pLentiV2-dCas9-VP64 vector (V017668)

Basic Vector Information

Vector Name:
pLentiV2-dCas9-VP64
Antibiotic Resistance:
Ampicillin
Length:
15499 bp
Type:
Gene-editing vector
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Selection Marker:
Puro
Promoter:
EF1a,U6
5' Primer:
G35310-F1:AGGTACCTGAGGTGTGACTG
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pLentiV2-dCas9-VP64 vector Map

pLentiV2-dCas9-VP6415499 bp70014002100280035004200490056006300700077008400910098001050011200119001260013300140001470015400CMV enhancerCMV promoter5' LTR (truncated)HIV-1 ΨRREgp41 peptideProtein TatcPPT/CTSU6 promotergRNA scaffoldEF-1α core promoterdCas9SV40 NLSVP64P2APuroRWPRE5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

pLentiV2-dCas9-VP64 vector Sequence

LOCUS       .                      15499 bp    DNA     circular UNK 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   
VERSION     
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        238..617
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        618..820
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             835..1015
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /label="HIV-1 Ψ"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1680..1913
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2098..2142
                     /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             2291..2332
                     /gene="Protein Tat"
                     /label="Protein Tat"
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2440..2557
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        2608..2848
                     /label="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        4738..4813
                     /label="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes
                     CRISPR/Cas9 system"
     promoter        4875..5086
                     /label="EF-1α core promoter"
                     /note="core promoter for human elongation factor EF-1α"
     CDS             5151..9251
                     /label="dCas9"
                     /note="catalytically dead mutant of the Cas9 endonuclease
                     from the Streptococcus pyogenes Type II CRISPR/Cas system"
     CDS             9306..9326
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             9351..9500
                     /label="VP64"
                     /note="tetrameric repeat of the minimal activation domain
                     of herpes simplex virus VP16 (Beerli et al., 1998)"
     CDS             9522..9578
                     /label="P2A"
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
     CDS             9579..10172
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     misc_feature    10191..10779
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             11304..11484
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    11516..11740
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      11786..12214
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        12228..12557
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     promoter        12605..12652
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     CDS             12671..13042
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     polyA_signal    13175..13308
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(13393..13423)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(13438..13459)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(13747..14335)
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(14509..15366)
                     /label="AmpR"
                     /note="β-lactamase"
     promoter        complement(15367..15471)
                     /label="AmpR promoter"

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