Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004414 | pMTL84151 | In stock, 1 week for quality controls |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
The pMTL84151 is a clostridium shuttle plasmid. There are a series of plasmids from the pMTL80000 system. They differ in their Gram+ origins of replication: pMTL82151 (pBP1 from C. botulinum); pMTL83151 (pCB102 from C. butyricum); pMTL84151 (pCD6 from C. difficile); and pMTL85141 (pIM13 from Bacillus subtilis).
- Vector Name:
- pMTL84151
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6297 bp
- Type:
- Shuttle vector
- Replication origin:
- ori
- Source/Author:
- Heap JT, Pennington OJ, Cartman ST, Minton NP.
- Growth Temperature:
- 37℃
pMTL84151 vector Map
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add 20 μl of sterile water to dissolve the DNA.
3. Close the tube and incubate for 10 minutes at room temperature.
4. Briefly vortex the tube and then do a quick spin to concentrate the liquid at the bottom. Speed is less than 5000×g.
5. Store the plasmid at -20 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
References
- Heap JT, Pennington OJ, Cartman ST, Minton NP. A modular system for Clostridium shuttle plasmids. J Microbiol Methods. 2009;78(1):79-85.
pMTL84151 vector Sequence
LOCUS V004414 6297 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004414
VERSION V004414
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6297)
AUTHORS Heap JT, Pennington OJ, Cartman ST, Minton NP.
TITLE A modular system for Clostridium shuttle plasmids
JOURNAL J. Microbiol. Methods 78 (1), 79-85 (2009)
PUBMED 19445976
REFERENCE 2 (bases 1 to 6297)
AUTHORS Pennington OJ, Heap JT, Cartman ST, Minton NP.
TITLE Direct Submission
JOURNAL Submitted (02-MAR-2009) Centre for Biomolecular Sciences, The
University of Nottingham, University Park, Nottingham NG7 2RD,
United Kingdom
REFERENCE 3 (bases 1 to 6297)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6297)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J.
Microbiol. Methods"; date: "2009"; volume: "78"; issue: "1"; pages:
"79-85"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-MAR-2009) Centre for Biomolecular Sciences, The University of
Nottingham, University Park, Nottingham NG7 2RD, United Kingdom"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6297
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 9..62
/note="terminator derived from CD0164 gene of Clostridium
difficile"
/regulatory_class="terminator"
primer_bind 63..79
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(199..215)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS 569..1216
/codon_start=1
/product="putative plasmid replication protein"
/label="putative plasmid replication protein"
/note="orfB"
/protein_id="ACR43904.1"
/translation="MNKKLEKPFVYKREYDLTGYDVEILQKYELEQAIYVYVGSSCAYN
MRARSSKWRYHIRTNNKSICCNIKNFIHNLELFYKMELKLSDNIINDKLYYSNIAEFEE
FETLEKAREVESTIISQYQFLDSINHMLKQKIILLSNKDSVLNITKNGNTNYLKVKNKY
IEKHKNKPIMRYHINCQFNTDGSVKSITQEFEPILELNKKNTLSRPSRVFLK"
CDS 1491..3128
/codon_start=1
/gene="repA"
/product="plasmid replication protein"
/label="repA"
/protein_id="ACR43903.1"
/translation="MEQLDSKYKLKKFLMAVFRDGIGQGNNLIDNEYVRVFQNNKSNSK
QLELGEEFKEYSKTTFFKNIDDIVEFTFAKNIYYENTFFNLCTTDGKAGTNENLINRYA
LGFDFDKKELGQGFNYKDIINLFTKIGLHYHILVDSGNGFHVYVLINKTNNIKLVSEVT
NTLINKLGADKQANLSTQVLRVPYTYNIKNTTKQVKIIHQDKNIYRYDIEKLAKKYCKD
VKTVGNTNTKYILDSKLPNCIVDILKNGSKDGHKNLDLQKIVVTLRLRNKSLSQVISVA
REWNYISQNSLSNSELEYQVKYMYEKLKTVNFGCTGCEFNSDCWNKIESDFIYSDEDTL
FNMPHKHSKDLKYKNRKGVKIMTGNQLFIYNVLLNNKDRELNIDDIMELITYKRKKKVK
NIVMSEKTLRETLKELQHNDYITKTKGVTKLGIKDTYNVKEVRCNIDKQYTISYFVTMA
VIWGIISTEELRLYTHMRYKQDLLVKDDKIKGNILRINQEELAKDLGVTQQRISNMIES
LLDTKILDVWETKINDRGFMYYTYRLNK"
gene 1491..3128
/gene="repA"
/label="repA"
CDS 3941..4561
/gene="catP"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from Clostridium
perfringens. Accession#: P26826"
rep_origin 4741..5329
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
oriT 5650..5759
/label="oriT"
/note="incP origin of transfer"
CDS 5792..6160
/label="traJ"
/note="oriT-recognizing protein"