Basic Vector Information
RN3P-KDM5B-FLAG 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 ℃.
RN3P-KDM5B-FLAG vector Sequence
LOCUS RN3P-KDM5B-FLAG. 7915 bp DNA circular SYN 13-MAY-2021
DEFINITION FLAG-tagged mouse KDM5B cDNA clone for mammalian cell expression.
ACCESSION .
VERSION .
KEYWORDS RN3P-KDM5B-FLAG.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7915)
AUTHORS Zhang B, Zheng H, Huang B, Li W, Xiang Y, Peng X, Ming J, Wu X,
Zhang Y, Xu Q, Liu W, Kou X, Zhao Y, He W, Li C, Chen B, Li Y, Wang
Q, Ma J, Yin Q, Kee K, Meng A, Gao S, Xu F, Na J, Xie W
TITLE Allelic reprogramming of the histone modification H3K4me3 in early
mammalian development.
JOURNAL Nature. 2016 Sep 14;537(7621):553-557. doi: 10.1038/nature19361.
PUBMED 27626382
REFERENCE 2 (bases 1 to 7915)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 7915)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi: "10"; journalName:
"Nature"; date: "2016-09-14- 14"; volume: "537"; issue: "7621";
pages: "553-557"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..7915
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind 432..451
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
primer_bind 685..702
/label=L4440
/note="L4440 vector, forward primer"
protein_bind 819..840
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 855..885
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 893..909
/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)."
primer_bind 917..933
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
promoter 954..972
/label=T3 promoter
/note="promoter for bacteriophage T3 RNA polymerase"
5'UTR 997..1039
/label=Xenopus globin 5'-UTR
/note="translational enhancer from the 5'-UTR of the major
beta-globin gene of Xenopus laevis"
CDS 1069..1134
/label=3xFLAG
/note="three tandem FLAG(R) epitope tags, followed by an
enterokinase cleavage site"
CDS 1180..5811
/gene="Kdm5b"
/label=Kdm5b
/note="Lysine-specific demethylase 5B from Mus musculus.
Accession#: Q80Y84"
3'UTR 5841..5966
/label=Xenopus globin 3'-UTR
/note="3'-UTR of the major beta-globin gene of Xenopus
laevis"
promoter complement(6057..6075)
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
primer_bind complement(6082..6098)
/label=M13 fwd
/note="common sequencing primer, one of multiple similar
variants"
rep_origin complement(6240..6695)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 6722..6826
/label=AmpR promoter
CDS 6827..7684
/label=AmpR
/note="beta-lactamase"
rep_origin 7858..7915
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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