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 Map

pUB_smFLAG_H2B_MS29371 bp4008001200160020002400280032003600400044004800520056006000640068007200760080008400880092003' LTRHIV-1 PsiRREgp41 peptideProtein TatcPPT/CTSUbC promoterFLAGFLAGFLAGFLAGFLAGFLAGFLAGFLAGFLAGFLAGH2BWPRE3' LTR (Delta-U3)pBRforEcoAmpR promoterAmpRoriL4440SV40pro-FCAP binding sitelac promoterlac operatorM13 revEBV-rev

pUB_smFLAG_H2B_MS2 vector Sequence

Copy Sequence

Download GenBank 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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