Basic Vector Information
- Vector Name:
- pTriEx-mCherry-LOV2
- Antibiotic Resistance:
- Ampicillin
- Length:
- 6088 bp
- Type:
- Mammalian Expression, Bacterial Expression
- Replication origin:
- ori
- Copy Number:
- High Copy
- Promoter:
- CMV
- 5' Primer:
- aagtatcgggccctttgtgc
- 3' Primer:
- GGCAGCCTGCACCTGAGGTTAATCAC
pTriEx-mCherry-LOV2 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 ℃.
pTriEx-mCherry-LOV2 vector Sequence
LOCUS 40924_44237 6088 bp DNA circular SYN 13-MAY-2021
DEFINITION mammalian expression for wt LOV2.
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 6088)
AUTHORS Wang H, Vilela M, Winkler A, Tarnawski M, Schlichting I, Yumerefendi
H, Kuhlman B, Liu R, Danuser G, Hahn KM
TITLE LOVTRAP: an optogenetic system for photoinduced protein
dissociation.
JOURNAL Nat Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926.
PUBMED 27427858
REFERENCE 2 (bases 1 to 6088)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 6088)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat
Methods. 2016 Jul 18. doi: 10.1038/nmeth.3926."
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..6088
/mol_type="other DNA"
/organism="synthetic DNA construct"
misc_recomb 110..937
/label=baculovirus recombination region (lef2/ORF603)
/note="contains ORF603 and part of lef2"
enhancer 1005..1308
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 1310..1509
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
primer_bind 1715..1737
/label=pCAGGS-5
/note="Chimeric intron in CAG promoter, forward primer"
primer_bind 1799..1818
/label=pCAG-F
/note="Rabbit beta-globin intron, for pCAG plasmids,
forward primer"
promoter 1860..1878
/label=T7 promoter
/note="promoter for bacteriophage T7 RNA polymerase"
protein_bind 1879..1903
/label=lac operator
/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 1919..2028
/label=p10 promoter
/note="baculovirus promoter for expression in insect cells"
CDS 2043..2750
/codon_start=1
/label=mCherry
/note="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
CDS 3231..3263
/codon_start=1
/label=HSV tag
/note="HSV (herpes simplex virus) epitope tag"
/translation="QPELAPEDPED"
CDS 3270..3293
/codon_start=1
/label=8xHis
/note="8xHis affinity tag"
/translation="HHHHHHHH"
primer_bind complement(3383..3402)
/label=Bglob-pA-R
/note="Rabbit beta-globin polyA region, reverse primer"
polyA_signal 3448..3503
/label=beta-globin poly(A) signal
/note="rabbit beta-globin polyadenylation signal (Gil and
Proudfoot, 1987)"
primer_bind complement(3502..3521)
/label=rbglobpA-R
/note="Rabbit beta-globin polyA, reverse primer. Also
called rb-glob-pA-term-R"
terminator 3591..3638
/label=T7 terminator
/note="transcription terminator for bacteriophage T7 RNA
polymerase"
misc_recomb 3651..4356
/label=baculovirus recombination region (ORF1629)
/note="contains part of ORF1629"
protein_bind complement(4365..4398)
/label=loxP
/note="Cre-mediated recombination occurs in the 8-bp core
sequence (ATGTATGC) (Shaw et al., 2021)."
rep_origin complement(4566..5153)
/direction=LEFT
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
CDS complement(5169..6026)
/codon_start=1
/label=AmpR
/note="beta-lactamase"
/translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
LIKHW"
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