Basic Vector Information
- Vector Name:
- (194)_P_EF1a-mCitrine-I
- Antibiotic Resistance:
- Ampicillin
- Length:
- 7342 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Frei T, Cella F, Tedeschi F, Gutierrez J
- Promoter:
- EF-1α
(194)_P_EF1a-mCitrine-I 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 ℃.
(194)_P_EF1a-mCitrine-I vector Sequence
LOCUS 62056_26 7342 bp DNA circular SYN 17-NOV-2020
DEFINITION Cloning vector (194)_P_EF1a-mCitrine-I, complete sequence.
ACCESSION MT891329
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 7342)
AUTHORS Frei T, Cella F, Tedeschi F, Gutierrez J, Stan GB, Khammash M,
Siciliano V.
TITLE Characterization and mitigation of gene expression burden in
mammalian cells
JOURNAL Nat Commun 11 (1), 4641 (2020)
PUBMED 32934213
REFERENCE 2 (bases 1 to 7342)
AUTHORS Frei T, Cella F, Tedeschi F, Gutierrez J, Stan G-B., Khammash M,
Siciliano V.
TITLE Direct Submission
JOURNAL Submitted (15-AUG-2020) Synthetic and Systems Biology for
Biomedicine, Italian Institute of Technology-IIT, Largo Barsanti e
Martteucci 53, Naples, Campania 80125, Italy
REFERENCE 3 (bases 1 to 7342)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Commun"; date: "2020"; volume: "11"; issue: "1"; pages: "4641"
COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted
(15-AUG-2020) Synthetic and Systems Biology for Biomedicine, Italian
Institute of Technology-IIT, Largo Barsanti e Martteucci 53, Naples,
Campania 80125, Italy"
COMMENT ##Assembly-Data-START##
Sequencing Technology :: Sanger dideoxy sequencing
##Assembly-Data-END##
FEATURES Location/Qualifiers
source 1..7342
/mol_type="other DNA"
/organism="synthetic DNA construct"
regulatory 1..266
/note="misc_signal; INS"
/regulatory_class="other"
polyA_signal 283..490
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 523..1704
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 1726..2442
/label=mEYFP
/note="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
regulatory complement(2464..2482)
/label=SP6 promoter
/note="SP6 promoter"
/regulatory_class="promoter"
polyA_signal 2643..2764
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 2854..3143
/note="misc_signal; INS"
/regulatory_class="other"
polyA_signal 3156..3380
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 3415..3731
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3762..4706
/label=ORF frame 1
/note="bright monomeric near-infrared fluorescent protein
engineered from a bacterial phytochrome, with an emission
maximum of 670 nm (Shcherbakova et al., 2016)"
promoter complement(4739..4757)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
regulatory 4918..5039
/label=SV40 poly(A) signal
/note="SV40 poly(A) signal"
/regulatory_class="polyA_signal_sequence"
regulatory 5129..5418
/note="misc_signal; INS"
/regulatory_class="other"
rep_origin complement(5497..6085)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(6259..7116)
/label=AmpR
/note="beta-lactamase"
promoter complement(7117..7221)
/label=AmpR promoter
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