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Synthetic Biology Research Centre

Synthetic Biology Research Centre

Anaerobe Knock-Out Vectors

Anaerobe Knock-Out Vectors

Description

Knock-out vectors are available for the generation of mutants in a range of anaerobes in a classic two-step process by allelic exchange using the counter selection marker codA (cytosine deaminase) or pyrE (orotate phosphoribosyltransferase)

[1] codA-based plasmids pMTL-SC7215, pMTL-SC7315 & pMTL-SC7515, previously exemplified in Clostridioides difficile and Clostridium acetobutylicum and based on the pBP1, pCB102 or pIM13 replicons, respectively.

[2] pyrE-based plasmids pMTL-ME3 & pMTL-GL3, carrying either the pIM13 replicon or none at all (suicide), respectively. Exemplified in C. acetotobutylicum and C. beijerinckii, respectively.

[3] pyrE-based plasmids pMTL-YN3 and pMTL-YN4, based on the pCB102 and pBP1 replicons and exemplified in different strains of Clostridioides difficile.

[4] pyrE-based suicide vector pMTL-JPB22 exemplified in Acetobacterium woodii.

Vectors supplied as a pair at a concentration of 50ng/µl DNA and are ready to be transformed into E. coli. Each pair costs 80 GBP plus postage at a rate dependent on destination - postage added at check-out.

 

Vectors are available for academic groups only, companies please contact [email protected]

https://plasmidvectors.com

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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Bacterial Strains Carrying Defined Mutants

Bacterial Strains Carrying Defined Mutants

Description

Defined mutants are available in various bacterial species (including, for instance) Clostridium, Cupriavidus, Geobacillus and Methanotrophs. Please identify beforehand exactly which mutants are required by contacting  [email protected] or [email protected] with your requirements.  

Mutants are sent as agar slabs or spore stocks. Postal charges are destination dependent and added at check-out.

Queries should be addressed to: [email protected] 

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering. 

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CRISPR-Cas9

CRISPR-Cas9

Description

Many CRISPR-Cas9 systems suffer from toxicity issues in both E. coli and the intended host as a consequence of overproduction of Cas9.  

Our Cas9 has a frameshift that results in a truncated Cas9 protein lacking 87 amino acids and is believed to act as a nickase.

The five vectors supplied are all based on the pCB102 Gram +ve replicon, rely on the Pthl promoter for truncated Cas9 expression and incorporate the listed component parts as (selectable marker - Gram negative replicon - sgRNA promoter):

pMTL431521 - (catP - ColE1/tra - PtcdB)
pMTL431511 - (catP - ColE1/tra - ParaE)
pMTL431211 - (catP - p15a/tra - ParaE)
pMTL432211 - (ermB - p15a/tra - ParaE)
pMTL431541 - (catP - ColE1/tra - PJ23119)

All vectors are supplied at a concentration of 50ng/µl DNA are ready to be transformed into E. coli. The kit costs 200 GBP plus postage at a rate dependent on destination - postage added at check-out.

Vectors are available for academic groups only, companies please contact [email protected]

https://plasmidvectors.com 

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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Donor Strains

Donor Strains

Description

E. coli strains which can be used as conjugative donors.

Plasmid DNA propagated in the strains varies in its adenosine and cytosine methylation status due to the presence or absence of functional  Dam and Dcm methyltransferases.  Each is Mob+ Tra+ and all but S17.1 carry the conjugative plasmid R702 (TcR, SmR, SuR, HgR).  

CA434 = HB101 (R702): F - mcrB mrr hsdS20(rB- mB-) recA13 leuB6 ara-14 proA2 lacY1 galK2 xyl-5 mtl-1 rpsL20 (SmR) glnV44 λ-)

sExpress = NEB Express (R702): fhuA2 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) endA1 Δ(mcrC-mrr)114::IS10

InterStellar = Stellar (R702): F – ara, Δ(lac-proAB) [Φ80d lacZΔM15], rpsL(str), thi, Δ(mrr-hsdRMS-mcrBC), ΔmcrA, dam, dcm

TopSex = Top10 (R702): mcrA, Δ(mrr-hsdRMS-mcrBC), Phi80lacZ(del)M15, ΔlacX74, deoR, recA1, araD139, Δ(ara-leu)7697, galU, galK, rpsL(SmR), endA1, nupG

S17.1:  pro, res− hsdR17 (rK− mK+) recA− with an integrated RP4-2-Tc::Mu-Km::Tn7, TpR - λpir.

Strains are sent as agar slabs. Postal charges are destination dependent and added at check-out.

Parental strains Express and Stellar are the property of New England Biolabs and CLONTECH, respectively.  

Queries should be addressed to: [email protected] 

https://plasmidvectors.com

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering. 

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Geobacillus CRISPR-Cas9

Geobacillus CRISPR-Cas9

Description

A universal CRISPR_Cas9 gene editing system for broad application in thermophilic Parageobacillus and other anaerobic Gram +ve thermophiles, comprising 2 vectors.

