Table 1 Plasmids and bacterial strains used in this study
Plasmid or strain | Relevant characteristic(s) | Source or references |
|---|---|---|
Plasmids | ||
 pAM1956 | Promoter-less GFPmut2 cloning vector | Yoon and Golden (1998) |
 pRL271 | Cmr/Emr. Integration vector | Cai and Wolk (1990) |
 pRL278 | Kmr/Nmr, integration vector | Cai and Wolk (1990) |
 pRL443 | Apr; conjugal plasmid | Elhai et al. (1997) |
 pRL623 | Cmr; helper plasmid | Elhai et al. (1997) |
 pZR606 | Kmr/Spr, integration vector | Chen et al. (2015) |
 pZR618 | Apr; T7-MCS-F2-H6 | Chen et al. (2012) |
 pZR666 | Kmr/Spr, MCS-gfp-MCS cassette, annealed oligonucleotides LK2406/LK2407 ligated to AflII digested pZR606 to produce pZR666 | This study |
 pZR670 | Cmr/Emr; expression vector for Anabaena | |
 pZR824 | Kmr; integration vector, annealed oligonucleotides ZR165/ZR166 ligated to BglII-SpeI digested pRL278 to produce pZR824 | This study |
 pZR932 | Kmr/Apr; alr5283–84 ORF amplified by PCR with primers ZR241 and 242 from Anabaena 7120 chromosomal DNA and cloned into pCR2.1-TOPO vector | This study |
 pZR933 | Kmr/Apr; site-directed mutagenesis using primers ZR243 and 244 to introduce NotI site into alr5283–84 within pZR932 | This study |
 pZR934 | Kmr; BamHI and AvrII digested alr5283–84 ORF from pZR933 ligated to BglII and SpeI digested pZR824 | This study |
 pZR935 | Kmr/Spr; NotI and XbaI promotor-less GFP-Spr cassette from pZR666 ligated to NotI and NheI digested pZR934 to disrupt alr5283–84 | This study |
 pZR2222 | Kmr/Apr; Primers ZR1584 and ZR1585 PCR amplified 1921 bp Cmr/Emr cassette (NsiI/NdeI-NheI/BamHI-Emr/Cmr cassette-EcoRV/BglII/XhoI/XmaI/SmaI) from pRL271 ligated to pCR2.1-TOPO to produce pZR2222 | This study |
 pZR2223 | Cmr/Emr; expression vector, BamHI/XmaI cut out 1.9 kb Cmr/Emr cassette from pZR2222 and ligated to BglII/SgrAI-digested pAM1956 | This study |
 pZR2238 | Kmr/Apr; AatII/SalI-P-alr5283-P-alr5284-KpnI/XmaI PCR amplified by ZR1602 and 1603 from A. 7120 ligated to pCR2.1-TOPO | This study |
 pZR2239 | Cmr/Emr; SalI-P-alr5283-P-alr5284-XmaI from pZR2238 ligated to SalI/XmaI cut pZR2223 | This study |
 pZR2242 | Kmr/Apr; NsiI/NdeI-alr5283 orf-P-alr5284 orf-BamHI PCR amplified from A. 7120 ligated to pCR2.1-TOPO | This study |
 pZR2243 | Cmr/Emr; NsiI-alr5283 orf-P-alr5284 orf-BamHI from pZR2242 ligated to NsiI-BamHI cut pZR670 | This study |
 pZR2244 | Kmr/Apr; NsiI/NdeI-alr5283 orf PCR amplified by ZR1606 and 1608 from A. 7120; RBS-alr5284 orf-BamHI PCR amplified by ZR1609 and 1607 from A. 7120; PCR overlap of NsiI/NdeI-alr5283 orf and RBS-alr5284 orf-BamHI with primers ZR1606 and 1607; PCR overlap product ligated to pCR2.1-TOPO | This study |
 pZR2248 | Cmr/Emr; NsiI-alr5283 orf-RBS-alr5284 orf-BamHI from pZR2244 ligated to NsiI-BamHI cut pZR670 | This study |
Bacterial strains | ||
 TOP10 | E. coli cloning host | Invitrogen |
 NEB10β | E. coli cloning host | New England Biolabs |
 WT7120 | Anabaena sp. PCC 7120 wild-type strain | This study |
 DR935 | Spr; alr5283–84 double knockout mutant | This study |
 DR935(pZR2239) | Spr, Emr; DR935 containing pZR2239 for complementation study | This study |
 DR935(pZR2248) | Spr, Emr; DR935 containing pZR2248 for complementation study | This study |
 DR935(pZR2243) | Spr, Emr; DR935 containing pZR2243 for complementation study | This study |