Paris/Freezer
From 2007.igem.org
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| - | | 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) | ||
Revision as of 12:30, 26 July 2007
Contents |
PCR products
| PCR products | |||||
|---|---|---|---|---|---|
| Number | Name | Length | Comments | Product Left ? | Date |
| P1 | Lox71-FtsA-FtsZ-1 | 1537 | Nothing left | ||
| P2 | FtsZ-2 | 1088 | Nothing left | ||
| P3 | DGAT-1 | ||||
| P4 | DGAT-2 | ||||
| P5 | Lox66-DapAColi | 985 | |||
| P6 | Lox66-DapASubtilis | 979 | 17/07/07 | ||
| P7 | Lox71-FtsZ | 1260 | EcoRI site in FtsZ not mutated | 17/07/07 | |
| P8 | RBS-DapAColi | 952 | B0030-DapAColi | No | 24/07/07 |
| P9 | RBS-DapASubtilis | 946 | B0030-DapASubtilis | No | 24/07/07 |
| P10 | Lox71-FtsA-FtsZ | 2580 | EcoRI site in FtsZ not mutated | 24/07/07 | |
Plasmids: MiniPrep Products
| Plasmids: MiniPrep Products | ||||
|---|---|---|---|---|
| Number | Name | Description | Product finished ? | Date |
| MP1 | pJ23100 in J61002 | Strong constitutive promoter in J61002 (clones 1 + 2) | 15/07/07 | |
| MP2 | pJ23107 in J61002 | Medium constitutive promoter in J61002 (clones 1 + 2) | MP2.2 | 15/07/07 |
| MP3 | B0015 in pSB1AK3 | Double terminator (clones 1 + 2) | 16/07/07 | |
| MP4 | I0500 in pSB2K3 | AraC pBAD (clones 1 + 2) | 16/07/07 | |
| MP5 | BBa_B0030 in pSB1A2 | Strong RBS (clones 1 + 2) | 18/07/07 | |
| MP6 | BBa_E0422 in pSB1A2 | ECFP (RBS+LVA+Term) (clones 1 + 2) | 18/07/07 | |
| MP6' | BBa_E0422 in pSB1A2 | ECFP (RBS+LVA+Term) (clones 1 + 2) | 24/07/07 | |
| MP7 | BBa_E0241 in pSB1A2 | PoPs to GFP converter (clones 1 + 2) | 18/07/07 | |
| MP7' | BBa_E0241 in pSB1A2 | PoPs to GFP converter (clones 1) | 24/07/07 | |
| MP8 | BBa_E0840 in pSB1A2 | gfp tri-part; strong rbs (clones 1 + 2) | 18/07/07 | |
| MP9 | BBa_J61047 in pSB1A2 | Cre ORF (clones 1 + 2) | 18/07/07 | |
| MP9' | BBa_J61047 in pSB1A2 | Cre ORF (clones 1 + 2) | 24/07/07 | |
| MP10 | BBa_I0500 in pSB2K3 | AraC pBAD (clones 1 + 2) | 20/07/07 | |
| MP11 | BBa_pJ23107 in p61002 | Medium constitutive promoter in J61002 (clones 3 + 4) | MP11.3 | 20/07/07 |
| MP11' | BBa_pJ23107 in p61002 | Medium constitutive promoter in J61002 (clones 3) | 23/07/07 | |
Digestion Products
PURPLE = Vector
GREEN = Insert
| Digestion Products | ||||||||
|---|---|---|---|---|---|---|---|---|
| Number | Product Name | Matrix Name | Enzyme 1 | Enzyme 2 | Size | Description | Product Left | Date |
| D1 | pJ23100 | pJ23100 | Spe1 | Pst1 | 2096 | Vector with the J23100 promoter ready for insertion of a RBS/CDS insert | 17/07/07 | |
| D2 | mRFP | pJ23100 | Spe1 | Pst1 | 887 | 17/07/07 | ||
| D3 | pJ23107 | pJ23107 | Spe1 | Pst1 | 2096 | Vector with the J23107 promoter ready for insertion of a RBS/CDS insert | 17/07/07 | |
| D4 | mRFP | pJ23107 | Spe1 | Pst1 | 887 | 17/07/07 | ||
| 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. | 17/07/07 | |
| 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. | 17/07/07 | |
| D7 | Lox66 DapA E.coli | Lox66 DapA E.coli (PCR product) | EcoR1 | Pst1 | 977 | 17/07/07 | ||
| D8 | Lox66 DapA E.coli | Lox66 DapA E.coli (PCR product) | Xba1 | Pst1 | 962 | 17/07/07 | ||
