Paris/Freezer
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(227 intermediate revisions not shown) | |||
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|- style="background: #ccccff; text-align: center;" | |- style="background: #ccccff; text-align: center;" | ||
|width=5%| Number | |width=5%| Number | ||
- | |width= | + | |width=10%| Name |
+ | |width=12.5%| Oligo F | ||
+ | |width=12.5%| Oligo R | ||
+ | |width=10%| Matrix | ||
|width=5%| Length | |width=5%| Length | ||
- | |width= | + | |width=25%| Comments |
|width=5%| Product Left ? | |width=5%| Product Left ? | ||
+ | |width=5%| []ng/µL | ||
|width=5%| Date | |width=5%| Date | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |P1 |
|Lox71-FtsA-FtsZ-1 | |Lox71-FtsA-FtsZ-1 | ||
+ | |O3 Lox71-FtsA-F | ||
+ | |O4 DEcoR1-FtsZ-R | ||
+ | |MG1655 | ||
|1537 | |1537 | ||
| | | | ||
+ | |Ø | ||
| | | | ||
| | | | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |P2 |
|FtsZ-2 | |FtsZ-2 | ||
+ | |O5 DEcoR1-FtsZ-F | ||
+ | |O2 FtsZ-R | ||
+ | |MG1655 | ||
|1088 | |1088 | ||
| | | | ||
+ | |Ø | ||
| | | | ||
| | | | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | 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;" | | + | | 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;" | | + | | style="background: #ccffcc;" |P5 |
|Lox66-DapAColi | |Lox66-DapAColi | ||
+ | |O6 Lox66-DapAColi-F | ||
+ | |O7 DapAColi-R | ||
+ | |MG1655 | ||
|985 | |985 | ||
| | | | ||
+ | |Ø | ||
| | | | ||
- | | | + | |[[Paris/July 17| July 17]], [[Paris/July 30| July 30]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |P6 |
|Lox66-DapASubtilis | |Lox66-DapASubtilis | ||
+ | |O8 Lox66-DapASubt-F | ||
+ | |O9 DapASubt-R | ||
+ | |B.Subtilis 168 | ||
|979 | |979 | ||
| | | | ||
| | | | ||
- | |17/ | + | | |
+ | |[[Paris/July 17| July 17]],[[Paris/July 27| July 27]],[[Paris/July 30| July 30]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |P7 |
|Lox71-FtsZ | |Lox71-FtsZ | ||
+ | | O1 Lox71-FtsZ-F | ||
+ | | O2 FtsZ-R | ||
+ | |MG1655 | ||
|1260 | |1260 | ||
- | | | + | |EcoRI site in FtsZ not mutated |
| | | | ||
- | |17/ | + | | |
+ | |[[Paris/July 17| July 17]],[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |P8 | ||
+ | |RBS-DapAColi | ||
+ | | O20 RBS-DapAColi-F | ||
+ | | O7 DapAColi-R | ||
+ | | MG1655 | ||
+ | |952 | ||
+ | |B0030-DapAColi | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 23| July 23]],[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |P9 | ||
+ | |RBS-DapASubtilis | ||
+ | | O21 RBS-DapASubt-F | ||
+ | | O9 DapASubt-R | ||
+ | |B.Subtilis 168 | ||
+ | |946 | ||
+ | |B0030-DapASubtilis | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 23| July 23]], [[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |P10 | ||
+ | |Lox71-FtsA-FtsZ | ||
+ | |O3 Lox71-FtsA-F | ||
+ | |O2 FtsZ-R | ||
+ | |MG1655 | ||
+ | |2580 | ||
+ | |EcoRI site in FtsZ not mutated | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 23| July 23]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |P11 | ||
+ | |Lox71-KmR-Lox66 | ||
+ | | O10 Lox71-KmR-F | ||
+ | | O11 Lox66-KmR-R | ||
+ | | MP12 | ||
+ | |1084 | ||
+ | | | ||
+ | | | ||
+ | | | ||
+ | |[[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_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 == | ||
Line 68: | Line 207: | ||
|width=5%| Number | |width=5%| Number | ||
|width=25%| Name | |width=25%| Name | ||
- | |width= | + | |width=50%| Description |
- | |width= | + | |width=15%| Products in use |
|width=5%| Date | |width=5%| Date | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP1 |
|<bbpart>pJ23100</bbpart> in <bbpart>J61002</bbpart> | |<bbpart>pJ23100</bbpart> in <bbpart>J61002</bbpart> | ||
- | |Strong constitutive promoter in J61002 (clones 1 + 2) | + | |Strong constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23100 SpecifiedComponents</partinfo> |
