Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V010259 | pKLAC1 | 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
- Vector Name:
- pKLAC1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9091 bp
- Type:
- Yeast Plasmids
- Replication origin:
- ori
- Source/Author:
- Colussi PA, Taron CH.
- Copy Number:
- High copy number
- Promoter:
- LAC4 (3')
- Growth Strain(s):
- DH10B
- Growth Temperature:
- 37℃
pKLAC1 vector 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.
pKLAC1 vector Sequence
LOCUS 40924_26849 9091 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector pKLAC1, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9091)
AUTHORS Colussi PA, Taron CH.
TITLE Kluyveromyces lactis LAC4 promoter variants that lack function in
bacteria but retain full function in K. lactis
JOURNAL Appl. Environ. Microbiol. 71 (11), 7092-7098 (2005)
PUBMED 16269745
REFERENCE 2 (bases 1 to 9091)
AUTHORS Colussi PA, Taron CH.
TITLE Direct Submission
JOURNAL Submitted (18-MAR-2005) Research Department, New England Biolabs, 32
Tozer Road, Beverly, MA 01915, USA
REFERENCE 3 (bases 1 to 9091)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9091)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2005"; volume: "71"; issue: "11";
pages: "7092-7098"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(18-MAR-2005) Research Department, New England Biolabs, 32 Tozer
Road, Beverly, MA 01915, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9091
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 14..334
/note="Kluyveromyces lactis alpha mating factor pre-pro
secretion leader"
misc_feature 290..338
/label=multiple cloning site
/note="multiple cloning site"
terminator 355..973
/label=LAC4 terminator
/note="transcription terminator for Kluyveromyces lactis
LAC4"
promoter 974..1676
/label=ADH1 promoter
/note="promoter for alcohol dehydrogenase 1"
CDS 1697..3340
/label=amdS
/note="acetamidase from Aspergillus nidulans"
promoter 3979..4635
/label=LAC4 promoter (5')
/note="5' portion of the Kluyveromyces lactis LAC4
promoter"
promoter complement(4652..4670)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(4684..4700)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4708..4724)
/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)."
promoter complement(4732..4762)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4777..4798)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5086..5674)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5848..6705)
/label=AmpR
/note="beta-lactamase"
promoter complement(6706..6810)
/label=AmpR promoter
rep_origin 6837..7292
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 7433..7449
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 7463..8991
/label=LAC4 promoter (3')
/note="3' portion of a modified Kluyveromyces lactis LAC4
promoter"
protein_bind 8997..9013
/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)."