| D9 | pJ23100 ES vector | pJ23100 (MP1.1 & MP1.2) | EcoI | SpeI | 2925 | Vector with mRFP RBS/CDS ready for insertion promoter (fw-insert) | 21/07/07 | |
| D10 | pJ23100 promoter fw-insert | pJ23100 (MP1.1 & MP1.2) | EcoI | SpeI | 58 | J23100 promoter ready for use as a fw-insert | 21/07/07 | |
| 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 | 21/07/07 | |
| 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 | 21/07/07 | |
| 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 | 21/07/07 | |
| D14 | Cre ORF | J61047 MP9.1 & MP9.2 | XbaI | PstI | 1063 | Cre ORF extracted for usage as a bw-insert | 21/07/07 | |
| D15 | B0030 bw-vector | B0030 MP5.1 & MP5.2 | SpeI | PstI | 2076 | bacward vector for cloning a CDS bw-insert after the B0030 RBS | 21/07/07 | |
| D18 | BB dig. lox66DapAE.coli | lox66-DapAE.coli P5 PCR product | EcoRI | PstI | ~960 | PCR product digested for cloning as a BioBrick | Ø | 21/07/07 |
| D19 | BB dig. lox66DapAsubtilis | lox66-DapAsubtilis P6 PCR product | EcoRI | PstI | ~960 | PCR product digested for cloning as a BioBrick | Ø | 21/07/07 |
| 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 | 21/07/07 |
| D21 | bw-insert lox66-DapAsubtilis | lox66-DapAsubtilis P6 PCR product | XbaI | PstI | ~960 | PCR product digested for usage as a backward insert | Ø | 21/07/07 |
| D22 | pSB1A2 open vector | BBa_J61047 (MP9.1 & MP9.2) | EcoI | PstI | Open pSB1A2 vector for BioBrick EcoRI/PstI insertio | D22.1 Ø | 23/07/07 | |
| 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 | 23/07/07 | |
| D24 | eCFP bw-insert | BBa_E0422 (MP6.1 & MP6.2) | XbaI | PstI | eCFP (RBS-OFR_LVA-T) ready for use as a backward insert | 23/07/07 | ||
| D25 | PoPs to GFP converter | BBa_E0421 (MP7.1 & MP7.2) | XbaI | PstI | PoPs to GFP converter ready for use as a backward insert | 23/07/07 | ||
| D26 | gfp-tripart | BBa_E0840 (MP8.1 & MP8.2) | XbaI | PstI | gfp-tri part (strongRBS-ORF-T) ready for use as a backward insert | 23/07/07 | ||
| D27 | lox71-ftsZ bw-insert | lox71-ftsZ PCR product P7 | XbaI | PstI | lox71-ftsZ(unmutated) ready for use as a backward insert | Ø | 23/07/07 | |
Unexpected digestion results
| Unexpected results | ||||||
|---|---|---|---|---|---|---|
| Number | Matrix Name | Enzyme 1 | Enzyme 2 | Expected Size | Description | Date |
| D16 | pSB1AK3::B0015 | 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 | 21/07/07 |
| D17 | pSB1A2::E0422 (MP6.1 & MP6.2) | EcoRI | PstI | ~2040 + ~920 | 3 bands | 21/07/07 |
Strains (glycerol stock)
| Strains glycerol stock | ||||
|---|---|---|---|---|
| Number | Name | Description | 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) | 19/07/07 | |
| S7 | BBa_B0030 | Strong RBS (clones 1 + 2) | 19/07/07 | |
| S8 | BBa_E0422 | ECFP (RBS+LVA+Term) (clones 1 + 2) | 19/07/07 | |
| S9 | BBa_E0241 | PoPs to GFP converter (clones 1 + 2) | 19/07/07 | |
| S10 | BBa_E0840 | gfp tri-part; strong rbs (clones 1 + 2) | 19/07/07 | |
| S11 | BBa_J61047 | Cre ORF (clones 1 + 2) | 19/07/07 | |
| S12 | W121 | 05/07/07 | ||
| S13 | BBa_I0500 | pBAD AraC (clone 1+2) | 20/07/07 | |
| S14 | BBa_pJ23107 | Medium constitutive promoter in J61002 (clone 3+4) | 20/07/07 | |
- 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