| | | | ||
- | | | + | | [[Paris/July 15| July 15]], [[Paris/August 17| Aug. 17]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP2 |
- | |<bbpart>pJ23107</bbpart> in <bbpart>J61002</bbpart> | + | |<bbpart>pJ23107</bbpart> in <bbpart>J61002</bbpart> |
- | |Medium constitutive promoter in J61002 (clones 1 + 2) | + | |Medium constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23107 SpecifiedComponents</partinfo> |
- | | | + | |MP2.2 |
- | | 15/ | + | | [[Paris/July 15| July 15]], [[Paris/August 17| Aug. 17]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP3 |
|<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> | |<bbpart>B0015</bbpart> in <bbpart>pSB1AK3</bbpart> | ||
- | |Double terminator (clones 1 + 2) | + | |Double terminator (clones 1 + 2) <partinfo>B0015 SpecifiedComponents</partinfo> |
- | | | + | |MP3.3, MP3.4 |
- | | | + | | [[Paris/August 14| August 14]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP4 |
|<bbpart>I0500</bbpart> in <bbpart>pSB2K3</bbpart> | |<bbpart>I0500</bbpart> in <bbpart>pSB2K3</bbpart> | ||
- | |AraC pBAD (clones 1 + 2) | + | |AraC pBAD (clones 1 + 2) <partinfo>I0500 SpecifiedComponents</partinfo> |
| | | | ||
- | | | + | | [[Paris/July 16| July 16]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP5 |
|<bbpart>BBa_B0030</bbpart> in <bbpart>pSB1A2</bbpart> | |<bbpart>BBa_B0030</bbpart> in <bbpart>pSB1A2</bbpart> | ||
- | |Strong RBS (clones 1 + 2) | + | |Strong RBS (clones 1 + 2) <partinfo>B0030 SpecifiedComponents</partinfo> |
| | | | ||
- | | | + | | [[Paris/July 18| July 18]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP6 |
|<bbpart>BBa_E0422</bbpart> in <bbpart>pSB1A2</bbpart> | |<bbpart>BBa_E0422</bbpart> in <bbpart>pSB1A2</bbpart> | ||
- | |ECFP (RBS+LVA+Term) (clones 1 + 2) | + | |ECFP (RBS+LVA+Term) (clones 1 + 2) <partinfo>E0422 SpecifiedComponents</partinfo> |
- | | | + | |MP6'.1, MP6'.2 |
- | | 18/ | + | | [[Paris/July 18| July 18]], [[Paris/July 24| July 24]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP7 |
|<bbpart>BBa_E0241</bbpart> in <bbpart>pSB1A2</bbpart> | |<bbpart>BBa_E0241</bbpart> in <bbpart>pSB1A2</bbpart> | ||
- | |PoPs to GFP converter (clones 1 + 2) | + | |PoPs to GFP converter (clones 1 + 2) <partinfo>BBa_E0241 DeepComponents</partinfo> |
- | | | + | |MP7'.1 |
- | | 18/ | + | | [[Paris/July 18| July 18]], [[Paris/July 24| July 24]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP8 |
|<bbpart>BBa_E0840</bbpart> in <bbpart>pSB1A2</bbpart> | |<bbpart>BBa_E0840</bbpart> in <bbpart>pSB1A2</bbpart> | ||
- | |gfp tri-part; strong rbs (clones 1 + 2) | + | |gfp tri-part; strong rbs (clones 1 + 2) <partinfo>E0840 SpecifiedComponents</partinfo> |
| | | | ||
- | | | + | | [[Paris/July 18| July 18]] |
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |MP9 |
|<bbpart>BBa_J61047</bbpart> in <bbpart>pSB1A2</bbpart> | |<bbpart>BBa_J61047</bbpart> in <bbpart>pSB1A2</bbpart> | ||
- | |Cre ORF (clones 1 + 2) | + | |Cre ORF (clones 1 + 2) <partinfo>J61047 SpecifiedComponents</partinfo> |
+ | |MP9'.1, MP9'.2 | ||
+ | | [[Paris/July 18| July 18]], [[Paris/July 24| July 24]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MP10 | ||
+ | |<bbpart>BBa_I0500</bbpart> in <bbpart>pSB2K3</bbpart> | ||
+ | |AraC pBAD (clones 1 + 2) <partinfo>I0500 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | | [[Paris/July 20| July 20]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MP11 | ||
+ | |<bbpart>BBa_pJ23107</bbpart> in <bbpart>p61002</bbpart> | ||
+ | |Medium constitutive promoter in J61002 (clones 3 + 4) <partinfo>pJ23107 SpecifiedComponents</partinfo> | ||
+ | |MP11'.3 | ||
+ | | [[Paris/July 20| July 20]], [[Paris/July 23| July 23]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MP12 | ||
+ | |<bbpart>BBa_P1003</bbpart> in <bbpart>pSB1A2</bbpart> | ||
