pAAV2/9n vector (V001257)

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Price Information

Cat No.	Plasmid Name	Availability	Add to cart
V001257	pAAV2/9n	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

pAAV2/9n is a AAV packaging plasmid expressing Rep/Cap genes.This is a vector developed by Dr. James M. Wilson.

Vector Name:: pAAV2/9n

Antibiotic Resistance:: Ampicillin

Length:: 7330 bp

Type:: AAV

Replication origin:: ori

Promoter:: Truncated P5

5' Primer:: KS primer

Growth Strain(s):: stbl3

Growth Temperature:: 37℃

pAAV2/9n 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

Hu Y, Liu L, Chen Y, et al. Cancer-cell-secreted miR-204-5p induces leptin signalling pathway in white adipose tissue to promote cancer-associated cachexia. Nat Commun. 2023;14(1):5179. Published 2023 Aug 24. doi:10.1038/s41467-023-40571-9

pAAV2/9n vector Sequence

LOCUS       Exported                7330 bp DNA     circular SYN 15-OCT-2024
DEFINITION  Exported.
ACCESSION   V001257
VERSION     .
KEYWORDS    pAAV2-9n
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7330)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 7330)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7330)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     137..153
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        163..181
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     211..227
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             282..2144
                     /gene="Rep78"
                     /label=Protein Rep78
                     /note="Protein Rep78 from Adeno-associated virus 2 (isolate
                     Srivastava/1982). Accession#: Q89268"
     CDS             2164..4374
                     /codon_start=1
                     /label=Cap9
                     /note="Capsid protein VP1 [Adeno-associated virus 9]"
                     /translation="MAADGYLPDWLEDNLSEGIREWWALKPGAPQPKANQQHQDNARGL
                     VLPGYKYLGPGNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLKYNHADAEFQERL
                     KEDTSFGGNLGRAVFQAKKRLLEPLGLVEEAAKTAPGKKRPVEQSPQEPDSSAGIGKSG
                     AQPAKKRLNFGQTGDTESVPDPQPIGEPPAAPSGVGSLTMASGGGAPVADNNEGADGVG
                     SSSGNWHCDSQWLGDRVITTSTRTWALPTYNNHLYKQISNSTSGGSSNDNAYFGYSTPW
                     GYFDFNRFHCHFSPRDWQRLINNNWGFRPKRLNFKLFNIQVKEVTDNNGVKTIANNLTS
                     TVQVFTDSDYQLPYVLGSAHEGCLPPFPADVFMIPQYGYLTLNDGSQAVGRSSFYCLEY
                     FPSQMLRTGNNFQFSYEFENVPFHSSYAHSQSLDRLMNPLIDQYLYYLSKTINGSGQNQ
                     QTLKFSVAGPSNMAVQGRNYIPGPSYRQQRVSTTVTQNNNSEFAWPGASSWALNGRNSL
                     MNPGPAMASHKEGEDRFFPLSGSLIFGKQGTGRDNVDADKVMITNEEEIKTTNPVATES
                     YGQVATNHQSAQAQAQTGWVQNQGILPGMVWQDRDVYLQGPIWAKIPHTDGNFHPSPLM
                     GGFGMKHPPPQILIKNTPVPADPPTAFNKDKLNSFITQYSTGQVSVEIEWELQKENSKR
                     WNPEIQYTSNYYKSNNVEFAVNTEGVYSEPRPIGTRYLTRNL"
     CDS             4216..4227
                     /label=WELQut site
                     /note="WELQut protease recognition and cleavage site"
     promoter        complement(4679..4697)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4718..4734)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4742..4758)
                     /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(4766..4796)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4811..4832)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(4949..4966)
                     /label=L4440
                     /note="L4440"
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5120..5708)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5882..6739)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6740..6844)
                     /label=AmpR promoter
     rep_origin      complement(6870..7325)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"

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