Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004184 | pP43NMK | 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:
- pP43NMK
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7680 bp
- Type:
- Shuttle expression-secretion vector
- Replication origin:
- ori
- Source/Author:
- Zhang XZ, Cui ZL, Hong Q, Li SP.
pP43NMK 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.
pP43NMK vector Sequence
LOCUS 40924_34022 7680 bp DNA circular SYN 18-DEC-2018
DEFINITION Shuttle expression-secretion vector pP43NMK, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7680)
AUTHORS Zhang XZ, Cui ZL, Hong Q, Li SP.
TITLE High-level expression and secretion of methyl parathion hydrolase in
Bacillus subtilis WB800
JOURNAL Appl. Environ. Microbiol. 71 (7), 4101-4103 (2005)
PUBMED 16000826
REFERENCE 2 (bases 1 to 7680)
AUTHORS Zhang X-Z., Cui Z-L., Li S-P.
TITLE Direct Submission
JOURNAL Submitted (25-OCT-2005) Department of Microbiology, College of Life
Sciences, Nanjing Agricultural University, MOA Key Lab of
Microbiological Engineering of Agricultural Environment, #6 Tongwei
Road, Weigang, Nanjing, Jiangsu 210095, P. R. China
REFERENCE 3 (bases 1 to 7680)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7680)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Environ. Microbiol."; date: "2005"; volume: "71"; issue: "7"; pages:
"4101-4103"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(25-OCT-2005) Department of Microbiology, College of Life Sciences,
Nanjing Agricultural University, MOA Key Lab of Microbiological
Engineering of Agricultural Environment, #6 Tongwei Road, Weigang,
Nanjing, Jiangsu 210095, P. R. China"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7680
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 379..395
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
CDS 814..1815
/label=repB
/note="RepB replication protein"
CDS 1987..2748
/codon_start=1
/gene="km"
/product="Km"
/label=km
/protein_id="ABD24446.1"
/translation="MNGPIIMTREERMKIVHEIKERILDKYGDDVKAIGVYGSLGRQTD
GPYSDIEMMCVMSTEEAEFSHEWTTGEWKVEVNFDSEEILLDYASQVESDWPLTHGQFF
SILPIYDSGGYLEKVYQTAKSVEAQTFHDAICALIVEELFEYAGKWRNIRVQGPTTFLP
SLTVQVAMAGAMLIGLHHRICYTTSASVLTEAVKQSDLPSGYDHLCQFVMSGQLSDSEK
LLESLENFWNGIQEWTERHGYIVDVSKRIPF"
gene 1987..2748
/gene="km"
/label=km
CDS 2965..3216
/codon_start=1
/gene="ble"
/product="Ble"
/label=ble
/protein_id="ABD24447.1"
/translation="MRMLQSIPALPVGDIKKSIGFYWDKVGFTLVHHEDGFAVLMCNEV
RIHLWEASDEGWRLVVMIHRFVQVRSRLLLVLLVAALK"
gene 2965..3216
/gene="ble"
/label=ble
regulatory 4367..4384
/note="-35 and -10 signal of promoter controlled by sigma
B"
/regulatory_class="minus_35_signal"
regulatory 4386..4391
/note="-35 and -10 signal of promoter controlled by sigma
A"
/regulatory_class="minus_35_signal"
regulatory 4400..4409
/note="-35 and -10 signal of promoter controlled by sigma
B"
/regulatory_class="minus_10_signal"
regulatory 4409..4414
/note="-35 and -10 signal of promoter controlled by sigma
A"
/regulatory_class="minus_10_signal"
mRNA 4419..5434
/gene="mpd"
/product="MPH"
/label=mpd mRNA
gene 4419..5434
/gene="mpd"
/label=mpd
regulatory 4441..4450
/gene="mpd"
/regulatory_class="ribosome_binding_site"
CDS 4460..5434
/codon_start=1
/gene="mpd"
/product="MPH"
/label=mpd
/note="methyl parathion hydrolase; includes NprB signal
peptide"
/protein_id="ABD24448.1"
/translation="MRNLTKTSLLLAGLCTAAQMVFVTHASAAAPQVRTSAPGYYRMLL
GDFEITALSDGTVALPVDKRLNQPAPKTQSALAKSFQKAPLETSVTGYLVNTGSKLVLV
DTGAAGLFGPTLGRLAANLKAAGYQPEQVDEIYITHMHPDHVGGLMVGEQLAFPNAVVR
ADQKEADFWLSQTNLDKAPDDESKGFFKGAMASLNPYVKAGKFKPFSGNTDLVPGIKAL
ASHGHTPGHTTYVVESQGQKLALLGDLILVAAVQFDDPSVTNQLDIDGKSAAVERKKAF
ADAAKGGYLIAASHLPFPGIGHIRAEGKGYRFVPVNYSVVNPK"
sig_peptide 4460..4546
/gene="mpd"
/note="from NprB"
primer_bind complement(5459..5475)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5483..5499)
/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(5507..5537)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5552..5573)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5861..6449)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6623..7480)
/label=AmpR
/note="beta-lactamase"
promoter complement(7481..7585)
/label=AmpR promoter