+ | |KanR <partinfo>P1003 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | | [[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: #ccffcc;" |L1 | ||
+ | |<bbpart>BBa_I0500</bbpart> in <bbpart>J61002</bbpart> | ||
+ | |construct with pBad promotor regulating expression of mRFP <partinfo>I0500 SpecifiedComponents</partinfo> | ||
+ | |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]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |L2 | ||
+ | |Lox66DapAE.coli in <bbpart>pSB1A2</bbpart> | ||
+ | |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 | ||
+ | | | ||
+ | | [[Paris/July 25| July 25]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |L6 | ||
+ | |<bbpart>BBa_B0030</bbpart>+<bbpart>BBa_J61047</bbpart> | ||
+ | |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 | ||
+ | | [[Paris/July 25| July 25]], [[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |L9'' | ||
+ | |lox66 in <bbpart>pSB1A2</bbpart> | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 25| July 25]], [[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |L10'' | ||
+ | |lox71 in <bbpart>pSB1A2</bbpart> | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 25| July 25]], [[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |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 | ||
+ | |[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |L26 | ||
+ | |araC/pbad>>RBS-Cre in <bbpart>J61002</bbpart> | ||
+ | |ligation ([[Paris/August 13| Aug.13]]) of RBS-Cre as a CDS behind pbad in <bbpart>J61002</bbpart> | ||
+ | |L26.1 (no more) & L26.2 | ||
+ | |[[Paris/August 17| Aug. 17]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |L29 | ||
+ | |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: #ccffcc;" |MPT1 | ||
+ | |Lox66-DapAColi | ||
+ | |pTOPO P5 | ||
+ | |MPT1.1, MPT1.2 | ||
+ | |[[Paris/August 7| Aug 7]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MPT3 | ||
+ | |Lox66-DapASubtilis | ||
+ | |pTOPO P6 | ||
+ | |MPT3.2, MPT3.1 | ||
+ | |[[Paris/August 7| Aug 7]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MPT4 | ||
+ | |RBS-DapAColi | ||
+ | |pTOPO P8 | ||
+ | |MPT4.1, .2, .3, .4& .5 | ||
+ | |[[Paris/August 7| Aug 7]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MPT5 | ||
+ | |RBS-DapASubtilis | ||
+ | |pTOPO P9 | ||
+ | |MPT5.1, .2 &.3 | ||
+ | |[[Paris/August 7| Aug 7]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |MP15 | ||
+ | |pKD46 | ||
+ | | | ||
| | | | ||
- | | | + | |[[Paris/October 2| Oct 2]] |
|} | |} | ||
== Digestion Products == | == Digestion Products == | ||
+ | <br> | ||
+ | <font color="#FF99FF" size="4">PURPLE = Vector </font><br> | ||
+ | <font color="#ccffcc" size="4">GREEN = Insert </font><br> | ||
{| | {| | ||
|- style="background: #ccccff;" | |- style="background: #ccccff;" | ||
Line 137: | Line 483: | ||
|width=5%| Enzyme 1 | |width=5%| Enzyme 1 | ||
|width=5%| Enzyme 2 | |width=5%| Enzyme 2 | ||
- | |width= | + | |width=5%| Size |
+ | |width=30%| Description | ||
+ | |width=5%| Product Left | ||
+ | |width=5%| []ng/µL | ||
|width=5%| Date | |width=5%| Date | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: # | + | | style="background: #FF99FF;" |D1 |
+ | |<bbpart>pJ23100</bbpart> | ||
|<bbpart>pJ23100</bbpart> | |<bbpart>pJ23100</bbpart> | ||
- | |||
- | |||
|Spe1 | |Spe1 | ||
- | | | + | |Pst1 |
- | | | + | |2096 |
+ | |Vector with the J23100 promoter ready for insertion of a RBS/CDS insert | ||
+ | |Ø (24/09/07) | ||
+ | |34 | ||
+ | |[[Paris/July 17| July 17]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
| style="background: #ccffcc;" |D2 | | style="background: #ccffcc;" |D2 | ||
+ | |<bbpart>mRFP dblTer</bbpart> | ||
|<bbpart>pJ23100</bbpart> | |<bbpart>pJ23100</bbpart> | ||
- | |||
|Spe1 | |Spe1 | ||
|Pst1 | |Pst1 | ||
+ | |887 | ||
| | | | ||
| | | | ||
+ | |27 | ||
+ | |[[Paris/July 17| July 17]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: # | + | | style="background: #FF99FF;" |D3 |
