lentiGuide-Puro vector (V012327)

Price Information

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V012327 lentiGuide-Puro In stock (lyophilized plasmid)

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

Basic Vector Information

The lentiGuide-Puro is a lentiviral vector, also known as pLentiGuide-Puro. It is for expressing a single guide RNA (sgRNA) in Cas9-expressing cells.

      • Vector Name:
      • lentiGuide-Puro
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 10183 bp
      • Type:
      • CRISPR Plasmids
      • Source/Author:
      • Sanjana NE, Shalem O, Zhang F.
      • Copy Number:
      • High copy number
      • Growth Strain(s):
      • Stbl3
      • Growth Temperature:
      • 37℃

lentiGuide-Puro vector Vector Map

lentiGuide-Puro10183 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000EF-1-alpha promoterPuroRWPRE3' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7 promoterf1 oriAmpR promoterAmpRoriCAP binding sitelac promoterT3 promoterRSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideU6 promoterfillergRNA scaffoldpolIII terminatorcPPT/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 ℃.

References

  • Datlinger P, Rendeiro AF, Schmidl C, Krausgruber T, Traxler P, Klughammer J, Schuster LC, Kuchler A, Alpar D, Bock C. Pooled CRISPR screening with single-cell transcriptome readout. Nat Methods. 2017 Mar;14(3):297-301.

lentiGuide-Puro vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       lentiGuide-Puro.       10183 bp DNA     circular SYN 01-JAN-1980
DEFINITION  Zhang lab lentiviral vector, also known as pLentiGuide-Puro, for 
            expressing a single guide RNA (sgRNA) in Cas9-expressing cells. Use 
            together with lentiCas9-Blast.
ACCESSION   .
VERSION     .
KEYWORDS    lentiGuide-Puro.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10183)
  AUTHORS   Sanjana NE, Shalem O, Zhang F.
  TITLE     Improved vectors and genome-wide libraries for CRISPR screening.
  JOURNAL   Nat. Methods 2014;11:783-4.
  PUBMED    25075903
REFERENCE   2  (bases 1 to 10183)
  AUTHORS   Zhang Lab / Addgene #52963
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 10183)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat. 
            Methods"; date: "2014"; volume: "11"; pages: "783-4"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10183
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        87..1265
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1278..1874
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    1893..2481
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             2553..2786
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    2864..2998
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      3025..3160
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        complement(3181..3199)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     rep_origin      3367..3822
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3848..3952
                     /label=AmpR promoter
     CDS             3953..4810
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      4984..5572
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    5860..5881
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        5896..5926
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     promoter        5995..6013
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        6041..6267
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             6268..6448
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    6495..6620
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    7113..7346
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             7531..7575
                     /label=gp41 peptide
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et 
                     al., 2013)"
     promoter        7753..7993
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_feature    7998..9882
                     /label=filler
                     /note="filler"
                     /note="digest with BsmBI to replace the filler with an
                     sgRNA"
     misc_RNA        9883..9958
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      9959..9964
                     /note="polIII terminator"
                     /note="RNA polymerase III transcription terminator"
     misc_feature    10014..10131
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"