Basic Vector Information
- Vector Name:
- Flag-HA-USP36
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9812 bp
- Type:
- Gene expression
- Replication origin:
- ori
- Host:
- Mammalian cells, Retrovirus
- Selection Marker:
- Puro
- Promoter:
- MSCV
- Growth Strain(s):
- Stbl3
- Growth Temperature:
- 37℃
Flag-HA-USP36 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 ℃.
Flag-HA-USP36 vector Sequence
LOCUS 62056_796 9812 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 9812)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..9812
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(234..822)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(996..1853)
/label=AmpR
/note="beta-lactamase"
promoter complement(1854..1958)
/label=AmpR promoter
LTR 2786..3302
/label=5' LTR
/note="5' long terminal repeat from murine embryonic stem
cell virus"
misc_feature 3366..3707
/label=MESV Psi
/note="packaging signal of murine embryonic stem cell
virus"
CDS 3774..4190
/label=gag (truncated)
/note="truncated Moloney murine leukemia virus (MMLV) gag
gene lacking the start codon"
CDS 4212..4235
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 4248..4274
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
CDS 4323..7691
/gene="USP36"
/label=USP36
/note="Ubiquitin carboxyl-terminal hydrolase 36 from Homo
sapiens. Accession#: Q9P275"
protein_bind complement(7702..7722)
/label=attB2
/note="core recombination site for the Gateway(R) BP
reaction"
misc_feature 7740..8316
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 8317..8913
/label=PuroR
/note="puromycin N-acetyltransferase"
LTR 8992..9506
/label=3' LTR
/note="3' long terminal repeat from murine embryonic stem
cell virus"
protein_bind complement(9668..9684)
/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(9692..9722)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9737..9758)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
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