|<bbpart>pJ23107</bbpart> | |<bbpart>pJ23107</bbpart> | ||
+ | |<bbpart>pJ23107</bbpart> | ||
+ | |Spe1 | ||
+ | |Pst1 | ||
+ | |2096 | ||
+ | |Vector with the J23107 promoter ready for insertion of a RBS/CDS insert | ||
+ | |Ø (24/09/07) | ||
+ | |32 | ||
+ | |[[Paris/July 17| July 17]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D4 | ||
+ | |<bbpart>mRFP dblTer</bbpart> | ||
+ | |<bbpart>pJ23107</bbpart> | ||
+ | |Spe1 | ||
+ | |Pst1 | ||
+ | |887 | ||
| | | | ||
+ | | | ||
+ | |25 | ||
+ | |[[Paris/July 17| July 17]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D5 | ||
+ | |<bbpart>pJ23100</bbpart> | ||
+ | |<bbpart>pJ23100</bbpart> | ||
|EcoR1 | |EcoR1 | ||
|Spe1 | |Spe1 | ||
+ | |2925 | ||
+ | |The vector with the mRFP gene ready for insertion of a promoter. We are not sure the digestion worked properly. | ||
| | | | ||
- | | | + | |28&29 |
+ | |[[Paris/July 17| July 17]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: # | + | | style="background: #FF99FF;" |D6 |
|<bbpart>pJ23107</bbpart> | |<bbpart>pJ23107</bbpart> | ||
- | | | + | |<bbpart>pJ23107</bbpart> |
+ | |EcoR1 | ||
|Spe1 | |Spe1 | ||
+ | |2925 | ||
+ | |The vector with the mRFP gene ready for insertion of a promoter. We are not sure the digestion worked properly. | ||
+ | | | ||
+ | |31&27 | ||
+ | |[[Paris/July 17| July 17]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D7 | ||
+ | |Lox66 DapA E.coli | ||
+ | |Lox66 DapA E.coli (PCR product) | ||
+ | |EcoR1 | ||
|Pst1 | |Pst1 | ||
+ | |977 | ||
| | | | ||
| | | | ||
+ | |24 | ||
+ | |[[Paris/July 17| July 17]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |D8 |
- | | | + | |Lox66 DapA E.coli |
+ | |Lox66 DapA E.coli (PCR product) | ||
+ | |Xba1 | ||
+ | |Pst1 | ||
+ | |962 | ||
| | | | ||
- | | | + | | |
+ | |44 | ||
+ | |[[Paris/July 17| July 17]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D9 | ||
+ | |<bbpart>pJ23100</bbpart> ES vector | ||
+ | |<bbpart>pJ23100</bbpart> (MP1.1 & MP1.2) | ||
+ | |EcoI | ||
+ | |SpeI | ||
+ | |2925 | ||
+ | |Vector with mRFP RBS/CDS ready for insertion promoter (fw-insert) | ||
+ | | | ||
+ | |80&43 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D10 | ||
+ | |<bbpart>pJ23100</bbpart> promoter fw-insert | ||
+ | |<bbpart>pJ23100</bbpart> (MP1.1 & MP1.2) | ||
+ | |EcoI | ||
+ | |SpeI | ||
+ | |58 | ||
+ | |J23100 promoter ready for use as a fw-insert | ||
+ | | | ||
+ | |35&28 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D11 | ||
+ | |<bbpart>pJ23107</bbpart> ES vector | ||
+ | |<bbpart>pJ23107</bbpart> (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 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D12 | ||
+ | |<bbpart>pJ23107</bbpart> promoter fw-insert | ||
+ | |<bbpart>pJ23107</bbpart> (MP2.1, MP2.2, MP2.3 & MP2.4) | ||
+ | |EcoRI | ||
+ | |SpeI | ||
+ | |58 | ||
+ | |J23107 promoter ready for usage as a fw-insert | ||
+ | | | ||
+ | |23,25,32&32 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D13 | ||
+ | |<bbpart>mRFP</bbpart> | ||
+ | |<bbpart>pJ23107</bbpart> (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 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D14 | ||
+ | |Cre ORF | ||
+ | |<bbpart>J61047</bbpart> MP9.1 & MP9.2 | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | |1063 | ||
+ | |Cre ORF extracted for usage as a bw-insert | ||
+ | |Ø D14.1 | ||
+ | |38&41 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D15 | ||
+ | |<bbpart>B0030</bbpart> bw-vector | ||
+ | |<bbpart>B0030</bbpart> MP5.1 & MP5.2 | ||
+ | |SpeI | ||
+ | |PstI | ||
+ | |2076 | ||
+ | |bacward vector for cloning a CDS bw-insert after the B0030 RBS | ||
+ | | | ||
+ | |87&42 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D18 | ||
+ | |BB dig. lox66DapAE.coli | ||
+ | |lox66-DapAE.coli P5 PCR product | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | |~960 | ||
