Basic Vector Information
- Vector Name:
- pML-EnCMV-FLAG-Ub-K6
- Antibiotic Resistance:
- Ampicillin
- Length:
- 3973 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Promoter:
- CMV
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pML-EnCMV-FLAG-Ub-K6 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 ℃.
pML-EnCMV-FLAG-Ub-K6 vector Sequence
LOCUS 62056_17095 3973 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 3973)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..3973
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 27..330
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 331..534
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
intron 672..768
/label=SV40 intron
/note="modified SV40 intron with splice donor and acceptor
sites"
CDS 832..855
/codon_start=1
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/translation="DYKDDDDK"
CDS 859..1086
/codon_start=1
/label=ubiquitin-K6
/note="human ubiquitin variant containing Lys-6, with all
other lysines mutated to arginine"
/translation="MQIFVKTLTGRTITLEVEPSDTIENVRARIQDREGIPPDQQRLIF
AGRQLEDGRTLSDYNIQRESTLHLVLRLRGG"
polyA_signal complement(1112..1246)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(1357..1373)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind complement(1381..1397)
/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(1405..1435)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(1450..1471)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(1759..2347)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(2521..3378)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
promoter complement(3379..3483)
/label=AmpR promoter
primer_bind 3957..3973
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
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