pUB_smFLAG_H2B_MS2 vector (V012827)

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 Vector Map

pUB_smFLAG_H2B_MS29371 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092003' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSUbC promotervertebrate consensus sequence for strong initiation of translation (Kozak, 1987)FLAGFLAGFLAGFLAGFLAGFLAGFLAGFLAGFLAGFLAGH2BWPRE3' LTR (Delta-U3)pBRforEcoAmpR promoterAmpRoriL4440SV40pro-FCAP binding sitelac promoterlac operatorM13 revEBV-rev

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_H2B_MS2 vector Sequence

Copy Sequence

Download GeneBank File(.gb)

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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