | Aug. 6 ]]
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
-
| style="background: #ccffcc;" |D41
+
| style="background: #FF99FF;" |D41
|araC/pBad-J61002 plasmid SP dig.
|araC/pBad-J61002 plasmid SP dig.
|L1.2, L1".1 & L1".2 miniprep product  
|L1.2, L1".1 & L1".2 miniprep product  
Line 796: Line 961:
|
|
|[[Paris/August 13| August 13]]
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D44
 +
|<bbpart>B0015</bbpart> terminator BI
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> (MP3.3 Aug.14)
 +
|XbaI
 +
|Pst1
 +
|
 +
|terminator B0015 extracted for use as a BI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D45
 +
|<bbpart>B0015</bbpart> terminator FI
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> (MP3.4 Aug.14)
 +
|EcoRI
 +
|SpeI
 +
|
 +
|terminator B0015 extracted for use as a FI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D46
 +
|<bbpart>B0015</bbpart> BV
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> (MP3.3 Aug.14)
 +
|SpeI
 +
|Pst1
 +
|
 +
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> plasmid extracted for use as a BV
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D47
 +
|<bbpart>pSB1A2</bbpart> XP dig.
 +
|<bbpart>pSB1A2</bbpart> carrying Cre ORF (MP9.1)
 +
|XbaI
 +
|PstI
 +
|
 +
|<bbpart>pSB1A2</bbpart> open vector ready for cloning of BI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D48
 +
|<bbpart>pSB1A2</bbpart> ES dig.
 +
|<bbpart>pSB1A2</bbpart> carrying Cre ORF (MP9.2)
 +
|EcoRI
 +
|SpeI
 +
|
 +
|<bbpart>pSB1A2</bbpart> open vector ready for cloning of FI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D49
 +
|lox71-gfptripart BI
 +
|lox71-gfptripart in <bbpart>pSB1A2</bbpart> (L12.2)
 +
|XbaI
 +
|Pst1
 +
|
 +
|lox71-gfptripart extracted for use as a BI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D50
 +
|<bbpart>BBa_J61002</bbpart> pTet SP dig.
 +
|<bbpart>BBa_J61002</bbpart> (MP13.1 [[Paris/August 14| Aug. 14]])
 +
|SpeI
 +
|PstI
 +
|
 +
|<bbpart>BBa_J61002</bbpart> pTet promoter vector ready for insertion of CDS as a BI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D51
 +
|RBS-dapAcoli BBinsert
 +
|pTOPO of RBS-dapAcoli (MPT4.3)
 +
|EcoRI
 +
|Pst1
 +
|
 +
|RBS-dapAcoli extracted for cloning as a biobrick
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D52
 +
|RBS-dapAcoli BI
 +
|pTOPO of RBS-dapAcoli (MPT4.3)
 +
|XbaI
 +
|Pst1
 +
|
 +
|RBS-dapAcoli extracted for use as a BI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D53
 +
|RBS-dapAsubtilis BBinsert
 +
|pTOPO of RBS-dapAsubtilis (MPT5.1)
 +
|EcoRI
 +
|Pst1
 +
|
 +
|RBS-dapAsubtilis extracted for cloning as a biobrick
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D54
 +
|RBS-dapAsubtilis BI
 +
|pTOPO of RBS-dapAsubtilis (MPT5.1)
 +
|XbaI
 +
|Pst1
 +
|
 +
|RBS-dapAsubtilis extracted for use as a BI
 +
|Ø (24/09/07)
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D55
 +
|RBS-dapAsubtilis FI
 +
|pTOPO of RBS-dapAsubtilis (MPT5.2)
 +
|EcoRI
 +
|SpeI
 +
|
 +
|RBS-dapAsubtilis extracted for use as a FI
 +
|
 +
|
 +
|[[Paris/August 16| August 16]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D57
 +
|lox71-B0015(T) BI
 +
|(L31.1, L31.3) lox71-B0015(T) <bbpart>pSB1A2</bbpart>
 +
|XbaI
 +
|Pst1
 +
|
 +
|lox71-B0015(T) extracted for use as a BI <bbpart>pSB1A2</bbpart>
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D58
 +
|B0015(T)-lox66 BV
 +
|(L32.1, L32.2) B0015(T)-lox66 BV
 +
|SpeI
 +
|PstI
 +
|
 +
|B0015(T)-lox66 for use as a BV <bbpart>pSB1A2</bbpart>
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D59
 +
|pTet>>lox71-gfptripart BI
 +
|(L34.1, L34.2) pTet>>lox71-gfptripart <bbpart>BBa_J61002</bbpart>
 +
|XbaI
 +
|PstI
 +
|
 +
|pTet>>lox71-gfptripart for use as a BI
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;"  |D61
 +
|lox66-mRFP FV
 +
|(L36.1, L36.2) lox66-mRFP
 +
|EcoRI
 +
|XbaI
 +
|
 +
|lox66-mRFP part (strongRBS-ORF-T) ready for use as a forward vector
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D62
 +
|lox66-RBSDapAcoli BI
 +
|(L37.1, L37.2) lox66-RBSDapAcoli
 +
|XbaI
 +
|PstI
 +
|
 +
|lox66-RBSDapAcoli ready for use as a Backward insert
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D63
 +
|RBSDapAcoli BI
 +
|(L39.1, L39.2) RBSDapAcoli
 +
|XbaI
 +
|PstI
 +
|
 +
|RBSDapAcoli BI (for insertion behind T-lox66)
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D64
 +
|FRT-CmR-FRT <bbpart>BBa_J61025</bbpart> BV
 +
|(MP14.1) <bbpart>BBa_J61025</bbpart>
 +
|SpeI
 +
|PstI
 +
|
 +
|FRT-CmR-FRT <bbpart>BBa_J61025</bbpart> (for backward introduction of a casette for Wanner K insertion)
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D65
 +
|FRT-CmR-FRT <bbpart>BBa_J61025</bbpart> FV
 +
|(MP14.1) <bbpart>BBa_J61025</bbpart>
 +
|EcoR
 +
|XbaI
 +
|
 +
|FRT-CmR-FRT <bbpart>BBa_J61025</bbpart> (for forward introduction of a casette for Wanner K insertion)
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D68
 +
|attB BV
 +
|(MP15.1)
 +
|SpeI
 +
|PstI
 +
|
 +
|vector containing attB for use as a BV
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D69
 +
|gfptripart-lox66 BV
 +
|(L29.1) gfptripart-lox66 <bbpart>pSB1A2</bbpart>
 +
|SpeI
 +
|PstI
 +
|
 +
|
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D70
 +
|pTet>>lox71-gfptripart FI
 +
