Basic Vector Information
Translation reporter: Encodes Histone H2B (125 aa) fused to smFLAG (Spaghetti Monster FLAG: 10 copies of an epitope tag FLAG).
- Vector Name:
- pUB_smFLAG_H2B_MS2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 9371 bp
- Type:
- Mammalian Expression Vectors
- Replication origin:
- ori
- Source/Author:
- Tim Stasevich
- Copy Number:
- High copy number
- Promoter:
- UbC
- Fusion Tag:
- Spaghetti Monster FLAG: 10 copies of an epitope tag FLAG
- Growth Strain(s):
- NEB Stable
- Growth Temperature:
- 37℃
pUB_smFLAG_H2B_MS2 vector Map
pUB_smFLAG_H2B_MS2 vector Sequence
LOCUS V012827 9371 bp DNA circular SYN 06-NOV-2023
DEFINITION Exported.
ACCESSION V012827
VERSION V012827
KEYWORDS pUB_smFLAG_H2B_MS2
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
.
REFERENCE 1 (bases 1 to 9371)
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 9371)
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"
FEATURES Location/Qualifiers
source 1..9371
/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"
/label="Protein Tat"
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
/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 3639..4013
/label="H2B"
/note="human histone H2B"
misc_feature 5761..6349
/label="WPRE"
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 6424..6657
/label="3' LTR (Delta-U3)"
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
primer_bind complement(6776..6794)
/label="pBRforEco"
/note="pBR322 vectors, upsteam of EcoRI site, forward
primer"
promoter 6862..6966
/label="AmpR promoter"
CDS 6967..7824
/label="AmpR"
/note="beta-lactamase"
rep_origin 7998..8586
/label="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind 8740..8757
/label="L4440"
/note="L4440 vector, forward primer"
primer_bind 8849..8868
/label="SV40pro-F"
/note="SV40 promoter/origin, forward primer"
protein_bind 8997..9018
/label="CAP binding site"
/note="CAP binding activates transcription in the presence
of cAMP."
promoter 9033..9063
/label="lac promoter"
/note="promoter for the E. coli lac operon"
protein_bind 9071..9087
/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 9095..9111
/label="M13 rev"
/note="common sequencing primer, one of multiple similar
variants"
primer_bind 9161..9180
/label="EBV-rev"
/note="SV40 polyA terminator, reverse primer"
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