Table A2.
Plasmids used in this study
| Plasmid number | Description | Primers used | Enzymes used | Source/reference |
|---|---|---|---|---|
| pTA131 | pyrE2 marked integrative vector for knock‐outs in Haloferax volcanii (Ampr) | – | – | Allers et al., 2004 |
| pTA298 | Vector containing trpA marker under a ferredoxin promoter between two BamHI sites (Ampr) | – | – | Lestini et al., 2010 |
| pTA356 | hdrB marked shuttle vector with pHV1 replication origin (Ampr) | – | – | Norais et al., 2007 |
| pTA409 | pyrE2 and hdrB marked shuttle vector with pHV1 replication origin (Ampr) | – | – | Delmas et al., 2009 |
| pTA1228 | pyrE2 and hdrB marked expression vector with pHV2 replication origin and tryptophanase promoter p.tnaA, for tryptophan‐inducible gene expression of N‐terminal 6x His‐tagged protein (Ampr) | – | – | Brendel et al., 2014 |
| pTA1369 | pyrE2 marked vector containing a tryptophan promotor (p.tnaA)‐multiple cloning site‐construct with a hdrB marker under a ferredoxin promotor between two BglII sites; used to generate a tryptophan‐inducible allele of gene (Ampr) | – | – | This study; derived from pTA131 and pTA1228 (see Figure A11) |
| pTA1392 | pyrE2 and hdrB marked expression vector pTA1228 with insertion of Strep II tag, for tryptophan‐inducible gene expression of N‐terminal 6x His‐tagged and/or C‐terminal StrepII‐tagged protein (Ampr) | – | – | Gamble‐Milner, 2016 |
| pTA1451 | pyrE2 marked vector containing a tryptophan promotor with reduced leak activity (p.tnaM3)‐multiple cloning site‐construct with a hdrB marker under a ferredoxin promotor between two BglII sites; used to generate a tryptophan‐inducible allele of gene (Ampr) | – | ClaI; BclI; NdeI | This study; p.tnaM3 promoter from pNPM‐tnaM3‐HfxMCM (S. MacNeill, unpublished)(see Figure A11) |
| pTA1992 | pyrE2 and hdrB marked expression vector with pHV2 replication origin and synthetic promoter (p.syn), for constitutive gene expression of N‐terminal 6x His‐tagged and/or C‐terminal StrepII‐tagged protein (Ampr) | p.synF; p.synR2 | BstBI; NdeI | This study |
| p7XNH3 | Expression vector for E. coli using T7‐promotor with N‐terminal 10x His‐Tag (FX‐Cloning system) (Kanar) | – | – | Geertsma & Dutzler, 2011 |
| pSVA3930 | pTA131‐based deletion construct for dacZ containing ~500 bp upstream and downstream flanking regions ligated via BamHI site (Ampr) | 8001; 8002; 8003; 8004 | KpnI; BamHI; XbaI | This study |
| pSVA5408 | p7XNH3‐based expression vector for heterologous expression of dacZ with N‐terminal 10x His‐Tag in Escherichia coli (Kanar) | 7644; 7645 | SapI | This study |
| pSVA5411 | pTA1369‐based vector containing a tryptophan‐inducible (p.tnaA) allele of dacZ with a hdrB marker under a ferredoxin promotor between two BglII sites (Ampr) | 7654; 7662 | NdeI; EcoRI | This study |
| pSVA5414 | pSVA3930‐based integrative vector with tryptophan‐inducible (p.tnaA) allele of dacZ with a hdrB marker under a ferredoxin promotor inserted between upstream and downstream flanking regions at BamHI site (Ampr) | – | BglII; BamHI | This study |
| pSVA5417 | pTA1992‐based expression vector for homologous expression of dacZ with N‐terminal 6x His‐Tag in H. volcanii (Ampr) | 7669; 7633 | PciI; NcoI; EcoRI | This study |
| pSVA5418 | pTA131‐based deletion construct for predicted Operon dacZ+mscS2 containing ~500 bp upstream and downstream flanking regions ligated via BamHI site (Ampr) | 8001; 7672 | KpnI; BamHI; XbaI | This study |
| pSVA5419 | pTA1451‐based vector containing a tryptophan‐inducible (p.tnaM3) allele of dacZ with a hdrB marker under a ferredoxin promotor between two BglII sites (Ampr) | – | NdeI; XhoI | This study |
| pSVA5420 | pSVA3930‐based integrative vector marked with trpA under a ferredoxin promoter ligated at BamHI site (Ampr) | – | BamHI | This study |
| pSVA5421 | pTA356‐based episomal vector containing dacZ with ~1,000 bp upstream and downstream flanking regions (Ampr) | 7680; 7681 | KpnI; BamHI | This study |
| pSVA5422 | pTA409‐based episomal vector containing dacZ with ~1,000 bp upstream and downstream flanking regions (Ampr) | 7680; 7681 | KpnI; BamHI | This study |
| pSVA5427 | pTA131‐based deletion construct for mscS2 containing ~500 bp upstream and downstream flanking regions ligated via BamHI site (Ampr) | 7690; 7693 | KpnI; BamHI; XbaI | This study |
| pSVA5433 | pSVA5417‐based expression vector for homologous expression of dacZ(D194A) with N‐terminal 6x His‐Tag in H. volcanii (Ampr) | 9804; 9805 | – | This study |