|(L34.1, L34.2) pTet>>lox71-gfptripart <bbpart>BBa_J61002</bbpart>
 +
|EcoRI
 +
|SpeI
 +
|
 +
|pTet>>lox71-gfptripart for use as a FI
 +
|
 +
|
 +
|[[Paris/September 14| Sep. 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D71
 +
|araC/pBad>>lox71-gfptripart FI
 +
|L35.1 araC/pBad>>lox71-gfptripart <bbpart>BBa_J61002</bbpart>
 +
|EcoRI
 +
|SpeI
 +
|
 +
|araC/pBad>>lox71-gfptripart for use as a FI
 +
|Ø (24/09/07)
 +
|
 +
|[[Paris/September 14| Sep. 14]][[Paris/September 17| Sep. 17]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D72
 +
|<bbpart>pSB2K3</bbpart> BBvector
 +
|MP16 <bbpart>pSB2K3</bbpart>::attP
 +
|EcoRI
 +
|PstI
 +
|
 +
|Low copy number plasmid, BB vector
 +
|
 +
|
 +
|[[Paris/September 17| Sep. 17]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D73
 +
|<bbpart>pSB2K3</bbpart> ES open vector
 +
|MP16 <bbpart>pSB2K3</bbpart>::attP
 +
|EcoRI
 +
|SpeI
 +
|
 +
|Comparison of digestion efficiency between Pst and Spe in NEB2
 +
|
 +
|
 +
|[[Paris/September 17| Sep. 17]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D76
 +
|DGAT BB
 +
|P14 DGAT
 +
|EcoRI
 +
|PstI
 +
|
 +
|DGAT
 +
|Ø (24/09/07)
 +
|
 +
|[[Paris/September 17| Sep. 17]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |D77
 +
|araC/pBad>>lox71-gfptripart BI
 +
|D60 (.1 & .2) digestion product
 +
|FspI
 +
|AhdI
 +
|
 +
|in D60, the extracted fragment & the open plasmid have the same size, it is thus necessary to remove the plasmid by digestion
 +
|
 +
|
 +
|[[Paris/September 17| Sep. 17]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;"  |D78
 +
|araC/pBad>>lox71-gfptripart FI
 +
|D71 (.1 & .2) digestion product
 +
|FspI
 +
|AhdI
 +
|
 +
|in D71, the extracted fragment & the open plasmid have the same size, it is thus necessary to remove the plasmid by digestion
 +
|
 +
|
 +
|[[Paris/September 17| Sep. 17]]
|}
|}
Line 834: Line 1,318:
|
|
|[[Paris/July 23| July 23]]
|[[Paris/July 23| July 23]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #FF99FF;" |D66
 +
|(MP12.1, MP12.2) <bbpart>BBa_P1003</bbpart> in <bbpart>pSB1A2</bbpart>
 +
|EcoR
 +
|XbaI
 +
|
 +
|KanR <bbpart>BBa_J61025</bbpart> FV (for forward introduction of a casette for Wanner K insertion). The strange thing is that this digestion produces 2 visible products: ~1000 bp & ~2500 bp!!!
 +
|[[Paris/September 17| Sep.17]], [[Paris/September 27| Sep.27]]
|}
|}
Line 976: Line 1,468:
|<bbpart>pSB1A2</bbpart>::Lox71-kmR-lox66
|<bbpart>pSB1A2</bbpart>::Lox71-kmR-lox66
|Lox71-kmR-lox66 (L25)  
|Lox71-kmR-lox66 (L25)  
-
|
+
|sequence not good (thrown away)
|?
|?
|- style="background: #cccccc;"   
|- style="background: #cccccc;"   
Line 1,014: Line 1,506:
|
|
|[[Paris/August 14| August 14]]
|[[Paris/August 14| August 14]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S29
 +
|L26
 +
|Insertion of RBS-Cre behind araC/pBad
 +
|
 +
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S30
 +
|L27
 +
|Insertion of ftsZ behind araC/pBad
 +
|
 +
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S31
 +
|L28
 +
|Insertion of ftsAZ behind araC/pBad
 +
|
 +
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S32
 +
|L29
 +
|Insertion of gfp-tripart in front of lox66
 +
|
 +
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S33A
 +
|L30A
 +
|cloning dapAp promotor in J61002
 +
|
 +
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S33B
 +
|L30B
 +
|cloning dapAp promotor in J61002
 +
|
 +
|[[Paris/August 13| August 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S34
 +
|L31
 +
|Insertion of B0015 terminator behind lox71
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S35
 +
|L32
 +
|Insertion of B0015 terminator in front of lox66
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S36
 +
|L34
 +
|pTet>>lox71-gfptripart
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S37
 +
|L35
 +
|araC/pBad>>lox71-gfptripart
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S38
 +
|L36
 +
|lox66-mRFP
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S39
 +
|L37
 +
|lox66-RBS-dapAcoli BI
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S40
 +
|L39
 +
|RBS-dapAcoli cloned in pSB1A2
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S41
 +
|L40
 +
|RBS-dapAsubtilis cloned in pSB1A2
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S42
 +
|L41
 +
|lox71-ftsZ cloned in pSB1A2
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S43
 +
|L42
 +
|lox71-ftsA-Z cloned in pSB1A2
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S44
 +
|L43
 +
|RBS-dapAsubtilis cloned in pSB1A2
 +
|
 +
|[[Paris/August 22| August 22]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S45A
 +
|L30A
 +
|DapAp>>mRFP within W121 strain
 +
|
 +
|[[Paris/August 25| August 25]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S45B
 +
|L30B
 +
|DapAp>>mRFP within W121 strain
 +
|
 +
|[[Paris/August 25| August 25]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S48
 +
|pKD13
 +
|FRT-KanR-FRT; Wanner plasmid
 +
|
 +
|[[Paris/September 18| Sep. 18]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S49
 +
|pKD46
 +
|Red recombination system; Wanner plasmid
 +
|
 +
|[[Paris/September_13| Sep. 13]]
 +
|- style="background: #cccccc;" 
 +
| style="background: #ccffcc;" |S50
 +
|L30 plasmid in FR781
 +
|dapAp>>mRFP plasmid
 +
|
 +
|[[Paris/September_28| Sep. 28]]
|}
|}

