TRMPV-ns vector (V001587)

Basic Vector Information

      • Vector Name:
      • TRMPV-ns
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 7838 bp
      • Type:
      • Retroviral Tet-shRNA expression vector
      • Source/Author:
      • Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe SW.

TRMPV-ns vector Vector Map

TRMPV-ns7838 bp300600900120015001800210024002700300033003600390042004500480051005400570060006300660069007200750078005'-LTRMMLV Psigag (truncated)TRE promoterdsRedmiR30-shRNAPGKVenus3' LTR (Delta-U3)SV40 promoteroriAmpRAmpR promoter

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 ℃.

TRMPV-ns vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                7838 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Retroviral Tet-shRNA expression vector TRMPV-ns, complete sequence.
ACCESSION   HQ456316
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7838)
  AUTHORS   Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe 
            SW.
  TITLE     Toolkit for evaluating genes required for proliferation and survival
            using tetracycline-regulated RNAi
  JOURNAL   Nat. Biotechnol. (2010) In press
  PUBMED    21131983
REFERENCE   2  (bases 1 to 7838)
  AUTHORS   Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe 
            SW.
  TITLE     Direct Submission
  JOURNAL   Submitted (01-NOV-2010) Scott Lowe Lab, Cold Spring Harbor 
            Laboratory, 1 Bungtown Road, Cold Spring Harbor, NY 11724, USA
REFERENCE   3  (bases 1 to 7838)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7838
                     /organism="Retroviral Tet-shRNA expression vector
                     TRMPV-ns"
                     /mol_type="other DNA"
                     /db_xref="taxon:931126"
     repeat_region   1..728
                     /label=5'-LTR
                     /note="5'-LTR"
     promoter        310..513
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     LTR             514..689
                     /label=5' LTR (truncated)
                     /note="truncated long terminal repeat from Moloney murine 
                     sarcoma virus"
     misc_feature    752..951
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     regulatory      758..1567
                     /regulatory_class="other"
                     /note="packaging signal"
     CDS             1152..1568
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     regulatory      1602..2041
                     /regulatory_class="promoter"
                     /note="TRE promoter"
     protein_bind    1618..1888
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        1921..1959
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             2058..2735
                     /codon_start=1
                     /gene="dsRed"
                     /product="dsRed"
                     /label=dsRed
                     /protein_id="ADR71680.1"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     gene            2058..2735
                     /gene="dsRed"
                     /label=dsRed
     misc_RNA        2777..3124
                     /product="miR30-shRNA"
                     /label=miR30-shRNA shRen.713 targeting Renilla luciferase
                     /note="miR30-shRNA shRen.713 targeting Renilla luciferase"
     ncRNA           2804..2925
                     /label=5' miR-30a
                     /note="sequence upstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2953..2967
                     /label=miR-30a loop
                     /note="loop from the the 71-nt precursor of the human 
                     miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     ncRNA           2995..3121
                     /label=3' miR-30a
                     /note="sequence downstream of the 71-nt precursor of the 
                     human miR-30a microRNA (Zeng et al., 2002)"
                     /ncRNA_class="miRNA"
     regulatory      3135..3643
                     /regulatory_class="promoter"
                     /note="PGK"
     promoter        3144..3643
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3666..4385
                     /codon_start=1
                     /gene="Venus"
                     /product="Venus"
                     /label=Venus
                     /protein_id="ADR71681.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            3666..4385
                     /gene="Venus"
                     /label=Venus
     LTR             4544..4969
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     promoter        5235..5564
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5415..5550
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(5857..6445)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6616..7476)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="ADR71682.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(6616..7476)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(7477..7581)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        7767..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"

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