pINDUCER11(miR-RUG) vector (V016793)

Price Information

Cat No. Plasmid Name Availability Add to cart
V016793 pINDUCER11(miR-RUG) 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:
pINDUCER11(miR-RUG)
Antibiotic Resistance:
Ampicillin
Length:
14701 bp
Type:
RNA interference, Tetracycline inducible
Replication origin:
ori
Host:
Mammalian cells, Lentivirus
Promoter:
UbC
Growth Strain(s):
Stbl3
Growth Temperature:
37℃

pINDUCER11(miR-RUG) vector Vector Map

pINDUCER11(miR-RUG)14701 bp700140021002800350042004900560063007000770084009100980010500112001190012600133001400014700RREgp41 peptideProtein TatcPPT/CTSBleoREM7 promotertetracycline response elementminimal CMV promoterTurboRFP5' miR-30athrombin siteHRV 3C sitePhenylalanine--tRNA ligase alpha subunitTEV site3' miR-30aUbC promoterloxPrtTA-AdvancedIRESEGFPWPRE3' LTR (Delta-U3)bGH poly(A) signalM13 oriSV40 promoterSV40 poly(A) signallac promoterCAP binding siteoriAmpRAmpR promoterSV40 poly(A) signal3' LTRHIV-1 Psi

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.

pINDUCER11(miR-RUG) vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       V016793                14701 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V016793
VERSION     V016793
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 14701)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..14701
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    500..733
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             918..962
                     /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             1111..1152
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    1260..1377
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     CDS             complement(1445..1816)
                     /label="BleoR"
                     /note="antibiotic-binding protein"
     promoter        complement(1836..1883)
                     /label="EM7 promoter"
                     /note="synthetic bacterial promoter"
     protein_bind    1953..2223
                     /label="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator
                     tetO"
     promoter        2256..2294
                     /label="minimal CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2391..3083
                     /label="TurboRFP"
                     /note="red fluorescent protein from Entacmaea quadricolor"
     ncRNA           3137..3230
                     /label="5' miR-30a"
                     /note="sequence upstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     CDS             3268..3285
                     /label="thrombin site"
                     /note="thrombin recognition and cleavage site"
     CDS             3286..3309
                     /label="HRV 3C site"
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases"
     CDS             complement(3486..4466)
                     /gene="pheS"
                     /label="Phenylalanine--tRNA ligase alpha subunit"
                     /note="Phenylalanine--tRNA ligase alpha subunit from
                     Escherichia coli O139:H28 (strain E24377A / ETEC).
                     Accession#: A7ZMI2"
     CDS             4604..4624
                     /label="TEV site"
                     /note="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
     ncRNA           4648..4760
                     /label="3' miR-30a"
                     /note="sequence downstream of the 71-nt precursor of the
                     human miR-30a microRNA (Zeng et al., 2002)"
     promoter        4811..6022
                     /label="UbC promoter"
                     /note="human ubiquitin C promoter"
     protein_bind    6029..6062
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             6069..6812
                     /label="rtTA-Advanced"
                     /note="improved tetracycline-controlled transactivator"
     misc_feature    6864..7436
                     /label="IRES"
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
     CDS             7454..8170
                     /label="EGFP"
                     /note="enhanced GFP"
     misc_feature    8195..8783
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8990..9223
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    9330..9554
                     /label="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      9600..9980
                     /label="M13 ori"
                     /note="M13 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        10042..10371
                     /label="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     polyA_signal    11194..11275
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     promoter        complement(11412..11442)
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(11457..11478)
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(11766..12354)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(12528..13385)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(13386..13490)
                     /label="AmpR promoter"
     polyA_signal    13538..13672
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     LTR             13840..14473
                     /label="3' LTR"
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    14520..14645
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"