Table 1.
List of organisms and plasmids used/generated in this work
| Organism namea | Description | Source |
|---|---|---|
| Escherichia coli DH5α | Transformation/cloning strain | Invitrogen |
| Synechocystis sp. PCC 6803 | Wild-type strain | Pasteur Culture Collection |
| SNnK | Synechocystis mutants with replacement of the neutral site Nn by a kanamycin resistance cassette | This work |
| SNnKS | Synechocystis mutants with replacement of the neutral site Nn by a kanamycin resistance/sucrose sensitivity cassette | This work |
| SNnK.gfp | Synechocystis mutants with replacement of the neutral site Nn by a kanamycin resistance cassette and the promoterless GFP encoding sequence | This work |
| SNnK.Cgfp | Synechocystis mutants with replacement of the neutral site Nn by a kanamycin resistance cassette and the GFP encoding sequence under the control of a constitutive promoter (PpsbA2*) | This work |
| Plasmid namea | Description | Source |
| pGEM®-T easy | T/A cloning vector | Promega |
| pK18mobsacB | Small mobilizable multi-purpose cloning vector derived from the E. coli plasmids pK18, conferring resistance to kanamycin (nptII), and sensitivity to sucrose (sacB) | National BioResource Project (NIG, Japan): E. coli |
| pSB1A2-E0240 | BioBrick™ vector containing the part BBa_E0240 (GFP generator) | Registry of Standard Biological Parts, MIT |
| pJ201-PpsbA2* | Synthesized minimal psbA2 promoter region, meeting the BioBrick RFC[10]standard specifications, cloned into the delivery vector pJ201 | DNA 2.0 Inc., this work |
| pBSK-FP300 | Synthetic interface containing a BioBrick-compatible multiple cloning site (MunI, NotI, XbaI, SpeI, NotI, PstI) insulated with the transcriptional terminators BBa_B1006 and BBa_B0062 (upstream), and BBa_B0053 and BBa_B020 (downstream) | Epoch Life Science Inc., this work |
| pSNn | pGEM-T easy-based plasmid series containing the BioBrick-compatible interface flanked by the two regions for double homologous recombination on neutral site n | This work |
| pSNnK | pSNn plasmid series with the kanamycin resistance cassette cloned upstream of the BioBrick-compatible interface | This work |
| pSNnKS | pSNn plasmid series with the kanamycin resistance/sucrose sensitivity cassette cloned upstream of the BioBrick-compatible interface | This work |
| pSNnK.gfp | pSNnK plasmid series with the GFP generator BioBrick BBa_E0240 cloned in the BioBrick-compatible interface | This work |
| pSNnK.Cgfp | pSNnK plasmid series with the GFP generator BioBrick BBa_E0240 under the control of a constitutive promoter (PpsbA2*), cloned in the BioBrick-compatible interface | This work |
an = 5, 8, 10, 15, 16.