Latest revision as of 16:18, 26 November 2007

Contents

PCR products

PCR products
Number Name Oligo F Oligo R Matrix Length Comments Product Left ? []ng/µL Date
P1 Lox71-FtsA-FtsZ-1 O3 Lox71-FtsA-F O4 DEcoR1-FtsZ-R MG1655 1537 Ø
P2 FtsZ-2 O5 DEcoR1-FtsZ-F O2 FtsZ-R MG1655 1088 Ø
P3 DGAT-1 27 ForDGAT1 13 DPst1-DGAT-R Miniprep pKS::DGAT 1087 July 31
P4 DGAT-2 12 DPst1-DGAT 27 Rev-DGAT1 Miniprep pKS::DGAT 396 July 31
P5 Lox66-DapAColi O6 Lox66-DapAColi-F O7 DapAColi-R MG1655 985 Ø July 17, July 30
P6 Lox66-DapASubtilis O8 Lox66-DapASubt-F O9 DapASubt-R B.Subtilis 168 979 July 17, July 27, July 30
P7 Lox71-FtsZ O1 Lox71-FtsZ-F O2 FtsZ-R MG1655 1260 EcoRI site in FtsZ not mutated July 17, July 30
P8 RBS-DapAColi O20 RBS-DapAColi-F O7 DapAColi-R MG1655 952 B0030-DapAColi July 23, July 30
P9 RBS-DapASubtilis O21 RBS-DapASubt-F O9 DapASubt-R B.Subtilis 168 946 B0030-DapASubtilis July 23, July 27
P10 Lox71-FtsA-FtsZ O3 Lox71-FtsA-F O2 FtsZ-R MG1655 2580 EcoRI site in FtsZ not mutated July 23
P11 Lox71-KmR-Lox66 O10 Lox71-KmR-F O11 Lox66-KmR-R MP12 1084 July 27, July 30
P13 DGAT deltaPst-1 ForDGAT1 Megaprimer DGAT2 DNA plasmid (pKS::DGAT): 1443 August 1
P14 DGAT deltaPst-1 biobrick ForDGAT1bb RevDGAT1bb DGAT deltaPst1 1504 August 2 Oct. 18
P15 PacI [pTet>>lox-GFP-loxmRFP] XmaI O47 O48 [pTet>>lox-GFP-loxmRFP] L59.6 2kb August 2
P16 PrefixBB-frt1-CmR-frt2-SufixBB O51 O52 pY-2C-IntC 1kb Oct. 15
P17 PrefixBB-frt1-CmR-frt2-T1T2-SufixBB O51 O53 pY-2C-IntC >1kb Oct. 18

