Basic Vector Information
For expression of a circular nLuc mRNA. To express your gene of interest clone into the BsiWI, SacII sites and add the GCCATGTcTATGTGG 5' of the SacII site.
- Vector Name:
- pcDNA3.1+-Tornado-nLuc
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7008 bp
- Type:
- Mammalian Expression Vectors
- Source/Author:
- Samie Jaffrey Lab
- Selection Marker:
- Neomycin/G418(Geneticin)
- Copy Number:
- High copy number
- Promoter:
- CMV
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
pcDNA3.1+-Tornado-nLuc vector Map
pcDNA3.1+-Tornado-nLuc vector Sequence
LOCUS Exported 7008 bp DNA circular SYN 01-DEC-2025
DEFINITION For expression of a circular nLuc mRNA. To express your gene of
interest clone into the BsiWI, SacII sites and add the
GCCATGTcTATGTGG 5' of the SacII site..
ACCESSION .
VERSION .
KEYWORDS pcDNA3.1+-Tornado-nLuc
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7008)
AUTHORS Unti MJ, Jaffrey SR
TITLE Highly efficient cellular expression of circular mRNA enables
prolonged protein expression.
JOURNAL Cell Chem Biol. 2023 Oct 13:S2451-9456(23)00334-3. doi:
10.1016/j.chembiol.2023.09.015.
PUBMED 37883972
REFERENCE 2 (bases 1 to 7008)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Cell Chem
Biol. 2023 Oct 13:S2451-9456(23)00334-3. doi:
10.1016/j.chembiol.2023.09.015."
COMMENT Sequence Label: pcDNA3.1+-Tornado-nLuc
FEATURES Location/Qualifiers
source 1..7008
/mol_type="other DNA"
/organism="synthetic DNA construct"
source 1067..1081
/mol_type="other DNA"
/organism="synthetic DNA construct"
primer_bind complement(44..63)
/label=pRS-marker
/note="pRS vectors, use to sequence yeast selectable
marker"
enhancer 235..614
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 615..818
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 863..881
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 901..924
/codon_start=1
/product="FLAG(R) epitope tag, followed by an enterokinase
cleavage site"
/label=FLAG
/translation="DYKDDDDK"
CDS 937..963
/codon_start=1
/product="HA (human influenza hemagglutinin) epitope tag"
/label=HA
/translation="YPYDVPDYA"
promoter 988..1006
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
misc_feature 1007..1060
/label=Ribozyme
misc_feature 1061..1089
/label=StemForming
misc_feature 1092..1890
/label=IRES
CDS 1888..1890
/codon_start=1
/label=START
/translation="M"
CDS 1897..2409
/codon_start=1
/product="NanoLuc(R) luciferase"
/label=Nluc
/note="human codon-optimized"
/translation="VFTLEDFVGDWRQTAGYNLDQVLEQGGVSSLFQNLGVSVTPIQRI
VLSGENGLKIDIHVIIPYEGLSGDQMGQIEKIFKVVYPVDDHHFKVILHYGTLVIDGVT
PNMIDYFGRPYEGIAVFDGKKITVTGTLWNGNKIIDERLINPDGSLLFRVTINGVTGWR
LCERILA"
misc_RNA 2410..2428
/label=MS2 stem loop
/note="stem loop that binds the bacteriophage MS2 coat
protein"
misc_feature 2435..2471
/label=StemForming
misc_feature 2472..2525
/label=Twister Ribozyme
promoter complement(2561..2579)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
primer_bind complement(2562..2579)
/label=SP6
/note="SP6 promoter, forward primer"
primer_bind complement(2599..2616)
/label=BGH-rev
/note="Bovine growth hormone terminator, reverse primer.
Also called BGH reverse"
polyA_signal 2605..2829
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2875..3303
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
primer_bind complement(2962..2981)
/label=F1ori-R
/note="F1 origin, reverse primer"
primer_bind 3172..3193
/label=F1ori-F
/note="F1 origin, forward primer"
primer_bind complement(3312..3332)
/label=pBABE 3'
/note="SV40 enhancer, reverse primer for pBABE vectors"
promoter 3317..3646
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
rep_origin 3497..3632
/label=SV40 ori
/note="SV40 origin of replication"
primer_bind 3559..3578
/label=SV40pro-F
/note="SV40 promoter/origin, forward primer"
CDS 3713..4507
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/label=NeoR/KanR
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(3767..3786)
/label=Neo-R
/note="Neomycin resistance gene, reverse primer"
primer_bind 4377..4396
/label=Neo-F
/note="Neomycin resistance gene, forward primer"
polyA_signal 4681..4802
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4718..4737)
/label=SV40pA-R
/note="SV40 polyA, reverse primer"
primer_bind 4772..4791
/label=EBV-rev
/note="SV40 polyA terminator, reverse primer"
primer_bind complement(4851..4867)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
primer_bind complement(4851..4867)
/label=M13 Reverse
/note="In lacZ gene. Also called M13-rev"
primer_bind complement(4864..4886)
/label=M13/pUC Reverse
/note="In lacZ gene"
protein_bind 4875..4891
/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(4899..4929)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4944..4965
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
primer_bind complement(5082..5099)
/label=L4440
/note="L4440 vector, forward primer"
rep_origin complement(5253..5841)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
primer_bind complement(5333..5352)
/label=pBR322ori-F
/note="pBR322 origin, forward primer"
CDS complement(6012..6872)
/codon_start=1
/gene="bla"
/product="beta-lactamase"
/label=AmpR
/note="confers resistance to ampicillin, carbenicillin, and
related antibiotics"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
primer_bind 6635..6654
/label=Amp-R
/note="Ampicillin resistance gene, reverse primer"
promoter complement(6873..6977)
/gene="bla"
/label=AmpR promoter
This page is informational only.