Basic Vector Information
- Vector Name:
- pMLLV-EnCMV-MCS-3×FLAG-PGK-Puro
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8165 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- mPGK
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pMLLV-EnCMV-MCS-3×FLAG-PGK-Puro 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 ℃.
pMLLV-EnCMV-MCS-3×FLAG-PGK-Puro vector Sequence
LOCUS 62056_17155 8165 bp DNA circular SYN 01-JAN-1980
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8165)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8165
/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"
misc_feature 2802..2867
/label=MCS
/note="multiple cloning site"
CDS 2868..2933
/label=3xFLAG
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
promoter 2953..3452
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 3473..4069
/label=PuroR
/note="puromycin N-acetyltransferase"
misc_feature 4086..4674
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 4881..5514
/label=5' LTR
/note="5' long terminal repeat (LTR) from HIV-1"
primer_bind complement(5643..5659)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(5667..5683)
/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(5691..5721)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(5736..5757)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(6045..6633)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6807..7664)
/label=AmpR
/note="beta-lactamase"
promoter complement(7665..7769)
/label=AmpR promoter
polyA_signal 7817..7951
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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