Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012204 | pLenti-puro-ARID1A | 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:
- pLenti-puro-ARID1A
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13943 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Puromycin
- Copy Number:
- High Copy
- Promoter:
- RSV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-fwd
- 3' Primer:
- BGH-rev
pLenti-puro-ARID1A 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.
pLenti-puro-ARID1A vector Sequence
LOCUS V012204 13943 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V012204
VERSION V012204
KEYWORDS pLenti-puro-ARID1A
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13943)
AUTHORS Guan B, Wang TL, Shih IeM
TITLE ARID1A, a factor that promotes formation of SWI/SNF-mediated
chromatin remodeling, is a tumor suppressor in gynecologic cancers.
JOURNAL Cancer Res. 2011 Nov 1;71(21):6718-27. Epub 2011 Sep 7.
PUBMED 21900401
REFERENCE 2 (bases 1 to 13943)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13943)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cancer
Res."; date: "2011-11-1"; volume: "71(21)"; pages: "6718-27. Epub
2011 Sep 7"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13943
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 186..208
/label="M13/pUC Reverse"
/note="In lacZ gene"
primer_bind 205..221
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 205..221
/label="M13 Reverse"
/note="In lacZ gene. Also called M13-rev"
promoter 242..260
/label="T3 promoter"
/note="promoter for bacteriophage T3 RNA polymerase"
promoter 288..514
/label="RSV promoter"
/note="Rous sarcoma virus enhancer/promoter"
LTR 515..695
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 742..867
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1360..1593
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1778..1822
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1971..2012
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
enhancer 2101..2403
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 2404..2607
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
protein_bind 2609..2627
/label="tet operator"
/note="bacterial operator O2 for the tetR and tetA genes"
protein_bind 2630..2648
/gene="tetO"
/label="tet operator"
/bound_moiety="tetracycline repressor TetR"
/note="bacterial operator O2 for the tetR and tetA genes"
CDS 2732..9586
/gene="ARID1A"
/label="AT-rich interactive domain-containing protein 1A"
/note="AT-rich interactive domain-containing protein 1A
from Homo sapiens. Accession#: O14497"
CDS 9614..9655
/label="V5 tag"
/note="epitope tag from simian virus 5"
CDS 9665..9682
/label="6xHis"
/note="6xHis affinity tag"
primer_bind complement(9705..9722)
/label="BGH-rev"
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
primer_bind complement(9734..9754)
/label="pBABE 3'"
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 9739..10068
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 9919..10054
/label="SV40 ori"
/note="SV40 origin of replication"
CDS 10078..10674
/label="PuroR"
/note="puromycin N-acetyltransferase"
LTR 10749..10982
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 11054..11188
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin 11215..11350
/label="SV40 ori"
/note="SV40 origin of replication"
promoter complement(11371..11389)
/label="T7 promoter"
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(11399..11416)
/label="M13 Forward"
/note="In lacZ gene. Also called M13-F20 or M13 (-21)
Forward"
primer_bind complement(11399..11415)
/label="M13 fwd"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(11408..11430)
/label="M13/pUC Forward"
/note="In lacZ gene"
rep_origin 11557..12012
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 12038..12142
/label="AmpR promoter"
CDS 12143..13000
/label="AmpR"
/note="beta-lactamase"
rep_origin 13174..13762
/direction=RIGHT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 13916..13933
/label="L4440"
/note="L4440 vector, forward primer"