+ | |PCR product digested for cloning as a BioBrick | ||
+ | |Ø | ||
+ | |44 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D19 | ||
+ | |BB dig. lox66DapAsubtilis | ||
+ | |lox66-DapAsubtilis P6 PCR product | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | |~960 | ||
+ | |PCR product digested for cloning as a BioBrick | ||
+ | |Ø | ||
+ | | | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D19' | ||
+ | |BB dig. lox66DapAsubtilis | ||
+ | |lox66-DapAsubtilis P6' PCR product | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | |~960 | ||
+ | |PCR product digested for cloning as a BioBrick | ||
+ | | | ||
+ | |16 | ||
+ | |[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |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 | ||
+ | | | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D21 | ||
+ | |bw-insert lox66-DapAsubtilis | ||
+ | |lox66-DapAsubtilis P6 PCR product | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | |~960 | ||
+ | |PCR product digested for usage as a backward insert | ||
+ | |Ø | ||
+ | | | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D21' | ||
+ | |bw-insert lox66-DapAsubtilis | ||
+ | |lox66-DapAsubtilis P6' PCR product | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | |~960 | ||
+ | |PCR product digested for cloning as a BI | ||
+ | | | ||
+ | |34 | ||
+ | |[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D22 | ||
+ | |pSB1A2 open vector | ||
+ | |<bbpart>BBa_J61047</bbpart> (MP9.1 & MP9.2) | ||
+ | |EcoI | ||
+ | |PstI | ||
+ | |Ø D22.1 | ||
+ | |Open pSB1A2 vector for BioBrick EcoRI/PstI insertio | ||
+ | |Ø (24/09/07) | ||
+ | |D22.2:59 | ||
+ | |[[Paris/July 23| July 23]],[[Paris/August 13 | Aug. 13 ]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D23 | ||
+ | |AraC/pBad promoter fw-insert | ||
+ | |<bbpart>BBa_I0500</bbpart> (MP4.1 & MP4.2) | ||
+ | |EcoI | ||
+ | |SpeI | ||
+ | |1233 | ||
+ | |AraC/pBad promoter ready for use as a forward insert | ||
+ | |Ø D23.1 | ||
+ | |93&53 | ||
+ | |[[Paris/July 23| July 23]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D24 | ||
+ | |eCFP bw-insert | ||
+ | |<bbpart>BBa_E0422</bbpart> (MP6.1 & MP6.2) | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | |eCFP (RBS-OFR_LVA-T) ready for use as a backward insert | ||
+ | | | ||
+ | |56&55 | ||
+ | |[[Paris/July 23| July 23]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D26 | ||
+ | |gfp-tripart | ||
+ | |<bbpart>BBa_E0840</bbpart> (MP8.1 & MP8.2) | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | |gfp-tri part (strongRBS-ORF-T) ready for use as a backward insert | ||
+ | | | ||
+ | |47&47 | ||
+ | |[[Paris/July 23| July 23]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D27 | ||
+ | |lox71-ftsZ bw-insert | ||
+ | |lox71-ftsZ PCR product P7 | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | |lox71-ftsZ(unmutated) ready for use as a backward insert | ||
+ | |Ø | ||
+ | | | ||
+ | |[[Paris/July 23| July 23]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D27' | ||
+ | |lox71-ftsZ bw-insert | ||
+ | |lox71-ftsZ PCR product P7 | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | |lox71-ftsZ(unmutated) ready for use as a backward insert | ||
+ | | | ||
+ | |24 | ||
+ | |[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D28 | ||
+ | |RBS-DapAColi BB | ||
+ | |P8 (RBS-DapAColi) | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | |~950 | ||
+ | |Biobrick | ||
+ | | | ||
+ | |54 | ||
+ | |[[Paris/July 25| July 25]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D29 | ||
+ | |RBS-DapAColi | ||
+ | |P8 (RBS-DapAColi) | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | |~950 | ||
+ | |Backward insert | ||
+ | | | ||
+ | |44 | ||
+ | |[[Paris/July 25| July 25]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D30 | ||
+ | |RBS-DapASubtilis BB | ||
+ | |P9 (RBS-DapASubtilis) | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | |~950 | ||
