pNV18 vector (V004250)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V004250	pNV18	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

pNV18 is created by inserting a 1.8 kb DNA fragment containing the pAL5000 origin of replication into the unique NheI site of either pK18. Key features of pNV18 include: 1. Compact size. 2. Multiple cloning sites: The latest version includes sites for HindIII, SphI, PstI, SalI, HincII, XbaI, BamHI, KpnI, and EcoRI. 3. The lacZ gene (α fragment) facilitates blue-white selection in E. coli. 4. The aph gene from Tn5 provides kanamycin and neomycin resistance in E. coli and Nocardia species. 5. Broad host range: Effective in N. farcinica, N. asteroides, and various mycobacteria.

Vector Name:: pNV18

Antibiotic Resistance:: Kanamycin

Length:: 4411 bp

Type:: Shuttle vector

Replication origin:: ori

Source/Author:: Chiba K, Hoshino Y, Ishino K, Kogure T, Mikami Y, Uehara Y, Ishikawa J.

Growth Strain(s):: Top10

Growth Temperature:: 37℃

pNV18 vector Map

Copy Sequence View 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.

References

Kan J, Peng T, Huang T, Xiong G, Hu Z. NarL, a Novel Repressor for CYP108j1 Expression during PAHs Degradation in Rhodococcus sp. P14. Int J Mol Sci. 2020 Feb 1;21(3):983. doi: 10.3390/ijms21030983. PMID: 32024188; PMCID: PMC7037279.

pNV18 vector Sequence

LOCUS       40924_33652        4411 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Shuttle vector pNV18 DNA, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4411)
  AUTHORS   Chiba K, Hoshino Y, Ishino K, Kogure T, Mikami Y, Uehara Y, Ishikawa
            J.
  TITLE     Construction of a pair of practical Nocardia-Escherichia coli 
            shuttle vectors
  JOURNAL   Jpn. J. Infect. Dis. 60 (1), 45-47 (2007)
  PUBMED    17314425
REFERENCE   2  (bases 1 to 4411)
  AUTHORS   Ishikawa J, Chiba K.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-JUL-2006) Contact:Jun Ishikawa National Institute of 
            Infectious Diseases, Department of Bioactive Molecules; 1-23-1 
            Toyama, Shinjuku, Tokyo 162-8640, Japan URL 
            :http://nocardia.nih.go.jp/
REFERENCE   3  (bases 1 to 4411)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 4411)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Jpn. J. 
            Infect. Dis."; date: "2007"; volume: "60"; issue: "1"; pages: 
            "45-47"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-JUL-2006) Contact:Jun Ishikawa National Institute of Infectious 
            Diseases, Department of Bioactive Molecules; 1-23-1 Toyama, 
            Shinjuku, Tokyo 162-8640, Japan URL :http://nocardia.nih.go.jp/"
COMMENT     SGRef: number: 3; type: "Journal Article"
COMMENT     On Dec 18, 2007 this sequence version replaced AB267085.1.
FEATURES             Location/Qualifiers
     source          1..4411
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     152..168
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    complement(172..228)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(238..254)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(262..278)
                     /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(286..316)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(331..352)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(640..1228)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(1515..2306)
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    2662..4411
                     /label=derived from pAL5000
                     /note="derived from pAL5000"
     CDS             complement(2746..3093)
                     /codon_start=1
                     /gene="repB"
                     /product="replication protein RepB"
                     /label=repB
                     /protein_id="BAF02356.2"
                     /translation="MSDGYSDGYNRQPTVRKKRRVTAAEGARITGLSERHVVRLVAQER
                     SEWLAEQAARRERIRAYHDDEGHSWPQTAKHFGLHLDTVKRLGYRARKERAAEQEAAQK
                     AHNEADNPPLF"
     gene            complement(2746..3093)
                     /gene="repB"
                     /label=repB
     CDS             complement(3090..4016)
                     /codon_start=1
                     /gene="repA"
                     /product="replication protein RepA"
                     /label=repA
                     /protein_id="BAF02357.1"
                     /translation="MSHVADEFEQLWLPYWPLASDDLLEGIYRQSRASALGRRYIEANP
                     TALANLLVVDVDHPDAALRALSARGSHPLPNAIVGNRANGHAHAVWALNAPVPRTEYAR
                     RKPLAYMAACAEGLRRAVDGDRSYSGLMTKNPGHIAWETEWLHSDLYTLSHIEAELGAN
                     MPPPRWRQQTTYKAAPTPLGRNCALFDSVRLWAYRPALMRIYLPTRNVDGLGRAIYAEC
                     HARNAEFPCNDVCPGPLPDSEVRAIANSIWRWITTKSRIWADGIVVYEATLSARQSAIS
                     RKGAAARTAASTVARRAKSASAMEALL"
     gene            complement(3090..4016)
                     /gene="repA"
                     /label=repA