Basic Vector Information
- Vector Name:
- pNBioSec
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7032 bp
- Type:
- Mammalian expression vector
- Replication origin:
- ori
- Source/Author:
- Mize GJ, Harris JE, Takayama TK, Kulman JD.
- Promoter:
- SV40
pNBioSec 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 ℃.
pNBioSec vector Sequence
LOCUS 40924_33053 7032 bp DNA circular SYN 18-DEC-2018
DEFINITION Mammalian expression vector pNBioSec, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7032)
AUTHORS Mize GJ, Harris JE, Takayama TK, Kulman JD.
TITLE Regulated expression of active biotinylated G-protein coupled
receptors in mammalian cells
JOURNAL Protein Expr. Purif. 57 (2), 280-289 (2008)
PUBMED 18042400
REFERENCE 2 (bases 1 to 7032)
AUTHORS Mize GJ, Harris JE, Takayama TK, Kulman JD.
TITLE Direct Submission
JOURNAL Submitted (04-AUG-2007) Department of Biochemistry, University of
Washington, N.E. Pacific Street, Seattle, WA 98195, USA
REFERENCE 3 (bases 1 to 7032)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7032)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "2008"; volume: "57"; issue: "2"; pages:
"280-289"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(04-AUG-2007) Department of Biochemistry, University of Washington,
N.E. Pacific Street, Seattle, WA 98195, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7032
/mol_type="other DNA"
/organism="synthetic DNA construct"
protein_bind 15..285
/label=tetracycline response element
/note="contains seven copies of the tetracycline operator
tetO"
promoter 318..356
/label=minimal CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
regulatory 477..485
/label=Kozak site
/note="Kozak site"
/regulatory_class="ribosome_binding_site"
misc_feature 570..608
/label=biotin acceptor peptide (BioTag)-encoding
/note="biotin acceptor peptide (BioTag)-encoding"
misc_feature 609..628
/note="multiple cloning site; MCS"
misc_feature 729..1315
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
regulatory 1326..1334
/label=Kozak site
/note="Kozak site"
/regulatory_class="ribosome_binding_site"
CDS 1335..1424
/label=prolactin signal sequence
/note="signal sequence from bovine prolactin"
CDS 1425..1448
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 1455..2417
/label=BirA
/note="E. coli biotin protein ligase"
intron 2485..3057
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
CDS 3061..3183
/gene="HBB"
/label=HBB
/note="Hemoglobin subunit beta from Colobus guereza.
Accession#: Q9TT33"
polyA_signal 3254..3309
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(3871..4459)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4633..5490)
/label=AmpR
/note="beta-lactamase"
promoter complement(5491..5595)
/label=AmpR promoter
promoter 5700..6029
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 6122..6718
/label=PuroR
/note="puromycin N-acetyltransferase"
polyA_signal 6951..7032
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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