TtRMPVIR vector (V001585)

Basic Vector Information

      • Vector Name:
      • TtRMPVIR
      • Antibiotic Resistance:
      • Ampicillin
      • Length:
      • 9040 bp
      • Type:
      • Retroviral Tet-shRNA expression vector
      • Source/Author:
      • Zuber J, McJunkin K, Fellmann C, Dow LE, Taylor MJ, Hannon GJ, Lowe SW.

TtRMPVIR vector Vector Map

TtRMPVIR9040 bp400800120016002000240028003200360040004400480052005600600064006800720076008000840088005'-LTRMMLV Psigag (truncated)TREtightdsRedmiR30-shRNAPGKVenusIRESrtTA33' 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 ℃.

TtRMPVIR vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       Exported                9040 bp ds-DNA     circular SYN 18-DEC-2018
DEFINITION  Retroviral Tet-shRNA expression vector TtRMPVIR, complete sequence.
ACCESSION   HQ456318
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9040)
  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 9040)
  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 9040)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9040
                     /organism="Retroviral Tet-shRNA expression vector
                     TtRMPVIR"
                     /mol_type="other DNA"
                     /db_xref="taxon:931128"
     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      1599..1914
                     /regulatory_class="promoter"
                     /note="TREtight"
     promoter        1600..1914
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    1608..1626
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1644..1662
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1679..1697
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1715..1733
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1751..1769
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1786..1804
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1822..1840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     CDS             1953..2630
                     /codon_start=1
                     /gene="dsRed"
                     /product="dsRed"
                     /label=dsRed
                     /protein_id="ADR71687.1"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     gene            1953..2630
                     /gene="dsRed"
                     /label=dsRed
     misc_RNA        2644..2991
                     /product="miR30-shRNA"
                     /label=miR30-shRNA shRen.713 targeting Renilla luciferase
                     /note="miR30-shRNA shRen.713 targeting Renilla luciferase"
     ncRNA           2671..2792
                     /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           2820..2834
                     /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           2862..2988
                     /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      3002..3510
                     /regulatory_class="promoter"
                     /note="PGK"
     promoter        3011..3510
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3533..4252
                     /codon_start=1
                     /gene="Venus"
                     /product="Venus"
                     /label=Venus
                     /protein_id="ADR71688.1"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     gene            3533..4252
                     /gene="Venus"
                     /label=Venus
     misc_feature    4282..4833
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             4845..5591
                     /codon_start=1
                     /gene="rtTA3"
                     /product="rtTA3"
                     /label=rtTA3
                     /protein_id="ADR71689.1"
                     /translation="MSRLDKSKVINGALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHFCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     gene            4845..5591
                     /gene="rtTA3"
                     /label=rtTA3
     LTR             5746..6171
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     Moloney murine leukemia virus"
     promoter        6437..6766
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6617..6752
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     rep_origin      complement(7059..7647)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7818..8678)
                     /codon_start=1
                     /gene="ampR"
                     /product="AmpR"
                     /label=ampR
                     /protein_id="ADR71690.1"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     gene            complement(7818..8678)
                     /gene="ampR"
                     /label=ampR
     promoter        complement(8679..8783)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        8969..309
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"

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