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pNFkB-Luc vector (V004337)

Price Information

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V004337 pNFkB-Luc In stock, instant shipping

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Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

NFkB reporter vector is utilized for detecting NFkB (Nuclear factor of kappa light polypeptide gene) transactivation in a cell-based assay.

Vector Name:
pNFkB-Luc
Antibiotic Resistance:
Ampicillin
Length:
5695 bp
Type:
Reporter vector
Replication origin:
ori
Source/Author:
Zheng C-F.
Selection Marker:
Fluc
Promoter:
minP

pNFkB-Luc vector Map

Copy Sequence View Map
pNFkB-Luc5695 bp60012001800240030003600420048005400AmpR promoterAmpRoribomSV40 poly(A) signalSV40 poly(A) signalNFkB response elementsminPluciferasesmall t intronSV40 T-ANTIGENSV40 poly(A) signal

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.

pNFkB-Luc vector Sequence

LOCUS       40924_33133        5695 bp DNA     circular SYN 16-JAN-2024
DEFINITION  Reporter vector pNFkB-Luc, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5695)
  AUTHORS   Zheng C-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (11-MAR-1998) Technical Services, Stratagene, 11011 N. 
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   2  (bases 1 to 5695)
  AUTHORS   Zheng C-F.
  TITLE     Direct Submission
  JOURNAL   Submitted (20-JAN-1999) Technical Services, Stratagene, 11011 N. 
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 5695)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (11-MAR-1998) Technical Services, Stratagene, 11011 N. Torrey Pines 
            Rd., La Jolla, CA 92037, USA"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (20-JAN-1999) Technical Services, Stratagene, 11011 N. Torrey Pines 
            Rd., La Jolla, CA 92037, USA"
FEATURES             Location/Qualifiers
     source          1..5695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        32..136
                     /label=AmpR promoter
     CDS             137..994
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      1168..1755
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_feature    complement(1934..2074)
                     /label=bom
                     /note="basis of mobility region from pBR322"
     polyA_signal    2313..2445
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    2539..2671
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    2680..2746
                     /label=NFkB response elements
     promoter        2757..2788
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     CDS             2815..4464
                     /label=luciferase
                     /note="firefly luciferase"
     intron          4598..4663
                     /label=small t intron
                     /note="SV40 (simian virus 40) small t antigen intron"
     CDS             4854..5210
                     /codon_start=1
                     /label=SV40 T-ANTIGEN
                     /translation="MLCLVIELLLALLFTTTKEKAALLYKKIMEKYSVTFISRHNSYNH
                     NILFFLTPHRHRVSAINNYAQKLCTFSFLICKGVNKEYLMYSALTRDHNQPYHICRGFT
                     CFKKPPTPPPEPET"
     polyA_signal    5233..5367
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"