lenti sgRNA(MS2)_zeo backbone vector (V012332)

Basic Vector Information

      • Vector Name:
      • lenti sgRNA(MS2)_zeo backbone
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9996 bp
      • Type:
      • CRISPR Plasmids
      • Source/Author:
      • Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
      • Copy Number:
      • High copy number

lenti sgRNA(MS2)_zeo backbone vector Vector Map

lenti sgRNA(MS2)_zeo backbone9996 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600CMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREcPPT/CTSU6 promotergRNA scaffoldpolIII terminatorEF-1-alpha promoterBleoRWPRE5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signaloriAmpRAmpR 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 ℃.

lenti sgRNA(MS2)_zeo backbone vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9996 bp ds-DNA    circular SYN 06-1-2016
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
FEATURES             Location/Qualifiers
     source          1..9996
                     /organism="synthetic DNA construct"
                     /lab_host="Mammalian Cells"
                     /mol_type="other DNA"
     enhancer        238..617
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..817
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             835..1015
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1062..1187
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1680..1913
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2440..2557
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2607..2847
                     /note="U6 promoter"
                     /note="RNA polymerase III promoter for human U6 snRNA"
     misc_RNA        2876..3011
                     /note="gRNA scaffold"
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system, modified by the insertion of two MS2 
                     stem loops"
     misc_RNA        2893..2911
                     /note="MS2 stem loop"
                     /note="stem loop that binds the bacteriophage MS2 coat 
                     protein"
     misc_RNA        2963..2981
                     /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
                     /note="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          3336..4273
                     /note="EF-1-alpha intron A"
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     CDS             4289..4663
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="BleoR"
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     misc_feature    4688..5276
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     LTR             5801..5981
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    6013..6237
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6283..6711
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        6725..7054
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      6905..7040
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        7102..7149
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter "
     CDS             7168..7542
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /note="BleoR"
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    7672..7793
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(8244..8832)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9003..9863)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(9864..9968)
                     /gene="bla"
                     /note="AmpR promoter"

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