Price Information
| Cat No. | Plasmid Name | Availability | Buy one, get one free! (?) |
|---|---|---|---|
| V004050 | pPL451 | In stock, instant shipping |
Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.
Basic Vector Information
- Vector Name:
- pPL451
- Antibiotic Resistance:
- Ampicillin
- Length:
- 4249 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Love CA, Lilley PE, Dixon NE.
- Growth Strain(s):
- stbl3
- Growth Temperature:
- 37℃
pPL451 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.
pPL451 vector Sequence
LOCUS 40924_34879 4249 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pPL451 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4249)
AUTHORS Love CA, Lilley PE, Dixon NE.
TITLE Stable high-copy-number bacteriophage lambda promoter vectors for
overproduction of proteins in Escherichia coli
JOURNAL Gene 176 (1-2), 49-53 (1996)
PUBMED 8918231
REFERENCE 2 (bases 1 to 4249)
AUTHORS Yasuda S.
TITLE Direct Submission
JOURNAL Submitted (07-FEB-2006) Contact:Seiichi Yasuda National Institute of
Genetics, Microbial Genetics; Yata 1111, Mishima, Shizuoka 411-8540,
Japan
REFERENCE 3 (bases 1 to 4249)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 4249)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Gene 176
(1-2), 49-53 (1996)"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(07-FEB-2006) Contact:Seiichi Yasuda National Institute of Genetics,
Microbial Genetics; Yata 1111, Mishima, Shizuoka 411-8540, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..4249
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_feature 53..419
/label=partition(pSC101)
/note="partition(pSC101)"
promoter 474..578
/label=AmpR promoter
CDS 579..1436
/label=AmpR
/note="beta-lactamase"
rep_origin 1610..2198
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2486..2507
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2522..2552
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2560..2576
/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 2584..2600
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
CDS complement(2859..3569)
/label=lambda repressor
/note="phage lambda repressor"
regulatory 3617..3644
/label=lambda PR
/note="lambda PR"
/regulatory_class="promoter"
RBS 3658..3666
/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)"
regulatory 3831..3859
/label=lambda PL
/note="lambda PL"
/regulatory_class="promoter"
misc_feature 3980..4084
/label=polylinker
/note="polylinker"
RBS 3986..3994
/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)"
primer_bind complement(4092..4108)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"