+ | |Biobrick | ||
+ | | | ||
+ | |72 | ||
+ | |[[Paris/July 25| July 25]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D31 | ||
+ | |RBS-DapASubtilis | ||
+ | |P9 (RBS-DapASubtilis) | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | |~950 | ||
+ | |Backward insert | ||
+ | | | ||
+ | |51 | ||
+ | |[[Paris/July 25| July 25]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D32 | ||
+ | |Lox71-FtsA-FtsZ | ||
+ | |P10 Lox71-FtsA-FtsZ | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | |~2600 | ||
+ | |Backward insert - EcoRI site still presents : we can't use EcoRI | ||
+ | | | ||
+ | |47 | ||
+ | |[[Paris/July 25| July 25]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D33 | ||
+ | |RBS(B0030)CreORF | ||
+ | |L6' (clone 1 & 2) | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | |RBS-Cre digestion for use as a BI | ||
+ | | | ||
+ | |29&27 | ||
+ | |[[Paris/July 30| July 30]],[[Paris/August 13| August 13]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D34 | ||
+ | |lox66 | ||
+ | |L9' lox66 | ||
+ | |SpeI | ||
+ | |PstI | ||
+ | | | ||
+ | |Biobrick plasmid carrying lox66 digested as a BV | ||
+ | | | ||
+ | |18 | ||
+ | |[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D35 | ||
+ | |lox71 | ||
+ | |L10' lox71 | ||
+ | |SpeI | ||
+ | |PstI | ||
+ | | | ||
+ | |Biobrick plasmid carrying lox71 digested as a BV | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 30| July 30]],[[Paris/August 13| August 13]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D36 | ||
+ | |lox71-KmR-lox66 | ||
+ | |P11 lox71-KmR-lox66 | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | | | ||
+ | |lox71-KmR-lox66 digested as a biobrick insert | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/July 30| July 30]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D37 | ||
+ | |lox71-gfp(tripart) BI | ||
+ | |L12.1 & L12.2 miniprep products (from [[Paris/July 30| 30/07]] ligation reaction) | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | |lox71_RBS-gfp-double terminator extracted as a backward insert | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 6 | Aug. 6 ]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |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 | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 6| Aug. 6]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |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 | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 7 | Aug. 7 ]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D40 | ||
+ | |BB-insert lox71-KmR-lox66 | ||
+ | |L25.1 miniprep product (from ligation rxn of [[Paris/July 30| 30/07]] | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | | | ||
+ | |lox71-KmR-lox66 construct extracted for clonong as a biobrick | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 6 | Aug. 6 ]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |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 | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 13 | Aug. 13 ]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #FF99FF;" |D42 | ||
+ | |Lox66 FV | ||
+ | |L9 | ||
+ | |EcoRI | ||
+ | |XbaI | ||
+ | | | ||
+ | |pSB1A2-lox66 as a Forward vector | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 13| August 13]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D43 | ||
+ | |gfp-tripart FI | ||
+ | |MP8.1 | ||
+ | |EcoRI | ||
+ | |SpeI | ||
+ | | | ||
+ | |gfp-tri part (strongRBS-ORF-T) ready for use as a forward insert | ||
+ | | | ||
+ | | | ||
+ | |[[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 | |Pst1 | ||
| | | | ||
+ | |terminator B0015 extracted for use as a BI | ||
| | | | ||
+ | | | ||
+ | |[[Paris/August 16| August 16]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: #ccffcc;" | | + | | style="background: #ccffcc;" |D45 |
