Tet-pLKO-neo vector (V012736)

Basic Vector Information

3rd generation lentiviral plasmid for inducible expression of shRNA; neomycin selection.

Vector Name:
Tet-pLKO-neo
Antibiotic Resistance:
Ampicillin
Length:
10959 bp
Type:
RNAi
Replication origin:
ori
Source/Author:
Dmitri Wiederschain
Selection Marker:
Neomycin/G418(Geneticin)
Copy Number:
High copy number
Promoter:
hPGK

Tet-pLKO-neo vector Map

Tet-pLKO-neo10959 bp5001000150020002500300035004000450050005500600065007000750080008500900095001000010500AmpR promoterAmpRoriCAP binding sitelac promoterlac operatorM13 revT3 promoterRSV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein TatH1-2O2 promotercPPT/CTShPGK promoterbeta-globin intronT7 promoterTetRIRESNeoR/KanRattL23' LTR (Delta-U3)SV40 poly(A) signalSV40 oriT7 promoterM13 fwdM13 ori

Tet-pLKO-neo vector Sequence

LOCUS       V012736                10959 bp    DNA     circular SYN 11-SEP-2021
DEFINITION  Exported.
ACCESSION   V012736
VERSION     V012736
KEYWORDS    Tet-pLKO-neo
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10959)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 10959)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10959
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        106..210
                     /label="AmpR promoter"
     CDS             211..1068
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1242..1830
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2118..2139
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2154..2184
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2192..2208
                     /label="lac repressor encoded by lacI binding site"
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2216..2232
                     /label="M13 rev"
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2253..2271
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2299..2525
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2526..2706
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2753..2878
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    3371..3604
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3789..3833
                     /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             3982..4023
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     promoter        4122..4216
                     /label="H1-2O2 promoter"
                     /note="doxycycline-inducible variant of the human H1 RNA
                     promoter (Henriksen et al., 2007)"
     misc_feature    6134..6251
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        6300..6810
                     /label="hPGK promoter"
                     /note="human phosphoglycerate kinase 1 promoter"
     intron          6819..7391
                     /label="beta-globin intron"
                     /note="intron from rabbit beta-globin gene"
     promoter        7446..7464
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             7475..8095
                     /label="TetR"
                     /note="tetracycline repressor TetR"
     misc_feature    8130..8703
                     /label="IRES"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             8710..9501
                     /label="NeoR/KanR"
                     /note="aminoglycoside phosphotransferase"
     protein_bind    complement(9533..9632)
                     /label="attL2"
                     /note="recombination site for the Gateway(R) LR reaction"
     LTR             9776..10009
                     /label="3' LTR (Delta-U3)"
                     /note="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    10081..10215
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      10242..10377
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(10398..10416)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(10426..10442)
                     /label="M13 fwd"
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      10584..10959
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"

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