Paris/Freezer

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Contents

PCR products

PCR products
Number Name Length Comments Product Left ? Date
P1 Lox71-FtsA-FtsZ-1 1537 Ø
P2 FtsZ-2 1088 Ø
P3 DGAT-1
P4 DGAT-2
P5 Lox66-DapAColi 985 Ø 17/07/07
P5' Lox66-DapAColi 985 27/07/07
P6 Lox66-DapASubtilis 979 Ø 17/07/07
P6' Lox66-DapASubtilis 979 27/07/07
P7 Lox71-FtsZ 1260 EcoRI site in FtsZ not mutated 17/07/07
P8 RBS-DapAColi 952 B0030-DapAColi Ø 24/07/07
P8' RBS-DapAColi 952 B0030-DapAColi 27/07/07
P9 RBS-DapASubtilis 946 B0030-DapASubtilis Ø 24/07/07
P9' RBS-DapASubtilis 946 B0030-DapASubtilis 27/07/07
P10 Lox71-FtsA-FtsZ 2580 EcoRI site in FtsZ not mutated 24/07/07
P11 Lox71-KmR-Lox66 1084 27/07/07

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 Ø D14.1 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 Ø D22.1 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 Ø D23.1 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 (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 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
S15 BBa_P1003 Kanamycin resistance cassette (clone 1+2) 20/07/07
S16 J61002::I0500 AraC pBad regulating mRFP expression in pJ61002 (L1) 27/07/07
S17 pSB1A2::Lox66-DapA Coli Lox66-DapA Coli cloned as a biobrick (L2) 27/07/07
S18 pSB1A2::B0030 + J61047 RBS(B0030)-Cre ORF (L6) 27/07/07
S19 pSB1A2::BBa_J37026 Lox66 (L9) 27/07/07
S20 pSB1A2::BBa_J37028 Lox71 (L10) 27/07/07


  • 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