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Table 1 Plasmids and bacterial strains used in this study

From: Identification of two genes required for heptadecane production in a N2-fixing cyanobacterium Anabaena sp. strain PCC 7120

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 Chen et al. (2015), Xu et al. (2015)
 pZR824 Kmr; integration vector, annealed oligonucleotides ZR165/ZR166 ligated to BglII-SpeI digested pRL278 to produce pZR824 This study
 pZR932 Kmr/Apr; alr528384 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 alr528384 within pZR932 This study
 pZR934 Kmr; BamHI and AvrII digested alr528384 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 alr528384 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; alr528384 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
  1. Apr: ampicillin resistance; Spr: spectinomycin resistance; Nmr/Kmr: neomycin–kanamycin resistance; Cmr/Emr: chloramphenicol–erythromycin resistance; F2: two FLAG epitopes; MCS: multiple cloning sites; RBS: ribosome-binding site; P-: promoter; orf: open reading frame