| 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 BI
| ++
| L38
| D54 (RBS-dapAsubtilis BI)
| 13
| D34 (lox66 BV)
| 4
| lox66-RBS-dapAsubtilis BI
| 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
|
