Table 1 Strains used in this study
E. coli DH10B | Cloning strain | Thermo scientific |
|---|---|---|
Strains | ||
T. kivui DSM 2030 | Acetogen, Topt = 66 °C | DSMZ |
TKV_0020 | Derivative of DSM 2030 | This work |
ASI_TKV_0001 | TKV_0020, SPF-B017 | This work |
ASI_TKV_0005 | TKV_0020, SPF-3037 (ΔpyrE) | This work |
ASI_TKV_0006 | TKV_0020, SPF-3038 (ΔpyrE) | This work |
ASI_TKV_0007 | TKV_0020, SPF-3039 (ΔpyrE) | This work |
ASI_TKV_0008 | TKV_0020, SPF-3040 (mini array) | This work |
ASI_TKV_0019 | DSM 2030, GGA with SPF-2070, 500 bp HAs (PCR SPF-3039 ASI229-230), SPF-B017 (ΔpyrE) | This work |
ASI_TKV_0020 | DSM 2030, DSM 2030, GGA with SPF-2070, 1000 bp HAs (PCR SPF-3039 ASI132-133), SPF-B017 (ΔpyrE) | This work |
ASI_TKV_0021 | DSM 2030, SPF-3078 (ΔpyrE) | This work |
ASI_TKV_0029 | DSM 2030, SPF-3047 (ΔrexA) | This work |
ASI_TKV_0031 | DSM 2030, SPF-3084 (ΔhrcA) | This work |
ASI_TKV_0032 | DSM 2030 SPF-3063 (ldh::pFAST) | This work |
ASI_TKV_0033 | DSM 2030 SPF-3064 (ldh::pFAST) | This work |
ASI_TKV_0041 | DSM 2030 SPF- B017 | This work |
ASI_TKV_0050 | DSM 2030 ΔhrcA, cured colony | This work |
ASI_TKV_0051 | DSM 2030, ldh::pFAST, cured colony | This work |
ASI_TKV_0052 | ASI_TKV_0051, SPF-3093 (ldh::pFAST, spacer 20) | This work |
ASI_TKV_0053 | ASI_TKV_0051, SPF-3094 (ldh::pFAST, spacer 15) | This work |
ASI_TKV_0054 | ASI_TKV_0051, SPF-3095 (ldh::pFAST, spacer 11) | This work |
RHO_TKV_0010 | TKV_0020, SPF-2015 (pThermoFAST_01) | [16] |
ALU_TKV_0009 | DSM 2030, SPF-4052 (PolCC629Y) | This work |
E. coli—Thermoanaerobacter shuttle plasmids | ||
SPF-B009 | pMU131 engineered for Bbs1 GGA cloning, Recipient Sites 1–4, removable amilCP | |
SPF-2021 | SPF-B009, active PAM and Spacer 1 (RH050-051) | This work |
SPF-2022 | SPF-B009, inactive PAM and Spacer 1 (RH052-053) | This work |
SPF-2023 | SPF-B009, active PAM and Spacer 2 (RH054-055) | This work |
SPF-2024 | SPF-B009, inactive PAM and Spacer 2 (RH056-057) | This work |
SPF-B017 | B6A-RI ori engineered for Bsa1 GGA cloning, recipient Sites A-D, colE1 origin of replication, thermoKanR | This work |
SPF-2061 | Synthetic mini array containing direct repeats, Esp3I recipient sites 2,3, colE1, ampR | This work |
SPF-2068 | SPF-2061, crRNA 1 (RH058-059) for pyrE | This work |
SPF-2069 | SPF-2061, crRNA 2 (RH060-061) for pyrE | This work |
SPF-2070 | SPF-2061, crRNA 3 (RH062-063) for pyrE | This work |
SPF-3037 | SPF-2068, 1000 bp HAs (ASI132-133), SPF-B017 | This work |
SPF-3038 | SPF-2069, 1000 bp HAs (ASI132-133), SPF-B017 | This work |
SPF-3039 | SPF-2070, 1000 bp HAs (ASI132-133), SPF-B017 | This work |
SPF-3040 | SPF-2061 (AD PCR, ASI168-169), SPF-B017 | This work |
SPF-3041 | SPF-2068 (AD PCR, ASI168-169), SPF-B017 | This work |
SPF-3042 | SPF-2069 (AD PCR, ASI168-169), SPF-B017 | This work |
SPF-3043 | SPF-2070 (AD PCR, ASI168-169), SPF-B017 | This work |
SPF-3077 | SPF-2070, 100 bp HAs (PCR SPF-3039 ASI231-232), SPF-B017 | This work |
SPF-3078 | SPF-2070, 500 bp HAs (PCR SPF-3039 ASI229-230), SPF-B017 | This work |
SPF-2094 | SPF-2061, crRNA hrcA (ASI145-146) | This work |
pMini2.0 | PCR cloning kit vector | NEB |
SPF-2157 | hrcA right HA (1000 bp, ASI151-152), pMini2.0 | This work |
SPF-2158 | hrcA left HA (1000 bp, ASI153-154), pMini2.0 | This work |
SPF-3084 | SPF-2094, SPF-2157, SPF-2158, SPF-B017 | This work |
SPF-2093 | SPF-2061, crRNA for rexA (ASI160-161) | This work |
SPF-3047 | SPF-2093, rexA HAs (1000 bp, ASI164-165, ASI166-167), SPF-B017 | This work |
SPF-2119 | SPF-2061, crRNA 1 for ldh (ASI195-196) | This work |
SPF-2120 | SPF-2061, crRNA 2 for ldh (ASI197-198) | This work |
SPF-2159 | ldh right HA (1000 bp, ASI199-200), pMini2.0 | This work |
SPF-2015 | SPF-B009, Expression of pFAST with Ppta, tKan | [16] |
SPF-2160 | ldh left HA (1000 bp, ASI201-202), pMini2.0 | This work |
SPF-3063 | SPF-2119, SPF-2159, PCR XY SPF-2015 (ASI203-204), SPF-2160, SPF-B017 | This work |
SPF-3064 | SPF-2120, SPF-2159, PCR XY SPF-2015 (ASI203-204), SPF-2160, SPF-B017 | This work |
SPF-3093 | crRNA (20 bp) for pFAST (ASI240-241), SPF-B017 | This work |
SPF-3094 | crRNA (15 bp) for pFAST (ASI242-243), SPF-B017 | This work |
SPF-3095 | crRNA (11 bp) for pFAST (ASI244-245), SPF-B017 | This work |
SPF-B030 | Same as SPF-B017 with the Pthl promoter (C. acetobutylicum) for thermoKanR | This work |
SPF-4051 | pTwist, AmpR, crRNA for polC, polC HA (C669Y), fusion sites AD | Synthesis |
SPF-4052 | SPF-4051, SPF-B030 | This work |