Price Information
| Cat No. | Plasmid Name | Availability | Add to cart |
|---|---|---|---|
| V012345 | pXPR_120 | 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_120
- Antibiotic Resistance:
- Ampicillin
- Length:
- 15513 bp
- Type:
- Mammalian Expression, Lentiviral
- Replication origin:
- ori
- Selection Marker:
- Blasticidin
- Promoter:
- EF-1α
pXPR_120 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.
pXPR_120 vector Sequence
LOCUS V012345 15513 bp DNA circular SYN 09-JUN-2021
DEFINITION Exported.
ACCESSION V012345
VERSION V012345
KEYWORDS pXPR_120
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 15513)
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 15513)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 15513)
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..15513
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(30..49)
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
primer_bind 276..296
/label="pBABE 3'"
/note="SV40 enhancer, reverse primer for pBABE vectors"
rep_origin complement(305..733)
/direction=LEFT
/label="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
polyA_signal complement(779..1003)
/label="bGH poly(A) signal"
/note="bovine growth hormone polyadenylation signal"
LTR complement(1035..1215)
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
primer_bind 1720..1736
/label="pBluescriptKS"
/note="For pBluescript vector"
primer_bind 1721..1737
/label="KS primer"
/note="common sequencing primer, one of multiple similar
variants"
misc_feature complement(1740..2328)
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
CDS complement(2356..2751)
/label="BSD"
/note="blasticidin S deaminase"
CDS complement(2752..2805)
/label="T2A"
/note="2A peptide from Thosea asigna virus capsid protein"
CDS complement(2821..3390)
/label="Rta AD"
/note="transcriptional activation domain from the human
herpesvirus 4 (Epstein-Barr virus) replication and
transcription activator Rta/BRLF1 (Hardwick et al., 1992;
Chavez et al., 2015)"
CDS complement(3409..3765)
/label="RelA (p65) AD"
/note="transcriptional activation domain of human RelA,
also known as p65 (O'Shea and Perkins, 2008)"
CDS complement(4195..4215)
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS complement(4240..4389)
/label="VP64"
/note="tetrameric repeat of the minimal activation domain
of herpes simplex virus VP16 (Beerli et al., 1998)"
CDS complement(4429..4449)
/label="SV40 NLS"
/note="nuclear localization signal of SV40 (simian virus
40) large T antigen"
CDS complement(4504..8604)
/label="dCas9"
/note="catalytically dead mutant of the Cas9 endonuclease
from the Streptococcus pyogenes Type II CRISPR/Cas system"
CDS complement(8605..8652)
/codon_start=1
/product="bipartite nuclear localization signal from
nucleoplasmin"
/label="nucleoplasmin NLS"
/translation="KRPAATKKAGQAKKKK"
promoter complement(8668..9846)
/label="EF-1-alpha promoter"
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature complement(9976..10093)
/label="cPPT/CTS"
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(10201..10242)
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
/label="Protein Tat"
CDS complement(10391..10435)
/label="gp41 peptide"
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
misc_feature complement(10620..10853)
/label="RRE"
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
misc_feature complement(11346..11471)
/label="HIV-1 Psi"
/note="packaging signal of human immunodeficiency virus
type 1"
LTR complement(11518..11698)
/label="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
promoter complement(11713..11915)
/label="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(11916..12295)
/label="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
primer_bind 12467..12486
/label="pRS-marker"
/note="pRS vectors, use to sequence yeast selectable
marker"
promoter 12561..12665
/label="AmpR promoter"
CDS 12666..13523
/label="AmpR"
/note="beta-lactamase"
rep_origin 13697..14285
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 14439..14456
/label="L4440"
/note="L4440 vector, forward primer"
protein_bind 14573..14594
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 14609..14639
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 14647..14663
/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)."
primer_bind 14671..14687
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
polyA_signal complement(14724..14857)
/label="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
CDS complement(14990..15361)
/label="BleoR"
/note="antibiotic-binding protein"
promoter complement(15380..15427)
/label="EM7 promoter"
/note="synthetic bacterial promoter"