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Table 1 Strains and plasmids used in this study

From: Mutant generation by allelic exchange and genome resequencing of the biobutanol organism Clostridium acetobutylicum ATCC 824

Strain/plasmid

Relevant features

Reference/source

Strains

 E. coli Top10

F- mcrA Δ(mrr-hsdRMS-mcrBC) Φ80lacZΔM15 ΔlacX74 deoR recA1 araD139 Δ(ara-leu)7697 galU galK rpsL (StrR) endA1 nupG

Invitrogen

 C. acetobutylicum

ATCC 824 (COSMIC)

CRG1268 strain used in the ERANET (http://www.sysmo.net) SysMO project COSMIC. Originally from G. Bennet, RICE University, USA

H Bahl University of Rostock

 C. acetobutylicum

ATCC 824

CRG3286, resourced from the ATCC in November 2011

ATCC

 C. acetobutylicum

CRG1545

ΔpyrE mutant of CRG1268 made using the ACE vector pMTL-JH12

[24]

 C. acetobutylicum

CRG3899

ΔpyrE mutant of CRG3286 equivalent to CRG1268

This study

 C. acetobutylicum

CRG3520

Δspo0A of CRG1545 (ΔpyrE) made using pMTL-ME3::spo0A

This study

 C. acetobutylicum

CRG4889

Δspo0A mutant of CRG1545 generated from CRG3520 by restoration of the pyrE allele to wild-type using pMTL-ME6

This study

 C. acetobutylicum

CRG4890

CRG3520 (Δspo0A) carrying a functional copy of spo0A and its own promoter inserted into the genome using the ACE vector pMTL-ME6X::spo0A concomitant with restoration of the pyrE allele

This study

 C. acetobutylicum CRG4891

CRG3520 (Δspo0A) carrying a functional copy of spo0A under the control of the fdx promoter inserted into the genome using the ACE vector pMTL-ME6X::spo0A concomitant with restoration of pyrE

This study

 C. acetobutylicum

CRG4893

In-frame deletion mutant of amyP (CaP0168) made using pMTL-ME3::amyP (pyrE vector with amyP KO cassette)

This study

 C. acetobutylicum

CRG4894

restoration of the pyrE allele to wild-type using pMTL-ME6

This study

 C. acetobutylicum

CRG4896

CRG4893 (ΔamyP) carrying a functional copy of amyP and its own promoter inserted into the genome using the ACE vector pMTL-ME6C::amyP concomitant with restoration of the pyrE allele

This study

 C. acetobutylicum

CRG4897

CRG4894 (ΔamyP) carrying a functional copy of amyP under the control of the fdx promoter inserted into the genome using the ACE vector pMTL-ME6X::amyP concomitant with restoration of pyrE allele

This study

 C. acetobutylicum

CRG4883

In-frame deletion mutant of glgA (Cac2239) made using pMTL-ME3::glgA (pyrE vector with glgA KO cassette)

This study

 C. acetobutylicum

CRG4898

ΔglgA mutant of CRG4883 generated from CRG1545 by restoration of the pyrE allele to wild-type using pMTL-ME6

This study

 C. acetobutylicum

CRG4901

CRG4883 (ΔglgA) carrying a functional copy of glgA and its own promoter inserted into the genome using the ACE vector pMTL-ME6C::glgA concomitant with restoration of the pyrE allele

This study

 C. acetobutylicum

CRG4902

CRG4883 (ΔglgA) carrying a functional copy of glgA under the control of the fdx promoter inserted into the genome using the ACE vector pMTL-ME6X::glgA concomitant with restoration of pyrE allele

This study

 C. acetobutylicum CRG2404

In-frame deletion mutant of spo0A gene made using pMTL-SC7515::spo0A

This study

 C. acetobutylicum CRG2403

In-frame deletion mutant of the type II Restriction system Cac1502 made using pMTL-SC7515::cac824I

This study

Plasmids

  

 pAN-2

E.coli vector (Ï•3T I methyltransferase gene, tet, p15a)

CRG685 [13]

 pMTL85141

E.coli- Clostridium Shuttle vector (pIM13 catP ColEI)

CRG1000 [31]

 pMTL-ME2

Clostridium KO vector (pCB102 catP ColE1 pyrE)

CRG1668 [25]

 pMTL-ME3

Clostridium KO vector (pIM13 catP ColE1 pyrE)

CRG1671This study

 pMTLSC7315

Clostridium KO vector (pCB102 catP ColE1 traJ codA)

CRG1424 [21]

 pMTLSC7515

Clostridium KO vector (pIM13 catP ColE1 traJ codA)

CRG1303 This study

 pMTL-ME6

C. acetobutylicum ACE correction vector (pIM13 catP ColE1 `pyrE)

CRG1861 This study

 pMTL-ME6C

C. acetobutylicum ACE complementation vector (pIM13 catP ColE1 pyrE lacZ’ MCS)

CRG2483 [25]

 pMTL-ME6X

C. acetobutylicum ACE expression vector (pIM13 catP ColE1 `pyrE P fdx, lacZ’ MCS)

CRG2482 [25]

 pMTL-ME3::spo0A

pMTL-ME3 + spo0A KO cassette (pIM13 catP ColE1 pyrE)

CRG1701 This study

 pMTL-ME3::amyP

pMTL-ME3 + amyP KO cassette (pIM13 catP ColE1 pyrE)

CRG3727 This study

 pMTL-ME3::glgA

pMTL-ME3 + glgA KO cassette (pIM13 catP ColE1 pyrE)

CRG4881 This study

 pMTL-SC7515::spo0A

pMTLSC7515 + spo0A KO cassette (pIM13 catP ColE1 codA)

CRG1672 This study

 pMTL-SC7515::cac824I

pMTLSC7515 + cac824I KO cassette (pIM13 catP ColE1 codA)

CRG2293 This study

 pMTL-ME6C::spo0A

ACE complementation vector for spo0A

CRG3739 This study

 pMTL-ME6X::spo0A

ACE overexpression vector for spo0A

CRG3738 This study

 pMTL-ME6C::amyP

ACE complementation vector for amyP

CRG3723 This study

 pMTL-ME6X::amyP

ACE overexpression vector for amyP

CRG3722 This study

 pMTL-ME6C::glgA

ACE complementation vector for glgA

CRG4900 This study

 pMTL-ME6X::glgA

ACE overexpression vector for glgA

CRG4880 This study