Human EEF1A1 ORF clone (NM_001402) (Cat. No.: V035085)
Note: A plasmid for high-efficiency expression of Myc-tagged EEF1A1 protein in mammalian cells
- Name:
- pcDNA3.1-EEF1A1(human)-Myc
- Accession ID:
- NM_001402.6
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6826 bp
- Type:
- Protein expression
- Replication origin:
- ori
- Host:
- Mammalian cells
- Selection Marker:
- Neo/G418
- Copy Number:
- High copy number
- Promoter:
- CMV
- 5' Primer:
- CMV-F
- Fusion Tag:
- Myc
- Growth Strain(s):
- DH5a
- Growth Temperature:
- 37℃
- Expression Method:
- Transient
- Gene Synonyms:
- CCS-3; CCS3; EE1A1; EEF-1; EEF1A; eEF1A-1; EF-Tu; EF1A; EF1A1; EF1alpha1; GRAF-1EF; LENG7; PTI1
- Transcript Definition:
- Homo sapiens eukaryotic translation elongation factor 1 alpha 1 (EEF1A1), mRNA
Resources
Plasmid Protocol
1. Centrifuge at 5,000×g for 5 min.
2. Carefully open the tube and add sterile water to dissolve the DNA: add 20 μl for 5 μg plasmid, and 100 μl for 100 μg plasmid.
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 ℃.
6. The concentration of plasmid re-measurement sometimes differs from the nominal value, which may be due to the position of the lyophilized plasmid in the tube, the efficiency of the re-dissolution, the measurement bias, and adsorption on the wall of the tube, therefore, it is recommended to transform and extract the plasmid before using it
General Plasmid Transform Protocol
1. Take one 100μl of the competent cells and thaw it on ice for 10min, add 2μl of plasmid, then ice bath for 30min, then heat-shock it at 42℃ for 60s, do not stir, and then ice bath for 2min.
2. Add 900μl of LB liquid medium without antibiotics, and incubate at 37℃ for 45min (30℃ for 1-1.5 hours) with 180rpm shaking.
3. Centrifuge at 6000rpm for 5min, leave only 100μl of supernatant to resuspend the bacterial precipitate and spread it onto the target plasmid-resistant LB plate.
4. Invert the plate and incubate at 37℃ for 14h, or at 30℃ for 20h.
5. Pick a single colony into LB liquid medium, add the corresponding antibiotics, incubate at 220rpm for 14h, and extract the plasmid according to the experimental needs and the instructions of the plasmid extraction kit.
Human EEF1A1 ORF clone (NM_001402) (Cat. No.: V035085) Sequence
LOCUS Exported 6826 bp DNA circular SYN 26-FEB-2026
DEFINITION synthetic circular DNA.
ACCESSION V035085
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6826)
AUTHORS .
TITLE Direct Submission
FEATURES Location/Qualifiers
source 1..6826
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 202..581
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 582..785
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
promoter 830..848
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
CDS 931..2316
/codon_start=1
/gene="EEF1A1"
/label=EEF1A1(NM_001402)
/note="EEF1A1(NM_001402)"
/translation="MGKEKTHINIVVIGHVDSGKSTTTGHLIYKCGGIDKRTIEKFEKE
AAEMGKGSFKYAWVLDKLKAERERGITIDISLWKFETSKYYVTIIDAPGHRDFIKNMIT
GTSQADCAVLIVAAGVGEFEAGISKNGQTREHALLAYTLGVKQLIVGVNKMDSTEPPYS
QKRYEEIVKEVSTYIKKIGYNPDTVAFVPISGWNGDNMLEPSANMPWFKGWKVTRKDGN
ASGTTLLEALDCILPPTRPTDKPLRLPLQDVYKIGGIGTVPVGRVETGVLKPGMVVTFA
PVNVTTEVKSVEMHHEALSEALPGDNVGFNVKNVSVKDVRRGNVAGDSKNDPPMEAAGF
TAQVIILNHPGQISAGYAPVLDCHTAHIACKFAELKEKIDRRSGKKLEDGPKFLKSGDA
AIVDMVPGKPMCVESFSDYPPLGRFAVRDMRQTVAVGVIKAVDKKAAGAGKVTKSAQKA
QKAK"
CDS 2323..2352
/codon_start=1
/product="Myc (human c-Myc proto-oncogene) epitope tag"
/label=Myc
/translation="EQKLISEEDL"
polyA_signal 2390..2614
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
rep_origin 2660..3088
/direction=RIGHT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3102..3431
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3498..4292
/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"
polyA_signal 4466..4587
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
primer_bind complement(4636..4652)
/label=M13 rev
/note="common sequencing primer, one of multiple similar
variants"
protein_bind 4660..4676
/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(4684..4714)
/label=lac promoter
/note="promoter for the E. coli lac operon"
protein_bind 4729..4750
/label=CAP binding site
/bound_moiety="E. coli catabolite activator protein"
/note="CAP binding activates transcription in the presence
of cAMP."
rep_origin complement(5038..5626)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5797..6657)
/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"
promoter complement(6658..6762)
/gene="bla"
/label=AmpR promoter