Paris/Freezer
From 2007.igem.org
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| DGAT deltaPst1 | | DGAT deltaPst1 | ||
|1504 | |1504 | ||
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| + | |[[Paris/August_2| August 2]] | ||
| + | |- style="background: #cccccc;" | ||
| + | | style="background: #ccffcc;" |P15 | ||
| + | | [pTet>>lox-GFP-loxmRFP] L59.6 | ||
| + | | O47 | ||
| + | | O48 | ||
| + | | PacI [pTet>>lox-GFP-loxmRFP] XmaI | ||
| + | |2kb | ||
| + | | | ||
| + | | | ||
| + | | | ||
| + | |[[Paris/August_2| August 2]] | ||
| + | |- style="background: #cccccc;" | ||
| + | | style="background: #ccffcc;" |P15 | ||
| + | | [pTet>>lox-GFP-loxmRFP] L59.7 | ||
| + | | O47 | ||
| + | | O48 | ||
| + | | PacI [pTet>>lox-GFP-loxmRFP] XmaI | ||
| + | |2kb | ||
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Revision as of 17:10, 15 October 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 | |||
| P15 | [pTet>>lox-GFP-loxmRFP] L59.6 | O47 | O48 | PacI [pTet>>lox-GFP-loxmRFP] XmaI | 2kb | August 2 | |||
| P15 | [pTet>>lox-GFP-loxmRFP] L59.7 | O47 | O48 | PacI [pTet>>lox-GFP-loxmRFP] XmaI | 2kb | August 2 | |||
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 | |||
| L10 | lox71 in pSB1A2 | |||
| 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
