Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012834 | pMTL-SC7315 | In stock, instant shipping |
Buy one, get one free! |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
Plasmid vector pMTL-SC7315, used for allele exchange in C. difficile 630. Note that pMTL-SC7215 and pMTL-SC7315 are identical except that they have the pBP1 and pCB102 Gram-positive replicons, respectively (between AscI and FseI restriction sites).
- Vector Name:
- pMTL-SC7315
- Antibiotic Resistance:
- Chloramphenicol
- Length:
- 6000 bp
- Type:
- C. difficile
- Replication origin:
- ori
- Source/Author:
- Cartman ST, Kelly ML, Heeg D, Heap JT , Minton NP
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
pMTL-SC7315 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.
pMTL-SC7315 vector Sequence
LOCUS V012834 6000 bp DNA circular SYN 04-JAN-2024
DEFINITION Exported.
ACCESSION V012834
VERSION V012834
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 6000)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6000
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 63..79
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(254..278)
/label="lac operator"
/note="The lac repressor binds to the lac operator to
inhibit transcription in E. coli. This inhibition can be
relieved by adding lactose or
isopropyl-beta-D-thiogalactopyranoside (IPTG)."
RBS 297..305
/label="Shine-Dalgarno sequence"
/note="full consensus sequence for ribosome-binding sites
upstream of start codons in E. coli; complementary to a
region in the 3' end of the 16S rRNA (Chen et al., 1994)"
CDS 318..1598
/label="codA"
/note="E. coli cytosine deaminase"
primer_bind complement(1723..1739)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
CDS 2655..2972
/codon_start=1
/gene="repH"
/product="putative plasmid replication protein"
/label="repH"
/protein_id="ACR43897.1"
/translation="MKRRAYMKTCKNCKEFIKDTEICKIHSLMIHDKTVATYCSKYNES
RCLHKGKVQCINCSKMNRYGWCAIKMRCFTEEEQKKERTCIKYYARSFKKAHVKKSKKK
K"
gene 2655..2972
/gene="repH"
/label="repH"
CDS 3644..4264
/gene="catP"
/label="Chloramphenicol acetyltransferase"
/note="Chloramphenicol acetyltransferase from Clostridium
perfringens. Accession#: P26826"
rep_origin 4444..5032
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
oriT 5353..5462
/label="oriT"
/note="incP origin of transfer"
CDS 5495..5863
/label="traJ"
/note="oriT-recognizing protein"