- | |<bbpart>pSB1AK3</bbpart> | + | |<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 | |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: #cccccc;" | ||
- | | style="background: # | + | | 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 | |Pst1 | ||
| | | | ||
+ | |lox71-gfptripart extracted for use as a BI | ||
| | | | ||
+ | | | ||
+ | |[[Paris/August 16| August 16]] | ||
|- style="background: #cccccc;" | |- style="background: #cccccc;" | ||
- | | style="background: # | + | | 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 | |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]] | ||
+ | |} | ||
+ | |||
+ | == Unexpected digestion results == | ||
+ | {| | ||
+ | |- style="background: #ccccff;" | ||
+ | ! colspan="5" style="background: #ccccff;" | Unexpected results | ||
+ | |- style="background: #ccccff; text-align: center;" | ||
+ | |width=5%| Number | ||
+ | |width=20%| Matrix Name | ||
+ | |width=5%| Enzyme 1 | ||
+ | |width=5%| Enzyme 2 | ||
+ | |width=5%| Expected Size | ||
+ | |width=55%| Description | ||
+ | |width=5%| Date | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D16 | ||
+ | |<bbpart>pSB1AK3</bbpart>::<bbpart>B0015</bbpart> (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 | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D17 | ||
+ | |<bbpart>pSB1A2</bbpart>::<bbpart>E0422</bbpart> (MP6.1 & MP6.2) | ||
+ | |EcoRI | ||
+ | |PstI | ||
+ | |~2040 + ~920 | ||
+ | |3 bands | ||
+ | |[[Paris/July 21| July 21]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |D25 | ||
+ | |<bbpart>BBa_E0421</bbpart> (MP7.1 & MP7.2) PoPs to GFP converter | ||
+ | |XbaI | ||
+ | |PstI | ||
+ | | | ||
+ | | | ||
+ | |[[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]] | ||
|} | |} | ||
== Strains (glycerol stock) == | == Strains (glycerol stock) == | ||
- | + | {| | |
- | + | |- style="background: #ccccff;" | |
- | + | ! colspan="5" style="background: #ccccff;" | Strains glycerol stock | |
+ | |- style="background: #ccccff; text-align: center;" | ||
+ | |width=5%| Number | ||
+ | |width=25%| Name | ||
+ | |width=55%| Description | ||
+ | |width=5%| Verified clone | ||
+ | |width=5%| Date | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S1 | ||
+ | |MG1655 | ||
+ | |Standard E.Coli strain | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S2 | ||
+ | |Acinetobacter ADP1 | ||
+ | |Strain used for TG coloration test with Red Nile | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S3 | ||
+ | |DH5alpha pKs::DGAT | ||
+ | |Enzyme catalysing the transormation of DAG into TG | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S4 | ||
+ | |<bbpart>BBa_pJ23100</bbpart> | ||
+ | |Strong constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23100 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S5 | ||
+ | |<bbpart>BBa_B0015</bbpart> | ||
+ | |Double terminator (clones 1 + 2) <partinfo>B0015 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | | | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S6 | ||
+ | |<bbpart>BBa_pJ23107</bbpart> | ||
+ | |Medium constitutive promoter in J61002 (clones 1 + 2) <partinfo>pJ23107 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 19| July 19]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S7 | ||
+ | |<bbpart>BBa_B0030</bbpart> | ||
+ | |Strong RBS (clones 1 + 2) <partinfo>B0030 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 19| July 19]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S8 | ||
+ | |<bbpart>BBa_E0422</bbpart> | ||
+ | |ECFP (RBS+LVA+Term) (clones 1 + 2) <partinfo>E0422 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 19| July 19]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S9 | ||
+ | |<bbpart>BBa_E0241</bbpart> | ||
+ | |PoPs to GFP converter (clones 1 + 2) <partinfo>E0241 DeepComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 19| July 19]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S10 | ||
