Basic Vector Information
BASU RaPID plasmid 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 ℃.
BASU RaPID plasmid vector Sequence
LOCUS BASU_RaPID_plasm 12356 bp DNA circular SYN 13-MAY-2021
DEFINITION Expresses the BASU RaPID labeling protein.
ACCESSION .
VERSION .
KEYWORDS BASU RaPID plasmid.
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 12356)
AUTHORS Ramanathan M, Majzoub K, Rao DS, Neela PH, Zarnegar BJ, Mondal S,
Roth JG, Gai H, Kovalski JR, Siprashvili Z, Palmer TD, Carette JE,
Khavari PA
TITLE RNA-protein interaction detection in living cells.
JOURNAL Nat Methods. 2018 Feb 5. pii: nmeth.4601. doi: 10.1038/nmeth.4601.
PUBMED 29400715
REFERENCE 2 (bases 1 to 12356)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 12356)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2018 Feb 5. pii: nmeth.4601. doi: 10.1038/nmeth.4601."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..12356
/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 2063..2180
/label=cPPT/CTS
/note="central polypurine tract and central termination
sequence of HIV-1"
CDS complement(2248..2619)
/label=BleoR
/note="antibiotic-binding protein"
promoter complement(2639..2686)
/label=EM7 promoter
/note="synthetic bacterial promoter"
enhancer 2791..3094
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 3095..3298
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 3295..3319
/label=LNCX
/note="Human CMV promoter, forward primer"
CDS 3492..3518
/label=NES
/note="nuclear export signal from the HIV Rev protein
(Fischer et al., 1995)"
CDS 3528..3554
/label=HA
/note="HA (human influenza hemagglutinin) epitope tag"
misc_feature 4358..4940
/label=IRES2
/note="internal ribosome entry site (IRES) of the
encephalomyocarditis virus (EMCV)"
CDS 4939..5532
/label=PuroR
/note="puromycin N-acetyltransferase"
CDS 5542..5595
/codon_start=1
/product="2A peptide from Thosea asigna virus capsid
protein"
/label=T2A
/note="Eukaryotic ribosomes fail to insert a peptide bond
between the Gly and Pro residues, yielding separate
polypeptides."
/translation="EGRGSLLTCGDVEENPGP"
CDS 5608..6315
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
misc_feature 6331..6919
/label=WPRE
/note="woodchuck hepatitis virus posttranscriptional
regulatory element"
LTR 7126..7359
/label=3' LTR (Delta-U3)
/note="self-inactivating 3' long terminal repeat (LTR) from
HIV-1"
polyA_signal 7466..7690
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 7736..8164
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 8178..8507
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 8556..9578
/label=HygR
/note="aminoglycoside phosphotransferase from E. coli"
polyA_signal 9711..9844
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(9881..9897)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(9881..9897)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(9894..9916)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 9905..9921
/label=lac operator
/bound_moiety="lac repressor encoded by lacI"
/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)."
promoter complement(9929..9959)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind complement(9974..9995)
/label=CAP binding site
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(10112..10129)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(10283..10871)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(11045..11902)
/label=AmpR
/note="beta-lactamase"
promoter complement(11903..12007)
/label=AmpR promoter
polyA_signal 12055..12189
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
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