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
Replication origin:
ori
Source/Author:
Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena
Copy Number:
High copy number
Promoter:
EF-1α

lenti sgRNA(MS2)_zeo backbone vector Map

lenti sgRNA(MS2)_zeo backbone9996 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600CMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSU6 promoterMS2 stem loopMS2 stem looppolIII terminatorEF-1-alpha promoterBleoRWPRE5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoter

lenti sgRNA(MS2)_zeo backbone vector Sequence

LOCUS       V012332                 9996 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V012332
VERSION     V012332
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"
            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"
                     /label="Protein Tat"
     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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