http://partsregistry.org/images/partbypart/icon_terminator.png http://partsregistry.org/images/partbypart/icon_regulatory.png http://partsregistry.org/images/partbypart/icon_rbs.png http://partsregistry.org/images/partbypart/icon_reporter.png http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png http://partsregistry.org/wiki/images/2/2e/Part_icon_generator.png http://partsregistry.org/wiki/images/a/ab/Part_icon_plasmid.png

Plasmids: MiniPrep Products

Plasmids: MiniPrep Products
Number Name Description Products in use Date
MP1 pJ23100 in J61002 Strong constitutive promoter in J61002 (clones 1 + 2) July 15, Aug. 17
MP2 pJ23107 in J61002 Medium constitutive promoter in J61002 (clones 1 + 2) MP2.2 July 15, Aug. 17
MP3 B0015 in pSB1AK3 Double terminator (clones 1 + 2) MP3.3, MP3.4 August 14
MP4 I0500 in pSB2K3 AraC pBAD (clones 1 + 2) July 16
MP5 BBa_B0030 in pSB1A2 Strong RBS (clones 1 + 2) July 18
MP6 BBa_E0422 in pSB1A2 ECFP (RBS+LVA+Term) (clones 1 + 2) MP6'.1, MP6'.2 July 18, July 24
MP7 BBa_E0241 in pSB1A2 PoPs to GFP converter (clones 1 + 2) MP7'.1 July 18, July 24
MP8 BBa_E0840 in pSB1A2 gfp tri-part; strong rbs (clones 1 + 2) July 18
MP9 BBa_J61047 in pSB1A2 Cre ORF (clones 1 + 2) MP9'.1, MP9'.2 July 18, July 24
MP10 BBa_I0500 in pSB2K3 AraC pBAD (clones 1 + 2) July 20
MP11 BBa_pJ23107 in p61002 Medium constitutive promoter in J61002 (clones 3 + 4) MP11'.3 July 20, July 23
MP12 BBa_P1003 in pSB1A2 KanR July 27
MP13 BBa_J61002 ptet-mRFP MP13.1, MP13.2 August 14
MP14 BBa_J61025 Medium constitutive promoter in J61002 (clones 1 + 2) MP14.1, MP14.2 August 14
L1 BBa_I0500 in J61002 construct with pBad promotor regulating expression of mRFP L1".1, L1".2 August 14
AttB MP15 August 14
AttP Finished MP16 July 25, July 27, Aug 7
L2 Lox66DapAE.coli in pSB1A2 Insertion of lox66DapAE.coli construct as a biobrick in pSB1A2 plasmid

http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png

July 25
L6 BBa_B0030+BBa_J61047 strong RBS(B0030)-Cre ORF

http://partsregistry.org/images/partbypart/icon_rbs.png http://partsregistry.org/wiki/images/7/7b/Part_icon_cds.png

L6.1, L6.2, L6.3, L6".1, L6".2 July 25, July 27
L9 lox66 in pSB1A2 July 25, July 30
L10 lox71 in pSB1A2 July 25, July 30
L12 lox71-gfp in pSB1A2 construction of the biobrick lox71-gfp

http://partsregistry.org/wiki/images/9/9f/Part_icon_dna.png http://partsregistry.org/images/partbypart/icon_reporter.png

L12.1 & L12.2 July 30
L26 araC/pbad>>RBS-Cre in J61002 ligation ( Aug.13) of RBS-Cre as a CDS behind pbad in J61002 L26.1 (no more) & L26.2 Aug. 17
L29 gfptripart-lox66 pSB1A2 ligation ( Aug.13) of gfp-tripart in front of lox66 L29.1 & L29.2 Aug. 17
L30 dapAp>>mRFP in J61002 dapAp promotor placed( Aug.13) upstream of mRFP J61002 L30A1 & L30A2 Aug. 17
L31 lox71-B0015(T) in pSB1A2 L31.1 & L31.3 Aug. 20
L32 B0015(T)-lox66 in pSB1A2 L32.1 & L32.2 Aug. 20
L34 pTet>>lox71-gfptripart in pSB1A2 L34.1 & L34.2 Aug. 20
L35 araC/pBad>>lox71-gfptripart in pSB1A2 il manque la seq. de 740-1410 L35.1 & L35.2 Aug. 20
L36 lox66-mRFP L36.1 & L36.2 Aug. 20
L37 lox66-RBS-dapAcoli L37.1 & L37.2 Aug. 20
L39 RBS-dapAcoli cloned in pSB1A2 L39.1 & L39.2 Aug. 20
L46 pTet>>lox71-B0015(T) in pSB1A2 L46.1 Sep. 18
L53 B0015(T)-lox66-RBSDapAcoli in pSB1A2 L53.7 Sep. 18
L56 B0015(T)-lox66-RBSdapAsubtilis in pSB1A2 L56.3 (attention, seq a partiellement foiré, refaire seq) Sep. 18
L59 pTet>>lox71-gfptripart-lox66-mRFP pSB1A2 L59.6 (attention, seq avec O40 a foiré, mais les extrémités sont bonnes, je pense que c ok car les colonies sont vertes) Sep. 18
L64 RBSdapAsubtilis cloned in pSB1A2 L564.1 & L64.2 Sep. 18
MPT1 Lox66-DapAColi pTOPO P5 MPT1.1, MPT1.2 Aug 7
MPT3 Lox66-DapASubtilis pTOPO P6 MPT3.2, MPT3.1 Aug 7
MPT4 RBS-DapAColi pTOPO P8 MPT4.1, .2, .3, .4& .5 Aug 7
MPT5 RBS-DapASubtilis pTOPO P9 MPT5.1, .2 &.3 Aug 7
MP15 pKD46 Oct 2

