Basic Vector Information
pmCherry-neo4 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 ℃.
pmCherry-neo4 vector Sequence
LOCUS 40924_30001 6116 bp DNA circular SYN 18-DEC-2018 DEFINITION Transformation vector pmCherry-neo4 DNA, complete sequence. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6116) AUTHORS Kataoka K, Schoeberl UE, Mochizuki K. TITLE Modules for C-terminal epitope tagging of Tetrahymena genes JOURNAL J. Microbiol. Methods 82 (3), 342-346 (2010) PUBMED 20624430 REFERENCE 2 (bases 1 to 6116) AUTHORS Kataoka K, Mochizuki K. TITLE Direct Submission JOURNAL Submitted (29-JUN-2010) Contact:Kazufumi Mochizuki Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA); Dr. Bohr-Gasse 3, Vienna A-1030, Austria REFERENCE 3 (bases 1 to 6116) TITLE Direct Submission REFERENCE 4 (bases 1 to 6116) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article"; journalName: "J. Microbiol. Methods"; date: "2010"; volume: "82"; issue: "3"; pages: "342-346" COMMENT SGRef: number: 2; type: "Journal Article"; journalName: "Submitted (29-JUN-2010) Contact:Kazufumi Mochizuki Institute of Molecular Biotechnology of the Austrian Academy of Sciences (IMBA); Dr. Bohr-Gasse 3, Vienna A-1030, Austria" COMMENT SGRef: number: 3; type: "Journal Article" FEATURES Location/Qualifiers source 1..6116 /mol_type="other DNA" /organism="synthetic DNA construct" promoter 21..125 /label=AmpR promoter CDS 126..983 /codon_start=1 /label=AmpR /note="beta-lactamase" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" rep_origin 1157..1745 /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" protein_bind 2033..2054 /label=CAP binding site /note="CAP binding activates transcription in the presence of cAMP." promoter 2069..2099 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 2107..2123 /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)." primer_bind 2131..2147 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" promoter 2168..2186 /label=T3 promoter /note="promoter for bacteriophage T3 RNA polymerase" CDS 2247..2954 /codon_start=1 /label=mCherry /note="monomeric derivative of DsRed fluorescent protein (Shaner et al., 2004)" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" misc_feature 2961..3359 /label=BTU1 3' flanking /note="BTU1 3' flanking" CDS 4271..5062 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5431..5447) /label=KS primer /note="common sequencing primer, one of multiple similar variants" promoter complement(5473..5491) /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" primer_bind complement(5498..5514) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" rep_origin 5656..6111 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis"
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