Basic Vector Information
p28BIOH-LIC4 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 ℃.
p28BIOH-LIC4 vector Sequence
LOCUS 40924_2508 7373 bp DNA circular SYN 18-DEC-2018
DEFINITION Expression vector p28BIOH-LIC4, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7373)
AUTHORS Loppnau PA, Graslund S, Arrowsmith CH.
TITLE A pipeline to generate recombinant antibodies to human target
proteins involved in epigenetic mechanisms
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 7373)
AUTHORS Loppnau PA, Graslund S, Arrowsmith CH.
TITLE Direct Submission
JOURNAL Submitted (13-NOV-2012) Structural Genomics Consortium, University
of Toronto, MaRS Centre, South tower, 101 College St, Room 700,
Toronto, Ontario M5G 1L7, Canada
REFERENCE 3 (bases 1 to 7373)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 7373)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(13-NOV-2012) Structural Genomics Consortium, University of Toronto,
MaRS Centre, South tower, 101 College St, Room 700, Toronto, Ontario
M5G 1L7, Canada"
COMMENT SGRef: number: 3; type: "Journal Article"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7373
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin 12..467
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
CDS complement(563..1375)
/label=KanR
/note="aminoglycoside phosphotransferase"
rep_origin 1497..2085
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
misc_feature complement(2271..2413)
/label=bom
/note="basis of mobility region from pBR322"
CDS complement(2518..2706)
/label=rop
/note="Rop protein, which maintains plasmids at low copy
number"
protein_bind complement(3481..3502)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
CDS complement(3518..4597)
/label=lacI
/note="lac repressor"
promoter complement(4598..4675)
/label=lacI promoter
promoter 4984..5002
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 5003..5027
/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)."
RBS 5042..5064
/label=RBS
/note="efficient ribosome binding site from bacteriophage
T7 gene 10 (Olins and Rangwala, 1989)"
CDS 5077..5121
/label=AviTag(TM)
/note="peptide tag that allows for enzymatic biotinylation"
misc_feature 5128..5142
/note="5'-In-Fusion sequence; add to upstream PCR primer"
protein_bind complement(5170..5203)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
promoter 5218..5663
/label=sacB promoter
/note="sacB promoter and control region"
CDS 5664..7082
/label=SacB
/note="secreted levansucrase that renders bacterial growth
sensitive to sucrose"
misc_feature 7158..7172
/note="3'-In-Fusion sequence; add complement to downstream
PCR primer"
CDS 7170..7187
/label=6xHis
/note="6xHis affinity tag"
CDS 7217..7234
/label=6xHis
/note="6xHis affinity tag"
terminator 7301..7348
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
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