Basic Vector Information
pUB_smFLAG_KDM5B_MS2 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 ℃.
pUB_smFLAG_KDM5B_MS2 vector Sequence
LOCUS pUB_smFLAG_KDM5B 13637 bp DNA circular SYN 13-MAY-2021
DEFINITION Translation reporter: Encodes KDM5B (1544 aa) fused to smFLAG
(Spaghetti Monster FLAG: 10 copies of an epitope tag FLAG). .
ACCESSION .
VERSION .
KEYWORDS pUB_smFLAG_KDM5B_MS2.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 13637)
AUTHORS Morisaki T, Lyon K, DeLuca KF, DeLuca JG, English BP, Zhang Z, Lavis
LD, Grimm JB, Viswanathan S, Looger LL, Lionnet T, Stasevich TJ
TITLE Real-time quantification of single RNA translation dynamics in
living cells.
JOURNAL Science. 2016 Jun 17;352(6292):1425-9. doi: 10.1126/science.aaf0899.
Epub 2016 May 5.
PUBMED 27313040
REFERENCE 2 (bases 1 to 13637)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 13637)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; doi:
"10.1126/science.aaf0899"; journalName: "Science"; date:
"2016-06-17- 17"; volume: "352"; issue: "6292"; pages: "1425-9"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..13637
/mol_type="other DNA"
/organism="synthetic DNA construct"
LTR 1..634
/label=3' LTR
/note="3' long terminal repeat (LTR) from HIV-1"
misc_feature 681..806
/label=HIV-1 Psi
/note="packaging signal of human immunodeficiency virus
type 1"
misc_feature 1303..1536
/label=RRE
/note="The Rev response element (RRE) of HIV-1 allows for
Rev-dependent mRNA export from the nucleus to the
cytoplasm."
CDS 1721..1765
/label=gp41 peptide
/note="antigenic peptide corresponding to amino acids 655
to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
al., 2013)"
CDS 1914..1955
/note="Protein Tat from Human immunodeficiency virus type 1
group M subtype B (isolate WMJ22). Accession#: P12509"
misc_feature 2058..2175
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
promoter 2218..2616
/label=UbC promoter
/note="human ubiquitin C promoter"
regulatory 2622..2631
/label=Kozak sequence
/note="vertebrate consensus sequence for strong initiation
of translation (Kozak, 1987)"
/regulatory_class="other"
CDS 2631..2654
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 2658..2681
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 2685..2708
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3213..3236
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3243..3266
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3279..3302
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3309..3332
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3525..3548
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3552..3575
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3579..3602
/label=FLAG
/note="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
CDS 3648..8279
/gene="KDM5B"
/label=KDM5B
/note="Lysine-specific demethylase 5B from Homo sapiens.
Accession#: Q9UGL1"
misc_feature 10027..10615
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 10690..10923
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(11042..11060)
/label=pBRforEco
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 11128..11232
/label=AmpR promoter
CDS 11233..12090
/label=AmpR
/note="beta-lactamase"
rep_origin 12264..12852
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 13006..13023
/label=L4440
/note="L4440 vector, forward primer"
primer_bind 13115..13134
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
protein_bind 13263..13284
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 13299..13329
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 13337..13353
/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 13361..13377
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 13427..13446
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
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