Paris/August 20
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Ligations
| Ligations | ||||||
|---|---|---|---|---|---|---|
| Number | Insert | Insert Volume (µL) | Vector | Vector Volume (µL) | Comments | Number of colonies |
| C1 | MP3.1 digested with EcoRI (D0) | 2 | control of ligation efficiency | |||
| C3 | D34 (lox66 BV) | 4µl | control of D34 digestion | |||
| C5 | D50 (BBa_J61002 BV) | 1.5µl | control of D50 digestion | |||
| C6 | D41 (araC/pBad-J61002 plasmid SP dig.) | 1µl | control of D41 digestion | |||
| C7 | D22 (pSB1A2 open vector EP) | 4µl | control of D22 digestion | |||
| C8 | D47 (pSB1A2 XP dig.) | 4µl | control of D47 digestion | |||
| C9 | D48 (pSB1A2 ES dig.) | 2µl | control of D48 digestion | |||
| L31 | D44 (B0015 terminator BI) | 4 | D35 (lox71 BV) | 3 | Insertion of B0015 terminator behind lox71 | +++ |
| L32 | D45 (B0015 terminator FI) | 4 | D42 (Lox66 FV) | 3 | Insertion of B0015 terminator in front of lox66 | +++ |
| L33 | D33.1 (RBS(B0030)CreORF BI) | 5 | D50 (BBa_J61002 BV) | 1.5 | pTet>>RBS-Cre | RAS |
| L34 | D49 (lox71-gfptripart BI) | 5 | D50 (BBa_J61002 BV) | 1.5 | pTet>>lox71-gfptripart | ++ |
| L35 | D49 (lox71-gfptripart BI) | 5 | D41 (araC/pBad-J61002 plasmid SP dig.) | 1 | araC/pBad>>lox71-gfptripart | +++ |
| L36 | D13.4 (mRFP dig. SP) | 6 | D34 (lox66 BV) | 4 | lox66-mRFP | +++ |
| L37 | D52 (RBS-dapAcoli BI) | 6 | D34 (lox66 BV) | 4 | lox66-RBS-dapAcoli | +++ |
| L38 | D54 (RBS-dapAsubtilis BI) | 13 | D34 (lox66 BV) | 4 | lox66-RBS-dapAsubtilis | RAS |
| L39 | D51 (RBS-dapAcoli BBinsert) | 4 | D22 (pSB1A2 open vector EP) | 4µl | RBS-dapAcoli cloned in pSB1A2 | +++ |
| L40 | D53 (RBS-dapAsubtilis BBinsert) | 13 ( | D22 (pSB1A2 open vector EP) | 4µl | RBS-dapAsubtilis cloned in pSB1A2 | + |
| L41 | D27' (lox71-ftsZ bw-insert) | 12 | D47 (pSB1A2 XP dig.) | 4µl | lox71-ftsZ cloned in pSB1A2 | + |
| L42 | D32 (lox71-ftsAftsZ bw-insert) | 6 | D47 (pSB1A2 XP dig.) | 4µl | lox71-ftsA-Z cloned in pSB1A2 | + |
| L43 | D55 (RBS-dapAsubtilis FI) | 15 | D48 (pSB1A2 ES dig.) | 2µl | RBS-dapAsubtilis cloned in pSB1A2 | + |
5 points with :
- L31
- L32
- L34
- L35
- L36
- L37
- L39
- L40 (2 points only)
- L41 (2 points only)
- L42
- L43
Sequencing
The following sequencing reactions have been performed
TL_MPT5.1+O41 TL_MPT5.2+O41 TL_L26.1+O18 TL_L26.1+O19 TL_L26.2+O18 TL_L26.2+O19 TL_L27.2+O18 TL_L27.2+O19 TL_L27.3+O18 TL_L27.3+O19 TL_L27.4+O18 TL_L27.4+O19 TL_L27.5+O18 TL_L27.5+O19 TL_L28.2+O18 TL_L28.2+O19 TL_L28.3+O18 TL_L28.3+O19 TL_L28.8+O18 TL_L28.8+O19 TL_L29.1+O18 TL_L29.1+O19 TL_L29.2+O18 TL_L29.2+O19 TL_L30A1+O18 TL_L30A1+O19 TL_L30B1+O18 TL_L30B1+O19 TL_L27.2+O24 TL_L27.3+O24 TL_L27.4+O24 TL_L27.5+O24 TL_L28.2+O24 TL_L28.3+O24 TL_L28.8+O24 TL_L28.2+O22 TL_L28.3+O22 TL_L28.8+O22 TL_L28.2+O23 TL_L28.3+O23 TL_L28.8+O23
