pmCherry-N1 Search name
pmCherry-N1,Plasmid pmCherry-N1,pmCherry-N1 vector
pmCherry-N1 Information
Promoter: CMV
Replicator: pUC ori, F1 ori
Terminator: SV40 poly (A) signal
Plasmid classification: mammalian cells, fluorescent protein reporting carriers
Plasmid size: 4722bp
Prokaryotic resistance: Kan
Screening markers: Neo
Cloned strain: DH5 alpha
Culture conditions: 37 centigrade, aerobic LB
Expression host: mammalian cells
Induction mode: no induction, instantaneous expression
5'sequencing primers: CMV-F:CGCAAATGGGCGGTAGGCGTG
3'sequencing primers: mCherry-R:TTGGTCACCTTCAGCTTGG
pmCherry-N1 Desciption
pmCherry-N1 is a mammalian expression vector designed to express a protein of interestfused to the N-terminus of mCherry, a mutant fluorescent protein derived from thetetrameric Discosoma sp. red fluorescent protein, DsRed (1). The excitation and emission maxima of the native mCherry protein are 587 nm and 610 nm, respectively. Expression of fusion proteins that retain the fluorescent properties of the unmodified mCherry protein can be monitored by flow cytometry and their localization in vivo can be determined by fluorescence microscopy.
pmCherry-N1 Sequence
LOCUS Exported 4722 bp ds-DNA circular SYN 22-10-2015
DEFINITION .
ACCESSION .
VERSION .
KEYWORDS Untitled 4
SOURCE synthetic DNA construct
ORGANISM synthetic DNA construct
REFERENCE 1 (bases 1 to 4722)
AUTHORS admin
TITLE Direct Submission
JOURNAL Exported 2015-10-22
FEATURES Location/Qualifiers
source 1..4722
/organism="synthetic DNA construct"
/mol_type="other DNA"
enhancer 61..364
/note="CMV enhancer"
/note="human cytomegalovirus immediate early enhancer"
promoter 365..568
/note="CMV promoter"
/note="human cytomegalovirus (CMV) immediate early
promoter"
misc_feature 591..671
/note="MCS"
/note="multiple cloning site"
CDS 679..1389
/codon_start=1
/product="monomeric derivative of DsRed fluorescent protein
(Shaner et al., 2004)"
/note="mCherry"
/note="mammalian codon-optimized"
/translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
EGRHSTGGMDELYK"
polyA_signal 1510..1631
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(1638..2093)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 2120..2224
/gene="bla"
/note="AmpR promoter"
promoter 2226..2583
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 2434..2569
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 2618..3412
/codon_start=1
/gene="aph(3')-II (or nptII)"
/product="aminoglycoside phosphotransferase from Tn5"
/note="NeoR/KanR"
/note="confers resistance to neomycin, kanamycin, and G418
(Geneticin(R))"
/translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
polyA_signal 3644..3691
/note="HSV TK poly(A) signal"
/note="herpesvirus thymidine kinase polyadenylation signal"
rep_origin 4020..4608
/direction=RIGHT
/note="ori"
/note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
replication"
ORIGIN
1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
601 ccggactcag atctcgagct caagcttcga attctgcagt cgacggtacc gcgggcccgg
661 gatccaccgg tcgccaccat ggtgagcaag ggcgaggagg ataacatggc catcatcaag
721 gagttcatgc gcttcaaggt gcacatggag ggctccgtga acggccacga gttcgagatc
781 gagggcgagg gcgagggccg cccctacgag ggcacccaga ccgccaagct gaaggtgacc
841 aagggtggcc ccctgccctt cgcctgggac atcctgtccc ctcagttcat gtacggctcc
901 aaggcctacg tgaagcaccc cgccgacatc cccgactact tgaagctgtc cttccccgag
961 ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg gcgtggtgac cgtgacccag
1021 gactcctccc tgcaggacgg cgagttcatc tacaaggtga agctgcgcgg caccaacttc
1081 ccctccgacg gccccgtaat gcagaagaag accatgggct gggaggcctc ctccgagcgg
1141 atgtaccccg aggacggcgc cctgaagggc gagatcaagc agaggctgaa gctgaaggac
1201 ggcggccact acgacgctga ggtcaagacc acctacaagg ccaagaagcc cgtgcagctg
1261 cccggcgcct acaacgtcaa catcaagttg gacatcacct cccacaacga ggactacacc
1321 atcgtggaac agtacgaacg cgccgagggc cgccactcca ccggcggcat ggacgagctg
1381 tacaagtagc ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta
1441 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
1501 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
1561 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
1621 aatgtatctt aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
1681 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
1741 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
1801 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
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