Basic Vector Information
- Vector Name:
- pNBio
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6895 bp
- Type:
- Mammalian expression vector
- Replication origin:
- ori
- Source/Author:
- Kulman JD, Satake M, Harris JE.
- Promoter:
- SV40
pNBio 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 ℃.
pNBio vector Sequence
LOCUS 40924_33048 6895 bp DNA circular SYN 18-DEC-2018
DEFINITION Mammalian expression vector pNBio, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6895)
AUTHORS Kulman JD, Satake M, Harris JE.
TITLE A versatile system for site-specific enzymatic biotinylation and
regulated expression of proteins in cultured mammalian cells
JOURNAL Protein Expr. Purif. 52 (2), 320-328 (2007)
PUBMED 17084093
REFERENCE 2 (bases 1 to 6895)
AUTHORS Kulman JD, Satake M, Harris JE.
TITLE Direct Submission
JOURNAL Submitted (28-APR-2006) Biochemistry, University of Washington, N.E.
Pacific St., Seattle, WA 98195, USA
REFERENCE 3 (bases 1 to 6895)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 6895)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Protein
Expr. Purif."; date: "2007"; volume: "52"; issue: "2"; pages:
"320-328"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(28-APR-2006) Biochemistry, University of Washington, N.E. Pacific
St., Seattle, WA 98195, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6895
/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"
regulatory 480..489
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
misc_feature 492..530
/label=biotin acceptor peptide (BioTag)-encoding
/note="biotin acceptor peptide (BioTag)-encoding"
misc_feature 532..551
/note="multiple cloning site; MCS"
misc_feature 652..1238
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
regulatory 1270..1278
/gene="birA"
/label=Kozak site
/note="Kozak site"
/regulatory_class="ribosome_binding_site"
regulatory 1273..1282
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 1282..1305
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 1318..2280
/label=BirA
/note="E. coli biotin protein ligase"
intron 2348..2920
/label=beta-globin intron
/note="intron from rabbit beta-globin gene"
CDS 2924..3046
/gene="HBB"
/label=HBB
/note="Hemoglobin subunit beta from Colobus guereza.
Accession#: Q9TT33"
polyA_signal 3117..3172
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
rep_origin complement(3734..4322)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(4496..5353)
/label=AmpR
/note="beta-lactamase"
promoter complement(5354..5458)
/label=AmpR promoter
promoter 5563..5892
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 5985..6581
/label=PuroR
/note="puromycin N-acetyltransferase"
polyA_signal 6814..6895
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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