+ | |<bbpart>BBa_E0840</bbpart> | ||
+ | |gfp tri-part; strong rbs (clones 1 + 2) <partinfo>E0840 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 19| July 19]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S11 | ||
+ | |<bbpart>BBa_J61047</bbpart> | ||
+ | |Cre ORF (clones 1 + 2) <partinfo>J61047 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 19| July 19]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S12 | ||
+ | |W121 | ||
+ | | | ||
+ | | | ||
+ | |05/07/07 | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S13 | ||
+ | |<bbpart>BBa_I0500</bbpart> | ||
+ | |pBAD AraC (clone 1+2) <partinfo>I0500 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 20| July 20]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S14 | ||
+ | |<bbpart>BBa_pJ23107</bbpart> | ||
+ | |Medium constitutive promoter in J61002 (clone 3+4) <partinfo>pJ23107 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 20| July 20]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S15 | ||
+ | |<bbpart>BBa_P1003</bbpart> | ||
+ | |Kanamycin resistance cassette (clone 1+2) <partinfo>P1003 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 20| July 20]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S16 | ||
+ | |<bbpart>J61002</bbpart>::<bbpart>I0500</bbpart> | ||
+ | |AraC pBad regulating mRFP expression in pJ61002 (L1) <partinfo>J61002 SpecifiedComponents</partinfo> | ||
+ | |L1.2, L1.3 | ||
+ | |[[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S17 | ||
+ | |<bbpart>pSB1A2</bbpart>::Lox66-DapA Coli | ||
+ | |Lox66-DapA Coli cloned as a biobrick (L2) | ||
+ | | | ||
+ | |[[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S18 | ||
+ | |<bbpart>pSB1A2</bbpart>::B0030 + J61047 | ||
+ | |RBS(B0030)-Cre ORF (L6) (clones 1+2) | ||
+ | | | ||
+ | |[[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S19 | ||
+ | |<bbpart>pSB1A2</bbpart>::<bbpart>BBa_J37026</bbpart> | ||
+ | |Lox66 (L9)(1 clone) <partinfo>J37026 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S20 | ||
+ | |<bbpart>pSB1A2</bbpart>::<bbpart>BBa_J37028</bbpart> | ||
+ | |Lox71 (L10)(1 clone) <partinfo>J37028 SpecifiedComponents</partinfo> | ||
+ | | | ||
+ | |[[Paris/July 27| July 27]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S21 | ||
+ | |<bbpart>pSB1A2</bbpart>::<bbpart>BBa_J37028</bbpart>+<bbpart>BBa_E0840</bbpart> | ||
+ | |Lox71-gfp tri-part (L12) | ||
+ | |L12.2 ok | ||
+ | |? | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S22 | ||
+ | |<bbpart>pSB1A2</bbpart>::Lox71-kmR-lox66 | ||
+ | |Lox71-kmR-lox66 (L25) | ||
+ | |sequence not good (thrown away) | ||
+ | |? | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S23 | ||
+ | |pTopo::P5 | ||
+ | |Lox66-DapAColi | ||
+ | | | ||
+ | |? | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S24 | ||
+ | |pTopo::P6 | ||
+ | |Lox66-DapASubtilis | ||
+ | | | ||
+ | |? | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S25 | ||
+ | |pTopo::P8 | ||
+ | |RBS-DapAColi | ||
+ | |MPT4.3, MPT4.5 | ||
+ | |? | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S26 | ||
+ | |pTopo::P9 | ||
+ | |RBS-DapASubtilis | ||
+ | | | ||
+ | |? | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S27 | ||
+ | |<bbpart>J61002</bbpart> | ||
+ | | | ||
+ | | | ||
+ | |[[Paris/August 14| August 14]] | ||
+ | |- style="background: #cccccc;" | ||
+ | | style="background: #ccffcc;" |S28 | ||
+ | |<bbpart>J61025</bbpart> | ||
+ | |FRT-Cm-FRT | ||
+ | | | ||
+ | |[[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]] | ||
+ | |} | ||
+ | |||
+ | |||
* p2C (CFP fluorescent) | * p2C (CFP fluorescent) | ||
* p2Y (YFP fluorescent) | * p2Y (YFP fluorescent) | ||
Line 224: | Line 1,656: | ||
** DCR 14.2 | ** DCR 14.2 | ||
** DCR 14.3 | ** DCR 14.3 | ||
- | |||
- | |||
- | |||
- | |||
- |
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