Digestion Products


PURPLE = Vector
GREEN = Insert

Digestion Products
Number Product Name Matrix Name Enzyme 1 Enzyme 2 Size Description Product Left []ng/µL Date
D1 pJ23100 pJ23100 Spe1 Pst1 2096 Vector with the J23100 promoter ready for insertion of a RBS/CDS insert Ø (24/09/07) 34 July 17
D2 mRFP pJ23100 Spe1 Pst1 887 27 July 17
D3 pJ23107 pJ23107 Spe1 Pst1 2096 Vector with the J23107 promoter ready for insertion of a RBS/CDS insert Ø (24/09/07) 32 July 17
D4 mRFP pJ23107 Spe1 Pst1 887 25 July 17
D5 pJ23100 pJ23100 EcoR1 Spe1 2925 The vector with the mRFP gene ready for insertion of a promoter. We are not sure the digestion worked properly. 28&29 July 17
D6 pJ23107 pJ23107 EcoR1 Spe1 2925 The vector with the mRFP gene ready for insertion of a promoter. We are not sure the digestion worked properly. 31&27 July 17
D7 Lox66 DapA E.coli Lox66 DapA E.coli (PCR product) EcoR1 Pst1 977 24 July 17
D8 Lox66 DapA E.coli Lox66 DapA E.coli (PCR product) Xba1 Pst1 962 44 July 17
D9 pJ23100 ES vector pJ23100 (MP1.1 & MP1.2) EcoI SpeI 2925 Vector with mRFP RBS/CDS ready for insertion promoter (fw-insert) 80&43 July 21
D10 pJ23100 promoter fw-insert pJ23100 (MP1.1 & MP1.2) EcoI SpeI 58 J23100 promoter ready for use as a fw-insert 35&28 July 21
D11 pJ23107 ES vector pJ23107 (MP2.1, MP2.2, MP2.3 & MP2.4) EcoRI SpeI 2925 Vector with mRFP RBS/CDS ready for insertion of a fw-insert promoter 49,35,28&41 July 21
D12 pJ23107 promoter fw-insert pJ23107 (MP2.1, MP2.2, MP2.3 & MP2.4) EcoRI SpeI 58 J23107 promoter ready for usage as a fw-insert 23,25,32&32 July 21
D13 mRFP pJ23107 (MP2.2, MP2.3 & MP2.4) SpeI PstI 887 mRFP RBS/CDS/T sequence, can it be used as a backward insert? => yes but you will not be able to add any biobrick after it 21&41 July 21
D14 Cre ORF J61047 MP9.1 & MP9.2 XbaI PstI 1063 Cre ORF extracted for usage as a bw-insert Ø D14.1 38&41 July 21
D15 B0030 bw-vector B0030 MP5.1 & MP5.2 SpeI PstI 2076 bacward vector for cloning a CDS bw-insert after the B0030 RBS 87&42 July 21
D18 BB dig. lox66DapAE.coli lox66-DapAE.coli P5 PCR product EcoRI PstI ~960 PCR product digested for cloning as a BioBrick Ø 44 July 21
D19 BB dig. lox66DapAsubtilis lox66-DapAsubtilis P6 PCR product EcoRI PstI ~960 PCR product digested for cloning as a BioBrick Ø July 21
D19' BB dig. lox66DapAsubtilis lox66-DapAsubtilis P6' PCR product EcoRI PstI ~960 PCR product digested for cloning as a BioBrick 16 July 30
D20 bw-insert lox66-DapAE.coli lox66-DapAE.coli P5 PCR product XbaI PstI ~960 PCR product digested for usage as a backward insert Ø July 21
D21 bw-insert lox66-DapAsubtilis lox66-DapAsubtilis P6 PCR product XbaI PstI ~960 PCR product digested for usage as a backward insert Ø July 21
D21' bw-insert lox66-DapAsubtilis lox66-DapAsubtilis P6' PCR product XbaI PstI ~960 PCR product digested for cloning as a BI 34 July 30
D22 pSB1A2 open vector BBa_J61047 (MP9.1 & MP9.2) EcoI PstI Ø D22.1 Open pSB1A2 vector for BioBrick EcoRI/PstI insertio Ø (24/09/07) D22.2:59 July 23, Aug. 13
