Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V008474 | pCL | 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
- Vector Name:
- pCL
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3595 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Uhde K, Kerschbaumer RJ, Koenig R, Hirschl S, Lemaire O, Boonham N, Roake W, Himmler G.
pCL 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.
pCL vector Sequence
LOCUS 40924_10926 3595 bp DNA circular SYN 17-DEC-2018
DEFINITION Cloning vector pCL, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 3595)
AUTHORS Uhde K, Kerschbaumer RJ, Koenig R, Hirschl S, Lemaire O, Boonham N,
Roake W, Himmler G.
TITLE Improved detection of Beet necrotic yellow vein virus in a DAS ELISA
by means of antibody single chain fragments (scFv) which were
selected to protease-stable epitopes from phage display libraries
JOURNAL Arch. Virol. 145 (1), 179-185 (2000)
PUBMED 10664416
REFERENCE 2 (bases 1 to 3595)
AUTHORS Kerschbaumer RJ, Hirschl S, Himmler G.
TITLE Direct Submission
JOURNAL Submitted (02-AUG-1999) Strain Improvement, Institute of Applied
Microbiology, Muthgasse 18, Vienna, AT A-1190, Austria
REFERENCE 3 (bases 1 to 3595)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 3595)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Arch.
Virol."; date: "2000"; volume: "145"; issue: "1"; pages: "179-185"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(02-AUG-1999) Strain Improvement, Institute of Applied Microbiology,
Muthgasse 18, Vienna, AT A-1190, Austria"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..3595
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 96..200
/label=AmpR promoter
CDS 201..1058
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 1232..1820
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2108..2129
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2144..2174
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2182..2198
/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)."
primer_bind 2206..2222
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
sig_peptide 2274..2339
/label=pelB signal sequence
/note="leader peptide for secretion"
CDS 2373..2687
/codon_start=1
/label=hIg-kappa-CL
/note="Human immunoglobulin kappa light chain constant
region"
/translation="TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNA
LQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRG
E"
CDS 2691..2708
/codon_start=1
/label=6xHis
/note="6xHis affinity tag"
/translation="HHHHHH"
primer_bind complement(2727..2743)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 2954..3409
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"