Basic Vector Information
FUW-M2rtTA vector Vector Map
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 ℃.
FUW-M2rtTA vector Sequence
LOCUS FUW-M2rtTA. 7979 bp DNA circular SYN 13-MAY-2021 DEFINITION Lentiviral plasmid expressing the reverse tetracycline transactivator. ACCESSION . VERSION . KEYWORDS FUW-M2rtTA. SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7979) AUTHORS Hockemeyer D, Soldner F, Cook EG, Gao Q, Mitalipova M, Jaenisch R TITLE A drug-inducible system for direct reprogramming of human somatic cells to pluripotency. JOURNAL Cell Stem Cell. 2008 Sep 11. 3(3):346-53. PUBMED 18786421 REFERENCE 2 (bases 1 to 7979) TITLE Direct Submission REFERENCE 3 (bases 1 to 7979) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Stem Cell. 2008 Sep 11. 3(3):346-53." COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..7979 /mol_type="other DNA" /organism="synthetic DNA construct" primer_bind complement(46..65) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 237..616 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 617..819 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" LTR 834..1014 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" misc_feature 1061..1186 /label=HIV-1 Psi /note="packaging signal of human immunodeficiency virus type 1" misc_feature 1679..1912 /label=RRE /note="The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm." CDS 2097..2141 /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 2290..2331 /note="Protein Tat from Human immunodeficiency virus type 1 group M subtype B (isolate WMJ22). Accession#: P12509" misc_feature 2439..2556 /label=cPPT/CTS /note="central polypurine tract and central termination sequence of HIV-1" promoter 2621..3832 /label=UbC promoter /note="human ubiquitin C promoter" CDS 3861..4604 /label=rtTA-Advanced /note="improved tetracycline-controlled transactivator" misc_feature 4644..5232 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(5235..5251) /label=KS primer /note="common sequencing primer, one of multiple similar variants" LTR 5757..5937 /label=5' LTR (truncated) /note="truncated 5' long terminal repeat (LTR) from HIV-1" polyA_signal 5969..6080 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin complement(6226..6814) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(6988..7845) /label=AmpR /note="beta-lactamase" promoter complement(7846..7950) /label=AmpR promoter
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