Basic Vector Information
- Vector Name:
- pMK1200-Stuffer-EF1a-Puro-mCherry
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8626 bp
- Type:
- Transcriptional regulation
- Replication origin:
- ori
- Host:
- Mammalian cells, Lentivirus
- Selection Marker:
- Puro
- Promoter:
- EF-1α
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
pMK1200-Stuffer-EF1a-Puro-mCherry 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 ℃.
pMK1200-Stuffer-EF1a-Puro-mCherry vector Sequence
LOCUS 62056_17010 8626 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 8626)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..8626
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 247..626
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 627..829
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
LTR 844..1024
/label=5' LTR (truncated)
/note="truncated 5' long terminal repeat (LTR) from HIV-1"
misc_feature 1071..1196
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1695..1928
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 2113..2157
/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 2306..2347
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2455..2572
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2662..3834
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3846..4442
/label=PuroR
/note="puromycin N-acetyltransferase"
CDS 4518..5225
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
misc_feature 5515..6103
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
primer_bind complement(6106..6122)
/label=KS primer
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(6874..7462)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7636..8493)
/label=AmpR
/note="beta-lactamase"
promoter complement(8494..8598)
/label=AmpR promoter
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