D23 AraC/pBad promoter fw-insert BBa_I0500 (MP4.1 & MP4.2) EcoI SpeI 1233 AraC/pBad promoter ready for use as a forward insert Ø D23.1 93&53 July 23
D24 eCFP bw-insert BBa_E0422 (MP6.1 & MP6.2) XbaI PstI eCFP (RBS-OFR_LVA-T) ready for use as a backward insert 56&55 July 23
D26 gfp-tripart BBa_E0840 (MP8.1 & MP8.2) XbaI PstI gfp-tri part (strongRBS-ORF-T) ready for use as a backward insert 47&47 July 23
D27 lox71-ftsZ bw-insert lox71-ftsZ PCR product P7 XbaI PstI lox71-ftsZ(unmutated) ready for use as a backward insert Ø July 23
D27' lox71-ftsZ bw-insert lox71-ftsZ PCR product P7 XbaI PstI lox71-ftsZ(unmutated) ready for use as a backward insert 24 July 30
D28 RBS-DapAColi BB P8 (RBS-DapAColi) EcoRI PstI ~950 Biobrick 54 July 25
D29 RBS-DapAColi P8 (RBS-DapAColi) XbaI PstI ~950 Backward insert 44 July 25
D30 RBS-DapASubtilis BB P9 (RBS-DapASubtilis) EcoRI PstI ~950 Biobrick 72 July 25
D31 RBS-DapASubtilis P9 (RBS-DapASubtilis) XbaI PstI ~950 Backward insert 51 July 25
D32 Lox71-FtsA-FtsZ P10 Lox71-FtsA-FtsZ XbaI PstI ~2600 Backward insert - EcoRI site still presents : we can't use EcoRI 47 July 25
D33 RBS(B0030)CreORF L6' (clone 1 & 2) XbaI PstI RBS-Cre digestion for use as a BI 29&27 July 30, August 13
D34 lox66 L9' lox66 SpeI PstI Biobrick plasmid carrying lox66 digested as a BV 18 July 30
D35 lox71 L10' lox71 SpeI PstI Biobrick plasmid carrying lox71 digested as a BV July 30, August 13
D36 lox71-KmR-lox66 P11 lox71-KmR-lox66 EcoRI PstI lox71-KmR-lox66 digested as a biobrick insert July 30
D37 lox71-gfp(tripart) BI L12.1 & L12.2 miniprep products (from 30/07 ligation reaction) XbaI PstI lox71_RBS-gfp-double terminator extracted as a backward insert Aug. 6
D38 BB-insert lox66-DapAE.coli lox66-DapAE.coli in pTOPO (P5 PCR product)(clones 1+2) EcoRI PstI lox66-dapAE.coli extracted from pTOPO plasmid as a BB insert Aug. 6
D39' BI-insert lox66-DapAE.coli lox66-DapAE.coli in pTOPO (P5 PCR product)(clones 1+2) XbaI PstI lox66-dapAcoli extracted for usage as a backward insert Aug. 7
D40 BB-insert lox71-KmR-lox66 L25.1 miniprep product (from ligation rxn of 30/07 EcoRI PstI lox71-KmR-lox66 construct extracted for clonong as a biobrick Aug. 6
D41 araC/pBad-J61002 plasmid SP dig. L1.2, L1".1 & L1".2 miniprep product SpeI PstI J61002 plasmid containing araC/pBad promotor digested for insertion of a BI CDS Aug. 13
D42 Lox66 FV L9 EcoRI XbaI pSB1A2-lox66 as a Forward vector August 13
D43 gfp-tripart FI MP8.1 EcoRI SpeI gfp-tri part (strongRBS-ORF-T) ready for use as a forward insert August 13
D44 B0015 terminator BI B0015 in pSB1AK3 (MP3.3 Aug.14) XbaI Pst1 terminator B0015 extracted for use as a BI August 16
D45 B0015 terminator FI B0015 in pSB1AK3 (MP3.4 Aug.14) EcoRI SpeI terminator B0015 extracted for use as a FI August 16
D46 B0015 BV B0015 in pSB1AK3 (MP3.3 Aug.14) SpeI Pst1 B0015 in pSB1AK3 plasmid extracted for use as a BV August 16
D47 pSB1A2 XP dig. pSB1A2 carrying Cre ORF (MP9.1) XbaI PstI pSB1A2 open vector ready for cloning of BI August 16
D48 pSB1A2 ES dig. pSB1A2 carrying Cre ORF (MP9.2) EcoRI SpeI pSB1A2 open vector ready for cloning of FI August 16
