pTet-O-Ngn2-puro vector (V006881)

Basic Vector Information

      • Vector Name:
      • pTet-O-Ngn2-puro
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9886 bp
      • Type:
      • Mammalian Expression, Lentiviral
      • Selection Marker:
      • Puromycin
      • Copy Number:
      • High Copy
      • Promoter:
      • tet-on
      • Cloning Method:
      • Restriction Enzyme
      • 5' Primer:
      • LNCX
      • 3' Primer:
      • WPRE-R

pTet-O-Ngn2-puro vector Vector Map

pTet-O-Ngn2-puro9886 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092009600CMV enhancerCMV promoter5' LTR (truncated)HIV-1 PsiRREgp41 peptideProtein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509cPPT/CTStetracycline response elementminimal CMV promoterKozak sequenceFLAGNeurog2T2APuroRWPREKS primer5' LTR (truncated)bGH poly(A) signalf1 oriSV40 promoterEM7 promoterBleoRSV40 poly(A) signalM13 revlac operatorlac promoterCAP binding siteL4440oriAmpRAmpR promoterpRS-marker

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 ℃.

pTet-O-Ngn2-puro vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pTet-O-Ngn2-puro        9886 bp DNA     circular SYN 13-MAY-2021
DEFINITION  3rd generation lentiviral vector, expresses Ngn2 and puromycin 
            resistent gene under control of TetON promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pTet-O-Ngn2-puro.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9886)
  AUTHORS   Zhang Y, Pak C, Han Y, Ahlenius H, Zhang Z, Chanda S, Marro S, 
            Patzke C, Acuna C, Covy J, Xu W, Yang N, Danko T, Chen L, Wernig M, 
            Sudhof TC
  TITLE     Rapid single-step induction of functional neurons from human 
            pluripotent stem cells.
  JOURNAL   Neuron. 2013 Jun 5;78(5):785-98. doi: 10.1016/j.neuron.2013.05.029.
  PUBMED    23764284
REFERENCE   2  (bases 1 to 9886)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 9886)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1016/j.neuron.2013.05"; journalName: "Neuron"; date: "2013-06-5-
            5"; volume: "78"; issue: "5"; pages: "785-98"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9886
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        100..479
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        480..682
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             697..877
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    924..1049
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1542..1775
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1960..2004
                     /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             2153..2194
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2302..2419
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    2476..2746
                     /label=tetracycline response element
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2779..2817
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     primer_bind     2814..2838
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      2917..2926
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2926..2949
                     /label=FLAG
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     CDS             2950..3735
                     /gene="Neurog2"
                     /label=Neurog2
                     /note="Neurogenin-2 from Mus musculus. Accession#: P70447"
     CDS             3742..3795
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             3802..4398
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    4440..5028
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     primer_bind     complement(5031..5047)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             5553..5733
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     polyA_signal    5765..5989
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6035..6463
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6477..6806
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     promoter        6854..6901
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             6920..7291
                     /label=BleoR
                     /note="antibiotic-binding protein"
     polyA_signal    7424..7557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7594..7610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7618..7634)
                     /label=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)."
     promoter        complement(7642..7672)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7687..7708)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     primer_bind     complement(7825..7842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7996..8584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8758..9615)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9616..9720)
                     /label=AmpR promoter
     primer_bind     complement(9795..9814)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"

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