Basic Vector Information
- Vector Name:
- 3XFlagNICD1
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7057 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- hGH-pA-R
3XFlagNICD1 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 ℃.
3XFlagNICD1 vector Sequence
LOCUS 40924_85 7057 bp DNA circular SYN 13-MAY-2021
DEFINITION synthetic circular DNA.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7057)
AUTHORS Ong CT, Cheng HT, Chang LW, Ohtsuka T, Kageyama R, Stormo GD, Kopan
R
TITLE Target selectivity of vertebrate notch proteins. Collaboration
between discrete domains and CSL-binding site architecture
determines activation probability.
JOURNAL J Biol Chem. 2006 Feb 24. 281(8):5106-19.
PUBMED 16365048
REFERENCE 2 (bases 1 to 7057)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7057)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J Biol
Chem. 2006 Feb 24. 281(8):5106-19."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7057
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 107..128
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 143..173
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 181..197
/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)."
primer_bind 205..221
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 235..253
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
promoter complement(292..621)
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
polyA_signal complement(650..1272)
/label=hGH poly(A) signal
/note="human growth hormone polyadenylation signal"
CDS complement(3674..3739)
/codon_start=1
/label=3xFLAG
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
/translation="DYKDHDGDYKDHDIDYKDDDDK"
promoter complement(3769..3972)
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
enhancer complement(3973..4352)
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
primer_bind complement(4513..4529)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(4670..5125)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 5152..5256
/label=AmpR promoter
CDS 5257..6114
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
rep_origin 6288..6876
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 7030..7047
/label=L4440
/note="L4440 vector, forward primer"
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