Basic Vector Information
- Vector Name:
- pTgKCNQ2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 14164 bp
- Type:
- Targeting vector
- Replication origin:
- ori
- Source/Author:
- Tomonoh Y, Deshimaru M, Araki K, Miyazaki Y, Arasaki T, Tanaka Y, Kitamura H, Mori F, Wakabayashi K, Yamashita S, Saito R, Itoh M, Uchida T, Yamada J, Migita K, Ueno S, Kitaura H, Kakita A, Lossin C,
pTgKCNQ2 vector Map
pTgKCNQ2 vector Sequence
LOCUS 40924_43153 14164 bp DNA circular SYN 18-DEC-2018
DEFINITION Targeting vector pTgKCNQ2 DNA, complete sequence.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 14164)
AUTHORS Tomonoh Y, Deshimaru M, Araki K, Miyazaki Y, Arasaki T, Tanaka Y,
Kitamura H, Mori F, Wakabayashi K, Yamashita S, Saito R, Itoh M,
Uchida T, Yamada J, Migita K, Ueno S, Kitaura H, Kakita A, Lossin C,
Takano Y, Hirose S.
TITLE The kick-in system: a novel rapid knock-in strategy
JOURNAL PLoS ONE 9 (2), E88549 (2014)
PUBMED 24586341
REFERENCE 2 (bases 1 to 14164)
AUTHORS Deshimaru M, Miyazaki Y, Arasaki T, Araki K, Hirose S.
TITLE Direct Submission
JOURNAL Submitted (24-NOV-2009) Contact:Masanobu Deshimaru Fukuoka
University, Department of Chemistry, Faculty of Science; Nanakuma
8-19-1, Jonan-ku, Fukuoka, Fukuoka 814-0180, Japan
REFERENCE 3 (bases 1 to 14164)
TITLE Direct Submission
REFERENCE 4 (bases 1 to 14164)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE";
date: "2014"; volume: "9"; issue: "2"; pages: "E88549"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(24-NOV-2009) Contact:Masanobu Deshimaru Fukuoka University,
Department of Chemistry, Faculty of Science; Nanakuma 8-19-1,
Jonan-ku, Fukuoka, Fukuoka 814-0180, Japan"
COMMENT SGRef: number: 3; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..14164
/mol_type="other DNA"
/organism="synthetic DNA construct"
rep_origin complement(3..458)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind 600..616
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
promoter 626..644
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 961..1533
/label=DTA
/note="diphtheria toxin fragment A"
intron 1610..1675
/label=small t intron
/note="SV40 (simian virus 40) small t antigen intron"
polyA_signal 1888..1943
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
misc_feature 2316..7483
/gene="Kcnq2"
/standard_name="long arm"
/function="long arm for homologous recombination"
/label=long arm
/note="Kcnq2 sequence stretching from intron 1 to intron 5,
which induce recombination at the homologous genomic region
in mouse ES cell"
protein_bind complement(7484..7517)
/label=lox71
/note="Left element (LE) mutant of loxP (Araki et al.,
2010). Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
misc_feature 7518..8090
/gene="Kcnq2"
/note="Kcnq2 genomic segment consisting of exon 6 and
flanking intronic sequence, which can be exchanged with the
mutated segment via Cre-lox mediated specific
recombination."
protein_bind complement(8093..8126)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
protein_bind complement(8183..8230)
/label=FRT
/bound_moiety="FLP recombinase from the Saccharomyces
cerevisiae 2u plasmid"
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
promoter 8301..8801
/label=PGK promoter
/note="mouse phosphoglycerate kinase 1 promoter"
CDS 8815..9615
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase from Tn5"
protein_bind complement(9669..9702)
/label=lox2272
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are
compatible with each other, but incompatible with loxP or
loxN sites (Lee and Saito, 1988)."
protein_bind complement(10041..10088)
/label=FRT
/note="FLP-mediated recombination occurs in the 8-bp core
sequence TCTAGAAA (Turan and Bode, 2011)."
misc_feature 10145..11939
/gene="Kcnq2"
/standard_name="short arm"
/function="short arm for homologous recombination"
/label=short arm
/note="Kcnq2 sequence stretching from intron 6 to intron 8,
which induces recombination at the homologous genomic
region in mouse ES cell"
gene 10145..11939
/gene="Kcnq2"
/label=Kcnq2
promoter complement(11976..11994)
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
primer_bind complement(12015..12031)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 12039..12055
/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)."
promoter complement(12063..12093)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(12108..12129)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(12417..13005)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(13179..14036)
/label=AmpR
/note="beta-lactamase"
promoter complement(14037..14141)
/label=AmpR promoter
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