Basic Vector Information
- Vector Name:
- PA-mCherry1-C1
- Antibiotic Resistance:
- Kanamycin
- Length:
- 4716 bp
- Type:
- Mammalian Expression
- Replication origin:
- ori
- Selection Marker:
- Neomycin (select with G418)
- Copy Number:
- High Copy
- Promoter:
- CMV
PA-mCherry1-C1 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 ℃.
PA-mCherry1-C1 vector Sequence
LOCUS 40924_2972 4716 bp DNA circular SYN 13-MAY-2021 DEFINITION Localization: C1 Cloning Vector, Excitation: 400 / 504, Emission: 515 / 517. ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 4716) TITLE Michael Davidson Empty Backbones REFERENCE 2 (bases 1 to 4716) TITLE Direct Submission REFERENCE 3 (bases 1 to 4716) AUTHORS . TITLE Direct Submission COMMENT SGRef: number: 1; type: "Journal Article" COMMENT SGRef: number: 2; type: "Journal Article" FEATURES Location/Qualifiers source 1..4716 /mol_type="other DNA" /organism="synthetic DNA construct" enhancer 115..418 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 419..622 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" CDS 661..1368 /codon_start=1 /label=PAmCherry1 /note="photoactivatable monomeric derivative of DsRed fluorescent protein (Subach et al., 2009)" /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA EGRHSTGGMDELYK" misc_feature 1369..1434 /label=MCS /note="multiple cloning site" polyA_signal 1558..1679 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(1686..2141) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" promoter 2168..2272 /label=AmpR promoter promoter 2274..2631 /label=SV40 promoter /note="SV40 enhancer and early promoter" CDS 2666..3457 /codon_start=1 /label=NeoR/KanR /note="aminoglycoside phosphotransferase" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(3648..3667) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3692..3739 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 4068..4656 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication"
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