Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V006653 | pCMV-Neo-Bam APC | 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
- Vector Name:
- pCMV-Neo-Bam APC
- Antibiotic Resistance:
- Ampicillin
- Length:
- 15521 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- Low Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- Bglob-intron-F
- 3' Primer:
- Bglob-pA-R (5'-ttttggcagagggaaaaaga-3')
pCMV-Neo-Bam APC 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.
pCMV-Neo-Bam APC vector Sequence
LOCUS V006653 15521 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V006653
VERSION V006653
KEYWORDS pCMV-Neo-Bam APC
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 15521)
AUTHORS Morin PJ, Sparks AB, Korinek V, Barker N, Clevers H, Vogelstein B,
Kinzler KW
TITLE Activation of beta-catenin-Tcf signaling in colon cancer by
mutations in beta-catenin or APC.
JOURNAL Science. 1997 Mar 21. 275(5307):1787-90.
PUBMED 9065402
REFERENCE 2 (bases 1 to 15521)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 15521)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Science.
1997 Mar 21. 275(5307):1787-90."
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..15521
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 381..584
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 581..605
/label="LNCX"
/note="Human CMV promoter, forward primer"
intron 696..1268
/label="beta-globin intron"
/note="intron from rabbit beta-globin gene"
CDS 1374..9902
/gene="APC"
/label="Adenomatous polyposis coli protein"
/note="Adenomatous polyposis coli protein from Homo
sapiens. Accession#: P25054"
primer_bind complement(10443..10462)
/label="Bglob-pA-R"
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 10508..10563
/label="beta-globin poly(A) signal"
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(10562..10581)
/label="rbglobpA-R"
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
primer_bind complement(10921..10937)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(10921..10937)
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(10934..10956)
/label="M13/pUC Reverse"
/note="In lacZ gene"
protein_bind 10945..10961
/label="lac operator"
/bound_moiety="lac repressor encoded by lacI"
/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(10969..10999)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(11014..11035)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(11152..11169)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(11323..11911)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12085..12942)
/label="AmpR"
/note="beta-lactamase"
promoter complement(12943..13047)
/label="AmpR promoter"
primer_bind 13115..13133
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
primer_bind complement(13171..13193)
/label="pGEX 3'"
/note="pGEX vectors, reverse primer"
primer_bind 13293..13312
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
primer_bind 13521..13537
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
promoter 13948..14093
/label="HSV TK promoter"
/note="herpes simplex virus thymidine kinase promoter"
CDS 14150..14941
/label="NeoR/KanR"
/note="aminoglycoside phosphotransferase"
primer_bind complement(15138..15157)
/label="TK-pA-R"
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 15182..15229
/label="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"