pAAV2/7 vector (Cat. No.: V012817)
Note: AAV packaging plasmid, expressing Rep2 and Cap7
- Name:
- pAAV2/7
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7333 bp
- Type:
- Viral Expression & Packaging Vectors
- Replication origin:
- ori
- Promoter:
- Truncated P5
- Growth Strain(s):
- DH5alpha
- Growth Temperature:
- 37℃
Resources
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.
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.
pAAV2/7 vector (Cat. No.: V012817) Sequence
LOCUS pAAV2-7 7333 bp DNA circular SYN 09-MAY-2026
DEFINITION Exported.
ACCESSION V012817
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7333)
TITLE Direct Submission
REFERENCE 2 (bases 1 to 7333)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7333
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 12..28
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 38..56
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind 86..102
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
promoter 121..156
/label=Truncated P5
/label=Truncated\P5
/note="/vntifkey=29"
CDS 157..2019
/gene="Rep78"
/label=Protein Rep78
/note="Protein Rep78 from Adeno-associated virus 2 (isolate
Srivastava/1982). Accession#: Q89268"
CDS 2039..4252
/codon_start=1
/label=Capsid protein AAV7
/translation="MAADGYLPDWLEDNLSEGIREWWDLKPGAPKPKANQQKQDNGRGL
VLPGYKYLGPFNGLDKGEPVNAADAAALEHDKAYDQQLKAGDNPYLRYNHADAEFQERL
QEDTSFGGNLGRAVFQAKKRVLEPLGLVEEGAKTAPAKKRPVEPSPQRSPDSSTGIGKK
GQQPARKRLNFGQTGDSESVPDPQPLGEPPAAPSSVGSGTVAAGGGAPMADNNEGADGV
GNASGNWHCDSTWLGDRVITTSTRTWALPTYNNHLYKQISSETAGSTNDNTYFGYSTPW
GYFDFNRFHCHFSPRDWQRLINNNWGFRPKKLRFKLFNIQVKEVTTNDGVTTIANNLTS
TIQVFSDSEYQLPYVLGSAHQGCLPPFPADVFMIPQYGYLTLNNGSQSVGRSSFYCLEY
FPSQMLRTGNNFEFSYSFEDVPFHSSYAHSQSLDRLMNPLIDQYLYYLARTQSNPGGTA
GNRELQFYQGGPSTMAEQAKNWLPGPCFRQQRVSKTLDQNNNSNFAWTGATKYHLNGRN
SLVNPGVAMATHKDDEDRFFPSSGVLIFGKTGATNKTTLENVLMTNEEEIRPTNPVATE
EYGIVSSNLQAANTAAQTQVVNNQGALPGMVWQNRDVYLQGPIWAKIPHTDGNFHPSPL
MGGFGLKHPPPQILIKNTPVPANPPEVFTPAKFASFITQYSTGQVSVEIEWELQKENSK
RWNPEIQYTSNFEKQTGVDFAVDSQGVYSEPRPIGTRYLTRNL"
CDS 4094..4105
/label=WELQut site
/note="WELQut protease recognition and cleavage site"
misc_feature 4342..4473
/label=P5 promoter
/label=P5\\promoter
/note="/vntifkey=21"
promoter complement(4557..4575)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(4596..4612)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4620..4636)
/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(4644..4674)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(4689..4710)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(4998..5586)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(join(5757..6548,6549..6617))
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
CDS complement(5760..6617)
/label=AmpR
/note="beta-lactamase"
promoter complement(6618..6722)
/label=AmpR promoter
rep_origin complement(6748..7203)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"