Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V007241 | pXPR_123 | 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:
- pXPR_123
- Antibiotic Resistance:
- Ampicillin
- Length:
- 15807 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Blasticidin
- Promoter:
- EF-1α
pXPR_123 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.
pXPR_123 vector Sequence
LOCUS V007241 15807 bp DNA circular SYN 13-MAY-2021
DEFINITION Exported.
ACCESSION V007241
VERSION V007241
KEYWORDS pXPR_123
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 15807)
AUTHORS Najm FJ, Strand C, Donovan KF, Hegde M, Sanson KR, Vaimberg EW,
Sullender ME, Hartenian E, Kalani Z, Fusi N, Listgarten J, Younger
ST, Bernstein BE, Root DE, Doench JG
TITLE Orthologous CRISPR-Cas9 enzymes for combinatorial genetic screens.
JOURNAL Nat Biotechnol. 2018 Feb;36(2):179-189. doi: 10.1038/nbt.4048. Epub
2017 Dec 18.
PUBMED 29251726
REFERENCE 2 (bases 1 to 15807)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 15807)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10.1038/nbt.4048";
journalName: "Nat Biotechnol"; date: "2018-02"; volume: "36"; issue:
"2"; pages: "179-189"
SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..15807
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 248..676
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 690..1019
/label="SV40 promoter"
/note="SV40 enhancer and early promoter"
promoter 1067..1114
/label="EM7 promoter"
/note="synthetic bacterial promoter"
CDS 1133..1504
/label="BleoR"
/note="antibiotic-binding protein"
polyA_signal 1637..1770
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
primer_bind complement(1807..1823)
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1831..1847)
/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(1855..1885)
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind complement(1900..1921)
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(2038..2055)
/label="L4440"
/note="L4440 vector, forward primer"
rep_origin complement(2209..2797)
/direction=LEFT
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2971..3828)
/label="AmpR"
/note="beta-lactamase"
promoter complement(3829..3933)
/label="AmpR promoter"
primer_bind complement(4008..4027)
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 4199..4578
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 4579..4781
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 4796..4976
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 5023..5148
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 5641..5874
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 6059..6103
/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 6252..6293
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
misc_feature 6401..6518
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 6648..7826
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 7842..7889
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label="nucleoplasmin NLS"
/translation="KRPAATKKAGQAKKKK"
CDS 7890..11990
/label="dCas9"
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS 12045..12065
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS 12081..13931
/label="p300 core"
/note="catalytic histone acetyltransferase core of the
human E1A-associated protein p300 (Delvecchio et al.,
2013)"
CDS 13932..13958
/label="HA"
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 13983..14036
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS 14037..14432
/label="BSD"
/note="blasticidin S deaminase"
misc_feature 14460..15048
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(15051..15067)
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(15052..15068)
/label="pBluescriptKS"
/note="For pBluescript vector"
LTR 15573..15753
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(15779..15796)
/label="BGH-rev"
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"