lentiCas9-EGFP vector (V012330)

Basic Vector Information

      • Vector Name:
      • lentiCas9-EGFP
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 13177 bp
      • Type:
      • CRISPR Plasmids
      • Source/Author:
      • Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song
      • Copy Number:
      • High copy number

lentiCas9-EGFP vector Vector Map

lentiCas9-EGFP13177 bp6001200180024003000360042004800540060006600720078008400900096001020010800114001200012600CMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTSEF-1-alpha core promoterCas9nucleoplasmin NLSFLAGP2AEGFPWPRE5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

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 ℃.

lentiCas9-EGFP vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       lentiCas9-EGFP.       13177 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Zhang lab lentiviral vector for expressing human codon-optimized S. 
            pyogenes Cas9 plus enhanced GFP (EGFP).
ACCESSION   .
VERSION     .
KEYWORDS    lentiCas9-EGFP.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 13177)
  AUTHORS   Chen S, Sanjana NE, Zheng K, Shalem O, Lee K, Shi X, Scott DA, Song 
            J, Pan JQ, Weissleder R, Lee H, Zhang F, Sharp PA.
  TITLE     Genome-wide CRISPR screen in a mouse model of tumor growth and 
            metastasis.
  JOURNAL   Cell 2015;160:1246-60.
  PUBMED    25748654
REFERENCE   2  (bases 1 to 13177)
  AUTHORS   
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 13177)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell"; 
            date: "2015"; volume: "160"; pages: "1246-60"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..13177
                     /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        2650..2861
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             2886..6989
                     /label=Cas9
                     /note="Cas9 (Csn1) endonuclease from the Streptococcus
                     pyogenes Type II CRISPR/Cas system"
     CDS             6990..7037
                     /codon_start=1
                     /product="bipartite nuclear localization signal from
                     nucleoplasmin"
                     /label=bipartite nuclear localization signal from
                     nucl
                     /note="nucleoplasmin NLS"
                     /translation="KRPAATKKAGQAKKKK"
     CDS             7038..7061
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             7071..7127
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1
                     polyprotein"
                     /label=2A peptide from porcine teschovirus-1
                     polyprotein
                     /note="P2A"
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             7128..7841
                     /label=EGFP
                     /note="enhanced GFP"
     misc_feature    7869..8457
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8982..9162
                     /label=5' LTR (truncated)
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    9194..9418
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9464..9892
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        10017..10227
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        10283..10330
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             10349..10720
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    10853..10986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(11071..11101)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(11116..11137)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(11425..12013)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(12187..13044)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(13045..13149)
                     /label=AmpR promoter

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