D49 lox71-gfptripart BI lox71-gfptripart in pSB1A2 (L12.2) XbaI Pst1 lox71-gfptripart extracted for use as a BI August 16
D50 BBa_J61002 pTet SP dig. BBa_J61002 (MP13.1 Aug. 14) SpeI PstI BBa_J61002 pTet promoter vector ready for insertion of CDS as a BI August 16
D51 RBS-dapAcoli BBinsert pTOPO of RBS-dapAcoli (MPT4.3) EcoRI Pst1 RBS-dapAcoli extracted for cloning as a biobrick August 16
D52 RBS-dapAcoli BI pTOPO of RBS-dapAcoli (MPT4.3) XbaI Pst1 RBS-dapAcoli extracted for use as a BI August 16
D53 RBS-dapAsubtilis BBinsert pTOPO of RBS-dapAsubtilis (MPT5.1) EcoRI Pst1 RBS-dapAsubtilis extracted for cloning as a biobrick August 16
D54 RBS-dapAsubtilis BI pTOPO of RBS-dapAsubtilis (MPT5.1) XbaI Pst1 RBS-dapAsubtilis extracted for use as a BI Ø (24/09/07) August 16
D55 RBS-dapAsubtilis FI pTOPO of RBS-dapAsubtilis (MPT5.2) EcoRI SpeI RBS-dapAsubtilis extracted for use as a FI August 16
D57 lox71-B0015(T) BI (L31.1, L31.3) lox71-B0015(T) pSB1A2 XbaI Pst1 lox71-B0015(T) extracted for use as a BI pSB1A2 Sep. 14
D58 B0015(T)-lox66 BV (L32.1, L32.2) B0015(T)-lox66 BV SpeI PstI B0015(T)-lox66 for use as a BV pSB1A2 Sep. 14
D59 pTet>>lox71-gfptripart BI (L34.1, L34.2) pTet>>lox71-gfptripart BBa_J61002 XbaI PstI pTet>>lox71-gfptripart for use as a BI Sep. 14
D61 lox66-mRFP FV (L36.1, L36.2) lox66-mRFP EcoRI XbaI lox66-mRFP part (strongRBS-ORF-T) ready for use as a forward vector Sep. 14
D62 lox66-RBSDapAcoli BI (L37.1, L37.2) lox66-RBSDapAcoli XbaI PstI lox66-RBSDapAcoli ready for use as a Backward insert Sep. 14
D63 RBSDapAcoli BI (L39.1, L39.2) RBSDapAcoli XbaI PstI RBSDapAcoli BI (for insertion behind T-lox66) Sep. 14
D64 FRT-CmR-FRT BBa_J61025 BV (MP14.1) BBa_J61025 SpeI PstI FRT-CmR-FRT BBa_J61025 (for backward introduction of a casette for Wanner K insertion) Sep. 14
D65 FRT-CmR-FRT BBa_J61025 FV (MP14.1) BBa_J61025 EcoR XbaI FRT-CmR-FRT BBa_J61025 (for forward introduction of a casette for Wanner K insertion) Sep. 14
D68 attB BV (MP15.1) SpeI PstI vector containing attB for use as a BV Sep. 14
D69 gfptripart-lox66 BV (L29.1) gfptripart-lox66 pSB1A2 SpeI PstI Sep. 14
D70 pTet>>lox71-gfptripart FI (L34.1, L34.2) pTet>>lox71-gfptripart BBa_J61002 EcoRI SpeI pTet>>lox71-gfptripart for use as a FI Sep. 14
D71 araC/pBad>>lox71-gfptripart FI L35.1 araC/pBad>>lox71-gfptripart BBa_J61002 EcoRI SpeI araC/pBad>>lox71-gfptripart for use as a FI Ø (24/09/07) Sep. 14 Sep. 17
D72 pSB2K3 BBvector MP16 pSB2K3::attP EcoRI PstI Low copy number plasmid, BB vector Sep. 17
D73 pSB2K3 ES open vector MP16 pSB2K3::attP EcoRI SpeI Comparison of digestion efficiency between Pst and Spe in NEB2 Sep. 17
D76 DGAT BB P14 DGAT EcoRI PstI DGAT Ø (24/09/07) Sep. 17
D77 araC/pBad>>lox71-gfptripart BI D60 (.1 & .2) digestion product FspI AhdI in D60, the extracted fragment & the open plasmid have the same size, it is thus necessary to remove the plasmid by digestion Sep. 17
D78 araC/pBad>>lox71-gfptripart FI D71 (.1 & .2) digestion product FspI AhdI in D71, the extracted fragment & the open plasmid have the same size, it is thus necessary to remove the plasmid by digestion Sep. 17

