Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V009471 | pAP3neo | 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:
- pAP3neo
- Antibiotic Resistance:
- Ampicillin
- Length:
- 5350 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Kobori M, Ikeda Y, Nara H, Kato M, Kumegawa M, Nojima H, Kawashima H.
pAP3neo vector Map
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.
pAP3neo vector Sequence
LOCUS Amp;_beta-lactam 5350 bp DNA circular SYN 18-DEC-2018
DEFINITION Cloning vector pAP3neo DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS Amp; beta-lactamase; neomycin phosphotransferase; neo
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5350)
AUTHORS Kobori M, Ikeda Y, Nara H, Kato M, Kumegawa M, Nojima H, Kawashima
H.
TITLE Large scale isolation of osteoclast-specific genes by an improved
method involving the preparation of a subtracted cDNA library
JOURNAL Genes Cells 3 (7), 459-475 (1998)
PUBMED 9753427
REFERENCE 2 (bases 1 to 5350)
AUTHORS Nojima H.
TITLE Direct Submission
JOURNAL Submitted (30-MAY-1997) Hiroshi Nojima, Research Institute for
Microbial Diseases, Osaka University, Department of Molecular
Genetics; Yamadaoka 3-1, Suita, Osaka 565-0871, Japan
(E-mail:snj-0212@biken.osaka-u.ac.jp, Tel:81-6-6875-3980,
Fax:81-6-6875-5192)
REFERENCE 3 (bases 1 to 5350)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 5350)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Genes
Cells"; date: "1998"; volume: "3"; issue: "7"; pages: "459-475"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(30-MAY-1997) Hiroshi Nojima, Research Institute for Microbial
Diseases, Osaka University, Department of Molecular Genetics";
volume: " Yamadaoka 3-1, Suita, Osaka 565-0871, Japan
(E-mail:snj-0212@biken.osaka-u.ac.jp, Tel:81-6-6875-3980, Fax";
pages: "81-6-6875-5192"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5350
/mol_type="other DNA"
/organism="synthetic DNA construct"
polyA_signal 43..177
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 47..268
/label=SV40 late polyadenylation signal
/note="SV40 late polyadenylation signal"
/regulatory_class="polyA_signal_sequence"
misc_feature 428..568
/label=bom
/note="basis of mobility region from pBR322"
rep_origin complement(754..1342)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(1516..2373)
/label=AmpR
/note="beta-lactamase"
promoter complement(2374..2478)
/label=AmpR promoter
rep_origin complement(2504..2959)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 3101..3117
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 3169..3466
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3596..4387
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
polyA_signal complement(4442..4576)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter 4654..4983
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
intron 5022..5118
/label=SV40 intron
/note="modified SV40 intron with splice donor and acceptor
sites"
promoter 5188..5206
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(5303..5321)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"