Basic Vector Information
lenti sgRNA(MS2)_zeo backbone vector Vector Map
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 ℃.
lenti sgRNA(MS2)_zeo backbone vector Sequence
LOCUS lenti_sgRNA(MS2) 9996 bp DNA circular SYN 01-JAN-1980
DEFINITION Zhang lab lentiviral vector for expressing a Zeocin(TM)-resistance
marker plus a single guide RNA (sgRNA) containing two MS2 stem
loops.
ACCESSION .
VERSION .
KEYWORDS lenti sgRNA(MS2)_zeo backbone.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9996)
AUTHORS Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
C, Hsu PD, Habib N, Gootenberg JS, Nishimasu H, Nureki O, Zhang F.
TITLE Genome-scale transcriptional activation by an engineered CRISPR-Cas9
complex.
JOURNAL Nature 2015;517:583-8.
PUBMED 25494202
REFERENCE 2 (bases 1 to 9996)
AUTHORS Zhang Lab / Addgene #61427
TITLE Direct Submission
REFERENCE 3 (bases 1 to 9996)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nature";
date: "2015"; volume: "517"; pages: "583-8"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..9996
/mol_type="other DNA"
/organism="synthetic DNA construct"
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 Psi
/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
/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 2607..2847
/label=U6 promoter
/note="RNA polymerase III promoter for human U6 snRNA"
misc_RNA 2893..2911
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_RNA 2963..2981
/label=MS2 stem loop
/note="MS2 stem loop"
/note="stem loop that binds the bacteriophage MS2 coat
protein"
terminator 3012..3017
/note="polIII terminator"
/note="RNA polymerase III transcription terminator"
promoter 3105..4282
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 4289..4660
/label=BleoR
/note="antibiotic-binding protein"
misc_feature 4688..5276
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 5801..5981
/label=5' LTR (truncated)
/note="5' LTR (truncated)"
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
polyA_signal 6013..6237
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 6283..6711
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6725..7054
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 7102..7149
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 7168..7539
/label=BleoR
/note="antibiotic-binding protein"
polyA_signal 7672..7805
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
promoter complement(7890..7920)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(7935..7956)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(8244..8832)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(9006..9863)
/label=AmpR
/note="beta-lactamase"
promoter complement(9864..9968)
/label=AmpR promoter
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