Unexpected digestion results

Unexpected results
Number Matrix Name Enzyme 1 Enzyme 2 Expected Size Description Date
D16 pSB1AK3::B0015 (MP3.1 & MP3.2) EcoR1 Pst1 ~3000 We have one band at ~2000. Furthermore, the strain which is suppose to be resistant to Kanamycin is not. We suspect that the part is in the pSB1A2 instead of the pSB1AK3 July 21
D17 pSB1A2::E0422 (MP6.1 & MP6.2) EcoRI PstI ~2040 + ~920 3 bands July 21
D25 BBa_E0421 (MP7.1 & MP7.2) PoPs to GFP converter XbaI PstI July 23
D66 (MP12.1, MP12.2) BBa_P1003 in pSB1A2 EcoR XbaI KanR BBa_J61025 FV (for forward introduction of a casette for Wanner K insertion). The strange thing is that this digestion produces 2 visible products: ~1000 bp & ~2500 bp!!! Sep.17, Sep.27

Strains (glycerol stock)

Strains glycerol stock
Number Name Description Verified clone Date
S1 MG1655 Standard E.Coli strain
S2 Acinetobacter ADP1 Strain used for TG coloration test with Red Nile
S3 DH5alpha pKs::DGAT Enzyme catalysing the transormation of DAG into TG
S4 BBa_pJ23100 Strong constitutive promoter in J61002 (clones 1 + 2)
S5 BBa_B0015 Double terminator (clones 1 + 2)
S6 BBa_pJ23107 Medium constitutive promoter in J61002 (clones 1 + 2) July 19
S7 BBa_B0030 Strong RBS (clones 1 + 2) July 19
S8 BBa_E0422 ECFP (RBS+LVA+Term) (clones 1 + 2) July 19
S9 BBa_E0241 PoPs to GFP converter (clones 1 + 2) July 19
S10 BBa_E0840 gfp tri-part; strong rbs (clones 1 + 2) July 19
S11 BBa_J61047 Cre ORF (clones 1 + 2) July 19
S12 W121 05/07/07
S13 BBa_I0500 pBAD AraC (clone 1+2) July 20
S14 BBa_pJ23107 Medium constitutive promoter in J61002 (clone 3+4) July 20
S15 BBa_P1003 Kanamycin resistance cassette (clone 1+2) July 20
S16 J61002::I0500 AraC pBad regulating mRFP expression in pJ61002 (L1) L1.2, L1.3 July 27
S17 pSB1A2::Lox66-DapA Coli Lox66-DapA Coli cloned as a biobrick (L2) July 27
S18 pSB1A2::B0030 + J61047 RBS(B0030)-Cre ORF (L6) (clones 1+2) July 27
S19 pSB1A2::BBa_J37026 Lox66 (L9)(1 clone) July 27
S20 pSB1A2::BBa_J37028 Lox71 (L10)(1 clone) July 27
S21 pSB1A2::BBa_J37028+BBa_E0840 Lox71-gfp tri-part (L12) L12.2 ok ?
S22 pSB1A2::Lox71-kmR-lox66 Lox71-kmR-lox66 (L25) sequence not good (thrown away) ?
S23 pTopo::P5 Lox66-DapAColi ?
S24 pTopo::P6 Lox66-DapASubtilis ?
S25 pTopo::P8 RBS-DapAColi MPT4.3, MPT4.5 ?
S26 pTopo::P9 RBS-DapASubtilis ?
S27 J61002 August 14
S28 J61025 FRT-Cm-FRT August 14
S29 L26 Insertion of RBS-Cre behind araC/pBad August 13
S30 L27 Insertion of ftsZ behind araC/pBad August 13
S31 L28 Insertion of ftsAZ behind araC/pBad August 13
S32 L29 Insertion of gfp-tripart in front of lox66 August 13
S33A L30A cloning dapAp promotor in J61002 August 13
S33B L30B cloning dapAp promotor in J61002 August 13
S34 L31 Insertion of B0015 terminator behind lox71 August 22
S35 L32 Insertion of B0015 terminator in front of lox66 August 22
S36 L34 pTet>>lox71-gfptripart August 22
S37 L35 araC/pBad>>lox71-gfptripart August 22
S38 L36 lox66-mRFP August 22
S39 L37 lox66-RBS-dapAcoli BI August 22
S40 L39 RBS-dapAcoli cloned in pSB1A2 August 22
S41 L40 RBS-dapAsubtilis cloned in pSB1A2 August 22
S42 L41 lox71-ftsZ cloned in pSB1A2 August 22
S43 L42 lox71-ftsA-Z cloned in pSB1A2 August 22
S44 L43 RBS-dapAsubtilis cloned in pSB1A2 August 22
S45A L30A DapAp>>mRFP within W121 strain August 25
S45B L30B DapAp>>mRFP within W121 strain August 25
S48 pKD13 FRT-KanR-FRT; Wanner plasmid Sep. 18
S49 pKD46 Red recombination system; Wanner plasmid Sep. 13
S50 L30 plasmid in FR781 dapAp>>mRFP plasmid Sep. 28


  • p2C (CFP fluorescent)
  • p2Y (YFP fluorescent)
  • FtsZTS strains
    • MG1655 FtsZ -TS 84
    • 121.1
    • 121.2
    • 121.3
    • 121.4
    • 121.5
    • 121.6
    • 129
    • 1.129
    • DCR 14.1
    • DCR 14.2
    • DCR 14.3
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