Basic Vector Information
- Vector Name:
- FBDki
- Antibiotic Resistance:
- Ampicillin
- Length:
- 10504 bp
- Type:
- Knock-in vector
- Replication origin:
- ori
- Source/Author:
- Tamayev R, Giliberto L, Matsuda S, D'Abramo C, Arancio O, Vidal R, D'Adamio L.
- Promoter:
- mPGK
FBDki 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 ℃.
FBDki vector Sequence
LOCUS 40924_810 10504 bp DNA circular SYN 17-DEC-2018
DEFINITION Knock-in vector FBDki, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 10504)
AUTHORS Tamayev R, Giliberto L, Matsuda S, D'Abramo C, Arancio O, Vidal R,
D'Adamio L.
TITLE Memory deficits due to Familial British Dementia BRI2 mutation are
caused by loss of BRI2 function rather than amyloidosis
JOURNAL Unpublished
REFERENCE 2 (bases 1 to 10504)
AUTHORS Giliberto L, D'Adamio L.
TITLE Direct Submission
JOURNAL Submitted (24-MAY-2010) Litwin-Zucker Center for the Study of
Alzheimer's Disease, The Feinstein Institute for Medical
Research-North Shore-LIJ, 350 Community Drive, Manhasset, NY 11030,
USA
REFERENCE 3 (bases 1 to 10504)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 10504)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Unpublished"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-MAY-2010) Litwin-Zucker Center for the Study of Alzheimer's
Disease, The Feinstein Institute for Medical Research-North
Shore-LIJ, 350 Community Drive, Manhasset, NY 11030, USA"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..10504
/mol_type="other DNA"
/organism="synthetic DNA construct"
promoter 24..128
/label=AmpR promoter
CDS 129..986
/label=AmpR
/note="beta-lactamase"
rep_origin 1160..1748
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
protein_bind 2036..2057
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 2072..2102
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 2110..2126
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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 2134..2150
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 2171..2189
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
polyA_signal 2257..2481
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
intron complement(2555..2620)
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
CDS complement(2695..3267)
/label=DTA
/note="diphtheria toxin fragment A"
regulatory complement(3286..3797)
/label=PGK1 promoter
/note="PGK1 promoter"
/regulatory_class="promoter"
promoter complement(3294..3788)
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
misc_feature 3828..4989
/note="left arm; contains intron 5"
misc_feature 5038..5072
/label=LoxP
/note="LoxP"
protein_bind 5038..5071
/label=loxP
/bound_moiety="Cre recombinase"
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (GCATACAT)."
polyA_signal complement(5130..5354)
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
CDS complement(5395..6195)
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
promoter complement(6209..6703)
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
protein_bind complement(6733..6766)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
primer_bind complement(6780..6796)
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 6805..6825
/label=pBlueScript MCS fragment
/note="pBlueScript MCS fragment"
primer_bind 6805..6821
/label=SK primer
/note="common sequencing primer, one of multiple similar
variants"
misc_feature 6829..9835
/note="right arm fragment; contains intron 5, exon 6, and
intron 6"
exon 7037..7156
/number=6
CDS 7037..7156
/codon_start=1
/product="ITM2B integral membrane protein 2B"
/label=ITM2B integral membrane protein 2B
/note="ABRI"
/protein_id="ADO40090.1"
/translation="GIQKREASNCFAIRHFENKFAVETLICSRTVKKNIIEEN"
misc_difference 7070
/replace="a"
/label=humanized
/note="humanized"
misc_feature 7121..7156
/label=ABRI mutant-extra bases
/note="ABRI mutant-extra bases"
misc_difference 7121
/replace="t"
/label=ABRI mutation
/note="ABRI mutation"
3'UTR 7157..7960
/label=ITM2B 3' UTR
/note="ITM2B 3' UTR"
promoter complement(9861..9879)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(9889..9905)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin 10047..10502
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
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