Basic Vector Information
- Vector Name:
- Clover-mRuby2-FRET-10
- Antibiotic Resistance:
- Kanamycin
- Length:
- 5395 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- SV40
Clover-mRuby2-FRET-10 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 ℃.
Clover-mRuby2-FRET-10 vector Sequence
LOCUS 40924_495 5395 bp DNA circular SYN 13-MAY-2021
DEFINITION Localization: Cytosolic FRET, Excitation: , Emission: .
ACCESSION .
VERSION .
KEYWORDS .
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 5395)
TITLE FRET Indicator - Live and Fixed Cells
REFERENCE 2 (bases 1 to 5395)
TITLE Direct Submission
REFERENCE 3 (bases 1 to 5395)
AUTHORS .
TITLE Direct Submission
COMMENT SGRef: number: 1; type: "Journal Article"
COMMENT SGRef: number: 2; type: "Journal Article"
FEATURES Location/Qualifiers
source 1..5395
/mol_type="other DNA"
/organism="synthetic DNA construct"
enhancer 1..304
/label=CMV enhancer
/note="human cytomegalovirus immediate early enhancer"
promoter 305..508
/label=CMV promoter
/note="human cytomegalovirus (CMV) immediate early
promoter"
CDS 541..1251
/codon_start=1
/label=mRuby2
/note="monomeric red fluorescent protein derived from
mRuby, with improved photophysical properties (Lam et al.,
2012)"
/translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
AKFAGLGGGMDELYK"
CDS 1282..1998
/codon_start=1
/label=Clover
/note="bright green-yellow fluorescent protein derived from
GFP (Lam et al., 2012)"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
KFICTTGKLPVPWPTLVTTFGYGVACFSRYPDHMKQHDFFKSAMPEGYVQERTISFKDD
GTYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIK
ANFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSHQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
polyA_signal 2123..2244
/label=SV40 poly(A) signal
/note="SV40 polyadenylation signal"
rep_origin complement(2251..2706)
/direction=LEFT
/label=f1 ori
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2733..2837
/label=AmpR promoter
promoter 2839..3196
/label=SV40 promoter
/note="SV40 enhancer and early promoter"
CDS 3231..4022
/codon_start=1
/label=NeoR/KanR
/note="aminoglycoside phosphotransferase"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
primer_bind complement(4213..4232)
/label=TK-pA-R
/note="Thymidine kinase polyA, reverse primer"
polyA_signal 4257..4304
/label=HSV TK poly(A) signal
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 4633..5221
/label=ori
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
promoter 5336..5395
/label=CMV
/note="Human cytomegalovirus immediate early
enhancer/promoter"
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