HA Nprl3 pRK5 vector (V017351)

Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V017351	HA Nprl3 pRK5	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:: HA Nprl3 pRK5

Antibiotic Resistance:: Ampicillin

Length:: 6475 bp

Type:: Protein expression

Replication origin:: ori

Host:: Mammalian cells

Promoter:: SV40

5' Primer:: CMV-F

Growth Temperature:: 37℃

HA Nprl3 pRK5 vector Vector Map

View Map in NovoBuilder

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.

HA Nprl3 pRK5 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V017351                 6475 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V017351
VERSION     V017351
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 6475)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6475
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        143..173
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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     205..221
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     enhancer        250..629
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1064..1082
                     /label="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     CDS             1211..1237
                     /label="HA"
                     /note="HA (human influenza hemagglutinin) epitope tag"
     CDS             1268..2974
                     /gene="NPRL3"
                     /label="GATOR1 complex protein NPRL3"
                     /note="GATOR1 complex protein NPRL3 from Homo sapiens.
                     Accession#: Q12980"
     polyA_signal    3024..3158
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        3227..3584
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3604..3620)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      3833..4288
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4570..4674
                     /label="AmpR promoter"
     CDS             4675..5532
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      5706..6294
                     /direction=RIGHT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"