Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V004330 | pNG4 | 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:
- pNG4
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9791 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Pisciotta A, Alduina R, Manteca A.
pNG4 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.
pNG4 vector Sequence
LOCUS V004330 9791 bp DNA circular SYN 18-DEC-2018
DEFINITION Exported.
ACCESSION V004330
VERSION V004330
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9791)
AUTHORS Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Pisciotta
A, Alduina R, Manteca A.
TITLE New PhiBT1 site-specific integrative vectors with neutral phenotype
in Streptomyces
JOURNAL Appl. Microbiol. Biotechnol. 100 (6), 2797-2808 (2016)
PUBMED 26758297
REFERENCE 2 (bases 1 to 9791)
AUTHORS Gonzalez-Quinonez N, Lopez-Garcia MT, Yague P, Rioseras B, Manteca
A.
TITLE Direct Submission
JOURNAL Submitted (15-APR-2015) Biologia Funcional, Universidad de Oviedo,
c/Julian Claveria s/n Facultad de Medicina, Oviedo 33006, Spain
REFERENCE 3 (bases 1 to 9791)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 9791)
AUTHORS .
TITLE Direct Submission
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
SGRef: number: 1; type: "Journal Article"; journalName: "Appl.
Microbiol. Biotechnol."; date: "2016"; volume: "100"; issue: "6";
pages: "2797-2808"
SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-APR-2015) Biologia Funcional, Universidad de Oviedo, c/Julian
Claveria s/n Facultad de Medicina, Oviedo 33006, Spain"
SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9791
/mol_type="other DNA"
/organism="synthetic DNA construct"
CDS complement(120..977)
/label="AmpR"
/note="beta-lactamase"
promoter complement(978..1041)
/label="AmpR promoter"
rep_origin 1310..1898
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2186..2207
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2222..2252
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 2260..2276
/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 2284..2300
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
terminator 2463..2511
/label="fd terminator"
/note="central terminator from bacteriophage fd (Otsuka and
Kunisawa, 1982)"
regulatory complement(2822..2825)
/regulatory_class="ribosome_binding_site"
oriT 5097..5206
/label="oriT"
/note="incP origin of transfer"
CDS 5239..5607
/label="traJ"
/note="oriT-recognizing protein"
CDS complement(6323..7318)
/gene="hyg"
/label="Hygromycin-B 7''-O-kinase"
/note="Hygromycin-B 7''-O-kinase from Streptomyces
hygroscopicus. Accession#: P09979"
CDS complement(8003..9313)
/codon_start=1
/gene="SCO4849"
/product="SCO4849 protein"
/label="SCO4849"
/protein_id="AML61769.1"
/translation="MVVVFVLVALLMVGVLVTANWYVWRRLFRDTTRAPGPVRRIGAAV
IAGGWLLAVGALVAERAGAPFWLQRVLAWPGFLWLALSIYLLLAVLAGEVVRPLLRRFL
ERRAAARRTAGAEPSVTPAADTSRIPAGTAPPKTPEAPETPEAPGTEALAAQGSPLAVP
SRRLFVSRVVAGAAAAAAVGTVGYGTYGVLSGPKVKRVTVPLAKLPRAAHGFRIAVVSD
IHLGPVLGRGFAQQVVDTINSTQPDLIAVVGDLVDGSVKDLGPAAAPLARLTARHGAYF
VTGNHEYFSGAEQWVAEVRRLGLLPLENARTELPHFDLAGVNDVAGEDEGQGPDYDRAL
GDRDRSRACVLLAHQPVQIHDAVDHGVDLQLSGHTHGGQLWPGNLIAGAANPTLAGLER
YGDTQLYVSRGAGAWGPPTRVGAPSDITVIELASRQA"
gene complement(8003..9313)
/gene="SCO4849"
/label="SCO4849"
regulatory complement(9368..9790)
/regulatory_class="promoter"