Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V017403 | lenti SYN-SVI-DIO-dCas9-KRAB-MeCP2 | 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:
- lenti SYN-SVI-DIO-dCas9-KRAB-MeCP2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 13818 bp
- Type:
- Gene template
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- BleoR
- Promoter:
- hSyn
- 5' Primer:
- CMV-F
- Growth Temperature:
- 37℃
lenti SYN-SVI-DIO-dCas9-KRAB-MeCP2 vector 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.
lenti SYN-SVI-DIO-dCas9-KRAB-MeCP2 vector Sequence
LOCUS V017403 13818 bp DNA circular SYN 01-JAN-1980
DEFINITION Exported.
ACCESSION V017403
VERSION V017403
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 13818)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..13818
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..380
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 381..583
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 598..778
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 825..950
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1443..1676
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1861..1905
/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 2054..2095
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 2203..2320
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2365..2812
/label="hSyn promoter"
/note="human synapsin I promoter; confers neuron-specific
expression (Kugler et al., 2003)"
protein_bind complement(2825..2858)
/label="lox2272"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
compatible with each other, but incompatible with loxP or
loxN sites (Lee and Saito, 1988)."
protein_bind complement(2909..2942)
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS complement(4430..4450)
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS complement(4457..4522)
/label="3xFLAG"
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
protein_bind 4532..4565
/label="lox2272"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
compatible with each other, but incompatible with loxP or
loxN sites (Lee and Saito, 1988)."
protein_bind 4616..4649
/label="loxP"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
CDS 4914..7412
/label="Cas9(C)"
/note="C-terminal portion of Streptococcus pyogenes Cas9
(Zetsche et al., 2015)"
CDS 7521..7715
/label="KRAB"
/note="Kruppel-associated box (KRAB) transcriptional
repression domain from the human zinc finger protein ZNF10
(Margolin et al., 1994)"
CDS 7749..7769
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 7779..8636
/label="MeCP2"
/note="transcriptional repression domain from rat
methyl-CpG-binding protein 2 (Nan et al., 1997)"
misc_feature 8673..9261
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 9333..9566
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 9598..9822
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
promoter 10310..10639
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 10687..10734
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 10753..11124
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 11257..11390
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
promoter complement(11475..11505)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(11520..11541)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(11829..12417)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(12591..13448)
/label="AmpR"
/note="beta-lactamase"
promoter complement(13449..13553)
/label="AmpR promoter"