Basic Vector Information
- Vector Name:
- (220)_P_EF1a-mCitrine-T
- Antibiotic Resistance:
- Ampicillin
- Length:
- 8543 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Frei T, Cella F, Tedeschi F, Gutierrez J
- Promoter:
- EF-1α
(220)_P_EF1a-mCitrine-T 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 ℃.
(220)_P_EF1a-mCitrine-T vector Sequence
LOCUS 62056_41 8543 bp DNA circular SYN 17-NOV-2020
DEFINITION Cloning vector (220)_P_EF1a-mCitrine-T, complete sequence.
ACCESSION MT891332
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 8543)
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 8543)
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 8543)
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..8543
/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 526..1707
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 1729..2445
/label=mEYFP
/note="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
misc_binding 2459..2480
/label=miR-FF5 binding site
/bound_moiety="miR-FF5"
/note="mir FF5 target"
misc_binding 2481..2502
/label=miR-FF5 binding site
/bound_moiety="miR-FF5"
/note="mir FF5 target"
misc_binding 2503..2524
/label=miR-FF5 binding site
/bound_moiety="miR-FF5"
/note="mir FF5 target"
regulatory complement(2561..2579)
/label=SP6 promoter
/note="SP6 promoter"
/regulatory_class="promoter"
polyA_signal 2740..2861
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
regulatory 3000..3265
/note="misc_signal; INS"
/regulatory_class="other"
polyA_signal 3282..3506
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 3527..4709
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
misc_feature 4718..4727
/label=Kozak sequence
/note="Kozak sequence"
ncRNA 5491..5619
/label=3' miR-30a
/note="sequence downstream of the 71-nt precursor of the
human miR-30a microRNA (Zeng et al., 2002)"
misc_binding 5828..5849
/label=miR-FF5 binding site
/bound_moiety="miR-FF5"
/note="mir FF5 target"
misc_binding 5850..5871
/label=miR-FF5 binding site
/bound_moiety="miR-FF5"
/note="mir FF5 target"
misc_binding 5872..5893
/label=miR-FF5 binding site
/bound_moiety="miR-FF5"
/note="mir FF5 target"
promoter complement(5931..5949)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
regulatory 6110..6231
/label=SV40 poly(A) signal
/note="SV40 poly(A) signal"
/regulatory_class="polyA_signal_sequence"
regulatory 6321..6610
/note="misc_signal; INS"
/regulatory_class="other"
rep_origin complement(6698..7286)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(7460..8317)
/label=AmpR
/note="beta-lactamase"
promoter complement(8318..8422)
/label=AmpR promoter
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