Basic Vector Information
- Vector Name:
- (218)_P_TRE-(HDV)mCitri
- Antibiotic Resistance:
- Kanamycin
- Length:
- 9978 bp
- Type:
- Cloning vector
- Replication origin:
- ori
- Source/Author:
- Frei T, Cella F, Tedeschi F, Gutierrez J
- Promoter:
- EF-1α
(218)_P_TRE-(HDV)mCitri 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 ℃.
(218)_P_TRE-(HDV)mCitri vector Sequence
LOCUS 62056_31 9978 bp DNA circular SYN 17-NOV-2020
DEFINITION Cloning vector (218)_P_TRE-(HDV)mCitri, complete sequence.
ACCESSION MT891330
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 9978)
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 9978)
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 9978)
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..9978
/mol_type="other DNA"
/organism="synthetic DNA construct"
repeat_region 1..35
/label=piggyBac left (5') inverted repeat
/note="piggyBac left (5') inverted repeat"
misc_feature 361..385
/label=ConS scar
/note="ConS scar"
misc_feature 390..414
/label=bidirectional promoter artefact
/note="bidirectional promoter artefact"
promoter 415..729
/label=tight TRE promoter
/note="Tet-responsive promoter PTight, consisting of seven
tet operator sequences followed by the minimal CMV
promoter"
misc_feature 767..919
/label=HDV
/note="HDV"
CDS 917..1630
/label=mEYFP
/note="enhanced YFP with monomerizing A206K mutation
(Zacharias et al., 2002)"
regulatory 1668..1789
/label=SV40 poly(A) signal
/note="SV40 poly(A) signal"
/regulatory_class="polyA_signal_sequence"
misc_feature 1891..1915
/label=Con1 scar
/note="Con1 scar"
regulatory 1954..2219
/note="misc_signal; INS"
/regulatory_class="other"
polyA_signal 2236..2460
/label=bGH poly(A) signal
/note="bovine growth hormone polyadenylation signal"
promoter 2461..3642
/label=EF-1-alpha promoter
/note="strong constitutive promoter for human elongation
factor EF-1-alpha"
CDS 3665..4408
/label=tTA-Advanced
/note="improved tetracycline-controlled transactivator"
CDS 4484..5194
/label=mRuby3
/note="monomeric red fluorescent protein derived from
mRuby2, with improved brightness and photostability (Bajar
et al., 2016)"
polyA_signal complement(5232..5353)
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
misc_feature 5455..5479
/label=ConE scar
/note="ConE scar"
promoter 5499..5828
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
promoter 5876..5923
/label=EM7 promoter
/note="synthetic bacterial promoter"
CDS 5942..6538
/label=PuroR
/note="puromycin N-acetyltransferase"
CDS 6545..7537
/label=Delta-TK
/note="truncated herpes simplex virus thymidine kinase
(Salomon et al., 1995)"
promoter complement(7567..7585)
/label=SP6 promoter
/note="promoter for bacteriophage SP6 RNA polymerase"
polyA_signal 7746..7867
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
repeat_region 8064..8126
/label=piggyBac right (3') inverted repeat
/note="piggyBac right (3') inverted repeat"
regulatory complement(8175..8201)
/label=KanR terminator
/note="KanR terminator"
/regulatory_class="terminator"
CDS complement(8205..9017)
/label=KanR
/note="aminoglycoside phosphotransferase"
regulatory complement(9018..9165)
/label=KanR promoter
/note="KanR promoter"
/regulatory_class="promoter"
rep_origin 9242..9830
/direction=RIGHT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
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