pLVTHM

$193.00

  • Model: PVT11115
  • 50 Units in Stock
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pLVTHM


Catalog No. PVT11115
Packing 2ug

 

pLVTHM Information

Promoter: EF-1 alpha, H1, SV40

Replicator: pUC ori, SV40 ori

Terminator: SV40 poly (A) signal, bGH poly (A) signal

Plasmid classification: virus series, lentivirus interference carrier

Plasmid size: 11085bp

Prokaryotic resistance: Amp

Cloned strain: Stbl3

Culture conditions: 37 centigrade, aerobic LB

Expression host: mammalian cells

Induction mode: no induction, instantaneous expression

5'sequencing primers: primers designed according to sequence

3'sequencing primers: WPRE-R:CATAGCGTAAAAGGAGCAACA

 

pLVTHM Description

pLVTHM allows for direct cloning of shRNAs into the lentiviral vectors and replaces the old pLVTH system from Trono lab. pLVTHM is similar to pLVTH, but contains a 3bp substitution that generates a unique MluI site for direct cloning of an shRNA into MluI-ClaI. Please note that there is an additional ClaI site in this vector that is blocked by Dam methylation . The plasmid needs to be grown in a Dam+ bacterial strain in order to use ClaI for cloning. Note that ClaI has lower salt concentration requirements than MluI. One way to handle this is to digest with ClaI for 45 minutes, followed by addition of diluted MluI buffer and MluI, then incubation for 90 more minutes. Be sure not to use a large amount of vector (1.7 ug to 2.5 ug is known to work). Also, if your shRNA is already in pSUPER (or another plasmid under the control of the PolIII promoter) you may use the EcoRI-ClaI sites to replace the H1 promoter in pLVTHM with the H1-shRNA cassette from pSUPER (or another plasmid).

pLVTHM-vector

pLVTHM Sequence

LOCUS       Exported               11085 bp ds-DNA     circular SYN 09-SEP-2016
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled 3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 11085)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, September 9, 2016 from SnapGene Viewer 3.1.4
             
FEATURES             Location/Qualifiers
     source          1..11085
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    168..302
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     CDS             complement(727..747)
                     /codon_start=1
                     /product="nuclear localization signal of SV40 large T 
                     antigen"
                     /note="SV40 NLS"
                     /translation="PKKKRKV"
     intron          877..942
                     /note="small t intron"
                     /note="simian virus 40 (SV40) small t antigen intron"
     promoter        complement(2076..2405)
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      2090..2225
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     rep_origin      complement(2651..3239)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3410..4270)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(4271..4375)
                     /gene="bla"
                     /note="AmpR promoter"
     promoter        5163..5181
                     /note="SP6 promoter"
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     LTR             5206..5386
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     protein_bind    complement(5406..5439)
                     /bound_moiety="Cre recombinase"
                     /note="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        complement(5474..5688)
                     /note="H1 promoter"
                     /note="human H1 RNA promoter"
     protein_bind    5717..5987
                     /bound_moiety="tetracycline repressor TetR"
                     /note="tetracycline response element"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     misc_feature    6155..6743
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             6261..6272
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /note="Factor Xa site"
                     /translation="IEGR"
     CDS             complement(6816..7532)
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    7622..7739
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        complement(7787..8964)
                     /note="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
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