pHelper vector (V005569)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V005569	pHelper	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

The pHelper is a Adeno-associated viruses (AAV) helper plasmid. Specific gene products can be provided by helper plasmids (e.g., pHelper) and specific packaging cell lines (e.g., HEK293 cells) during AAV production.

Vector Name:: pHelper

Antibiotic Resistance:: Ampicillin

Length:: 11635 bp

Type:: Cloning vector

Replication origin:: ori

Source/Author:: Grafsky AJ III.

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

pHelper vector Map

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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

Park S, Shin S, Lee H, Jang JH, Lee GM. Enhancing the production of adeno-associated virus (AAV)2 and AAV9 with high full capsid ratio in HEK293 cells through design-of-experiment optimization of triple plasmid ratio. Biotechnol J. 2024;19(3):e2300667. doi:10.1002/biot.202300667

pHelper vector Sequence

LOCUS       V005569                11635 bp    DNA     circular SYN 18-DEC-2018
DEFINITION  Exported.
ACCESSION   V005569
VERSION     V005569
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 11635)
  AUTHORS   Grafsky AJ III.
  TITLE     pHelper for use in the AAV Helper-Free System
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 11635)
  AUTHORS   Grafsky AJ III.
  TITLE     Direct Submission
  JOURNAL   Submitted (16-APR-2001) Technical Services, Stratagene, 11011 N.
            Torrey Pines Rd., La Jolla, CA 92037, USA
REFERENCE   3  (bases 1 to 11635)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 11635)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName:
            "Unpublished"
            SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
            (16-APR-2001) Technical Services, Stratagene, 11011 N. Torrey Pines
            Rd., La Jolla, CA 92037, USA"
            SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..11635
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(261..1847)
                     /gene="DBP"
                     /label="DNA-binding protein"
                     /note="DNA-binding protein from Human adenovirus C serotype
                     2. Accession#: P03264"
     CDS             1876..4290
                     /gene="L4"
                     /label="Shutoff protein"
                     /note="Shutoff protein from Human adenovirus C serotype 2.
                     Accession#: P24932"
     CDS             5647..6030
                     /note="Early 4 ORF1 protein from Human adenovirus C
                     serotype 2. Accession#: P03242"
                     /label="Early 4 ORF1 protein"
     CDS             6475..6822
                     /note="Early 4 ORF3 protein from Human adenovirus C
                     serotype 2. Accession#: P03241"
                     /label="Early 4 ORF3 protein"
     CDS             7103..7984
                     /note="Early 4 ORF6 protein from Human adenovirus C
                     serotype 2. Accession#: P03239"
                     /label="Early 4 ORF6 protein"
     misc_feature    complement(8537..9280)
                     /label="similar to Adenovirus-5 VA"
                     /note="similar to Adenovirus-5 VA"
     rep_origin      complement(9423..10011)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(10185..11042)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(11043..11147)
                     /label="AmpR promoter"
     rep_origin      complement(11173..11628)
                     /direction=LEFT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"