pHAGE2-TetOminiCMV-SKM vector (V016805)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016805 pHAGE2-TetOminiCMV-SKM In stock, 1 week for quality controls

Buy one, get one free!

Two tubes of lyophilized plasmid will be delivered, each tube is about 5µg.

Basic Vector Information

Vector Name:
pHAGE2-TetOminiCMV-SKM
Antibiotic Resistance:
Ampicillin
Length:
10186 bp
Type:
Protein expression
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Promoter:
Tet
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pHAGE2-TetOminiCMV-SKM vector Map

pHAGE2-TetOminiCMV-SKM10186 bp50010001500200025003000350040004500500055006000650070007500800085009000950010000mSox2mKlf4mMycWPRE5' LTR (truncated)AmpR promoterAmpRoriCAP binding sitelac promoter3' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTStet operatortet operator

Plasmid 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 ℃.

6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it

General Plasmid Transform Protocol

1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.

2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.

3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.

4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.

5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.

pHAGE2-TetOminiCMV-SKM vector Sequence

LOCUS       V016805                10186 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016805
VERSION     V016805
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10186)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10186
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             173..1129
                     /label="mSox2"
                     /note="Mus musculus transcription factor SOX-2 gene.
                     Belongs to the SRY-related HMG-box (SOX) family of
                     transcription factors, which is involved in the regulation
                     of embryonic development and in the determination of cell
                     fate"
     CDS             1205..2653
                     /label="mKlf4"
                     /note="Mouse Kruppel-like factor (Klf4) gene. Belongs to
                     the relatively large family of SP1-like transcription
                     factors and is involved in the regulation of proliferation,
                     differentiation, apoptosis and somatic cell reprogramming."
     CDS             2738..4099
                     /label="mMyc"
                     /note="Mus musculus myc proto-oncogene. Belongs to
                     myelocytomatosis (Myc) family of transcription factors.
                     Plays a role in cell cycle procession, apotosis and
                     cellular transformation"
     misc_feature    4116..4704
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             4852..5032
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     promoter        5165..5269
                     /label="AmpR promoter"
     CDS             5270..6127
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      6301..6889
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    7300..7321
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7336..7366
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     LTR             7675..8308
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    8355..8480
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    8977..9210
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             9395..9439
                     /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             9588..9629
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    9732..9849
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    9925..9943
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    10009..10027
                     /label="tet operator"
                     /note="bacterial operator O2 for the tetR and tetA genes"