Basic Vector Information
- Vector Name:
- pmiRFP670-N1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4960 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
- Cloning Method:
- Restriction Enzyme
- 5' Primer:
- CMV-F
- 3' Primer:
- SV40pA-R
pmiRFP670-N1 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 ℃.
pmiRFP670-N1 vector Sequence
LOCUS 40924_30905 4960 bp DNA circular SYN 13-MAY-2021 DEFINITION Monomeric near-infrared fluorescent protein miRFP670 in mammalian plasmid. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4960) AUTHORS Shcherbakova DM, Baloban M, Emelyanov AV, Brenowitz M, Guo P, Verkhusha VV TITLE Bright monomeric near-infrared fluorescent proteins as tags and biosensors for multiscale imaging. JOURNAL Nat Commun. 2016 Aug 19;7:12405. doi: 10.1038/ncomms12405. PUBMED 27539380 REFERENCE 2 (bases 1 to 4960) TITLE Direct Submission REFERENCE 3 (bases 1 to 4960) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "Nat Commun."; date: "2016-08-19"; pages: " 10.1038/ncomms12405" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..4960 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 106..409 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 410..613 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" misc_feature 636..716 /label=MCS /note="multiple cloning site" CDS 724..1668 /codon_start=1 /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)" /translation="MVAGHASGSPAFGTASHSNCEHEEIHLAGSIQPHGALLVVSEHDH RVIQASANAAEFLNLGSVLGVPLAEIDGDLLIKILPHLDPTAEGMPVAVRCRIGNPSTE YCGLMHRPPEGGLIIELERAGPSIDLSGTLAPALERIRTAGSLRALCDDTVLLFQQCTG YDRVMVYRFDEQGHGLVFSECHVPGLESYFGNRYPSSTVPQMARQLYVRQRVRVLVDVT YQPVPLEPRLSPLTGRDLDMSGCFLRSMSPCHLQFLKDMGVRATLAVSLVVGGKLWGLV VCHHYLPRFIRFELRAICKRLAERIATRITALES" polyA_signal 1793..1914 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(1921..2376) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2403..2507 /label=AmpR promoter promoter 2509..2866 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2901..3692 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3883..3902) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3927..3974 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4303..4891 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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