pLVX-PAmCherry-C1 vector (V012717)

MSDS

Basic Vector Information

pLVX-PAmCherry-C1 Vector is an HIV-1-based, lentiviral expression vector. Lentiviral particles derived from the vector allow you to infect cells and express your gene of interest fused to PAmCherry, a photoactivatable mutant of the fluorescent protein mCherry. PAmCherry is non-fluorescence until photoactivated by a short exposure to light at a wavelength between 350 nm and 400 nm. The excitation/emission wavelengths of photoactivated PAmCherry are 564 nm and 595 nm. Genes cloned into the MCS will be expressed as fusions to the C-terminus of PAmCherry if they are in the same reading frame as PAmCherry and there are no intervening stop codons.

Vector Name:: pLVX-PAmCherry-C1

Antibiotic Resistance:: Ampicillin

Length:: 8770 bp

Type:: Viral Expression & Packaging Vectors

Replication origin:: ori

Source/Author:: Clontech

Selection Marker:: Puromycin

Copy Number:: High copy number

Promoter:: mPGK

Cloning Method:: Enzyme Cut

Fusion Tag:: PAmCherry

Expression Method:: Constiutive, Stable

pLVX-PAmCherry-C1 vector Vector Map

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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 ℃.

pLVX-PAmCherry-C1 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

LOCUS       pLVX-PAmCherry-C        8770 bp DNA     circular SYN 08-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pLVX-PAmCherry-C1.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8770)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 8770)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..8770
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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             1914..1955
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2027..2144
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        2201..2504
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2505..2708
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             2806..3513
                     /label=PAmCherry1
                     /note="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
     misc_feature    3517..3582
                     /label=MCS
                     /note="multiple cloning site"
     promoter        3605..4104
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             4125..4721
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    4738..5326
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             5533..6166
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     primer_bind     complement(6295..6311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6319..6335)
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(6343..6373)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6388..6409)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6697..7285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7459..8316)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(8317..8421)
                     /label=AmpR promoter
     polyA_signal    8469..8603
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"

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