pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro vector (V014242)

Price Information

Cat No. Plasmid Name Availability Add to cart
V014242 pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro In stock (lyophilized plasmid)

Buy one, get one free!

Two vials of lyophilized plasmid will be delivered, each vial is about 5µg.

Basic Vector Information

The pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro is for dual expression of Cas9 and sgRNA. It also contains a mCherry marker (red) and puromycin resistance.

Vector Name:
pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro
Antibiotic Resistance:
Ampicillin
Length:
13278 bp
Type:
Gene knockout
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Puro
Promoter:
U6
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro vector Vector Map

pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro13278 bp600120018002400300036004200480054006000660072007800840090009600102001080011400120001260013200U6 promotergRNA scaffoldEF-1-alpha core promoterCas9FLAGmCherryWPRE3' LTR (Delta-U3)bGH poly(A) signalf1 oriSV40 promoterPuroRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoterCMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTS

Plasmid Resuspension 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 ℃.

pLV2-U6-MCS-sgRNA-Cas9-mCherry-Puro vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V014242                13278 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014242
VERSION     V014242
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 13278)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..13278
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..241
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        266..341
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        403..614
                     /label="EF-1-alpha core promoter"
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     CDS             639..4742
                     /label="Cas9"
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             4791..4814
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             4881..5588
                     /label="mCherry"
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     misc_feature    5607..6195
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6267..6500
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6532..6756
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6802..7230
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        7244..7573
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     CDS             7621..8214
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     polyA_signal    8347..8480
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(8565..8595)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(8610..8631)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(8919..9507)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9681..10538)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(10539..10643)
                     /label="AmpR promoter"
     enhancer        10909..11288
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        11289..11491
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             11506..11686
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    11733..11858
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    12351..12584
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             12769..12813
                     /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             12962..13003
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    13111..13228
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"