Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007264 | pc DNA FRT TO- APPSwed/Ind | 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:
- pc DNA FRT TO- APPSwed/Ind
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7225 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Promoter:
- CMV
- Cloning Method:
- Gibson Cloning
pc DNA FRT TO- APPSwed/Ind 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.
pc DNA FRT TO- APPSwed/Ind vector Sequence
LOCUS V007264 7225 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007264
VERSION V007264
KEYWORDS pc DNA FRT TO- APPSwed/Ind
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 7225)
TITLE Radenovic Lab plasmids
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7225)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7225)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7225
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 97..476
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 477..680
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 682..700
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 703..721
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
primer_bind 731..755
/label="LNCX"
/note="Human CMV promoter, forward primer"
CDS complement(1217..1228)
/codon_start=1
/gene="cspA"
/product="translation enhancing element for E. coli (Qing
et al., 2004)"
/label="TEE"
/translation="NHKV"
CDS 1708..1728
/label="7xHis"
/note="6xHis affinity tag"
CDS 2678..2797
/gene="APP"
/label="Amyloid-beta precursor protein"
/note="Amyloid-beta precursor protein from Felis catus.
Accession#: P86906"
polyA_signal 3045..3269
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
primer_bind complement(3402..3421)
/label="F1ori-R"
/note="F1 origin, reverse primer"
protein_bind 3553..3600
/label="FRT"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
CDS 3608..4627
/label="HygR"
/note="aminoglycoside phosphotransferase from E. coli"
polyA_signal 4760..4893
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(4930..4946)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(4954..4970)
/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(4978..5008)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(5023..5044)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5161..5178)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(5332..5920)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6094..6951)
/label="AmpR"
/note="beta-lactamase"
promoter complement(6952..7056)
/label="AmpR promoter"
primer_bind complement(7131..7150)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"