The widely used CRISPR/Cas9 nuclease of Streptococcus pyogenes is inactive at temperatures above 42 °C, making its use in thermophiles problematical. To circumvent this issue we have developed two Parageobacillus/E.coli shuttle vector CRISPR/Cas9 systems based on the well-characterized CRISPR/Cas9 systems of Streptococcus thermophiles, stCas91 and stCas93.

https://doi.org/10.1021/acssynbio.1c00138

Two vectors are included in the kit:-

pMTL575555 (stCas91)
pMTL675555 (stCas93)

Both vectors carry the ColEI and pUB110 origins of replication, a thermostable kanamycin resistance gene and a locus encoding the specific sgRNA.

All vectors are supplied at a concentration of 50ng/µl DNA are ready to be transformed into E. coli. The kit costs 200 GBP plus postage at a rate dependent on destination - postage added at check-out.

Vectors are available for academic groups only, companies please contact [email protected]

https://plasmidvectors.com 

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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pMTL60000 Modular Plasmid Vectors

pMTL60000 Modular Plasmid Vectors

Description

Modular shuttle plasmid vectors for use in Parageobacillus and other Gram-positive thermophiles.

Comprising a selection of Gram +ve and -ve replicons, antibiotic markers and various different application specific modules.

The 20 modules described are distributed as part of a kit comprising seven plasmids (pMTL61221, pMTL62112, pMTL63223, pMTL64224, pMTL65341, pMTL66331, and pMTL67331) from which the other possible combinations of modules can be easily constructed in one or a few cloning step(s) using AscI, FseI, PmeI and SbfI.

Plasmids are sent as lyophilised 50ng/µl DNA ready to be transformed into E. coli. and cost 280 GBP per Kit, plus postage at a rate dependent on destination - shipping added at checkout.

For further information on the vectors see https://plasmidvectors.com

Queries should be addressed to: [email protected]

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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pMTL70000 Modular Plasmid Vectors

pMTL70000 Modular Plasmid Vectors

Description

Modular shuttle plasmid vectors for use in Cupriavidus and other Gram-negative aerobes.

Comprising a selection of Cupriavidus and E. coli replicons, antibiotic markers and various different application specific modules.

KIT 1

The 18 modules originally described are distributed as part of a kit comprising six plasmids (pMTL70621, pMTL71302, pMTL72113, pMTL73414, pMTL74131 & pMTL75511) from which all other possible combinations of modules can be easily constructed in one or a few cloning step(s) using AscI, FseI, PmeI and SbfI.

For further information on the vectors see https://plasmidvectors.com 

Queries should be addressed to: [email protected] 

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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pMTL80000 Modular Plasmid Vectors

pMTL80000 Modular Plasmid Vectors

Description

Modular shuttle plasmid vectors for use in Clostridium bacteria and other Gram-positive anaerobes.

Comprising a selection of Gram +ve and -ve replicons, antibiotic markers and various different application specific modules.

KIT 1

The 18 modules originally described are distributed as part of a kit comprising five plasmids (pMTL80110, pMTL82254, pMTL83353, pMTL84422 & pMTL85141) from which the other 395 possible combinations of modules can be easily constructed in one or a few cloning step(s) using AscI, FseI, PmeI and SbfI.

KIT 2

Seven plasmids (pMTL89151, pMLT86151, pMTL87151, pMTL81151, pMTL88155, pMTL84131, pMLT82255) comprising five new Gram-positive replicons and the fluorescent marker FAST - for details, see https://plasmidvectors.com

Plasmids are sent as lyophilised 50ng/µl DNA ready to be transformed into E. coli. and cost 200 GBP per Kit, plus postage at a rate dependent on destination - shipping added at checkout.

For further information on the vectors see https://plasmidvectors.com 

Queries should be addressed to: [email protected] 

pMTL80000 modular plasmids at ClosTron http://clostron.com/pMTL80000.php 

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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RiboCas

RiboCas

Description

A universal CRISPR_Cas9 gene editing system for broad application in Clostridia and other anaerobic Gram +ve species, comprising 5 vectors.

In RiboCas, Cas9 has been placed under the control of a tightly regulated, theophylline responsive riboswitch. The system overcomes may of the issues relating to Cas9 toxicity in both the shuttle host and intended target.

https://doi.org/10.1021/acssynbio.9b00075 

Supplied as a kit, comprising five vectors, incorporating two different riboswitches controlling Cas9 (E & G), and a choice of promoter controlling the expression of the sgRNA, either ParaE (1) or the J23119 promoter (2).  Four of the plasmids are based on the ColE1 replicon, while the fifth uses the low copy number, p15a replication region.

pRECas1
pRECas2
pRECas1-p15a
pRGCas1
pRGCas2

All vectors are supplied at a concentration of 50ng/µl DNA ready to be transformed into E. coli. The kit costs 200 GBP plus postage at a rate dependent on destination – postage added at check-out.

Vectors are available for academic groups only, companies please contact [email protected] 

External URL: https://plasmidvectors.com 

Please note: It is the responsibility of the buyer to make sure that prior to purchase that the required goods are accepted into the destination country and that any required documentation (including import permits) are in place. Refunds cannot be processed for any orders returned to us due to customs issues. Please contact [email protected] for any questions relating to ordering.

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