pmCherry-EGFP-LC3b
Catalog No. PVT10398
Packing 2ug
pmCherry-EGFP-LC3b Information
Function ORF expression plasmid
Promoter: CMV
Replicator: pUC ori, F1 ori, SV40 ori
Terminator: SV40 poly (A) signal
Plasmid classification: mammalian cells, autophagic expression vectors
Plasmid size: 5833bp
Prokaryotic resistance: kanamycin Kan
Screening markers: Mycophenolate G418
Cloned strain: Escherichia coli 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: Sv40-polyA-R:GAAATTTGTGATGCTATTGC
pmCherry-EGFP-LC3b Description
PmCherry-EGFP-LC3b is a double fluorescent reporter plasmid for human mammalian cell autophagy.
pmCherry-EGFP-LC3b Reference
Foot-and-mouth disease virus capsid protein VP2 activates the cellular EIF2S1-ATF4 pathway and induces autophagy via HSPB1.
pmCherry-EGFP-LC3b Sequence
LOCUS Exported 5833 bp ds-DNA circular SYN 12-MAY-2016
FEATURES Location/Qualifiers
source 1..5833
/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"
CDS 604..1311
/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"
CDS 1333..2049
/codon_start=1
/product="enhanced GFP"
/note="EGFP"
/note="mammalian codon-optimized"
/translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
EFVTAAGITLGMDELYK"
misc_feature 2083..2457
/note="LC3B-h"
polyA_signal 2621..2742
/note="SV40 poly(A) signal"
/note="SV40 polyadenylation signal"
rep_origin complement(2749..3204)
/direction=LEFT
/note="f1 ori"
/note="f1 bacteriophage origin of replication; arrow
indicates direction of (+) strand synthesis"
promoter 3231..3335
/gene="bla"
/note="AmpR promoter"
promoter 3337..3694
/note="SV40 promoter"
/note="SV40 enhancer and early promoter"
rep_origin 3545..3680
/note="SV40 ori"
/note="SV40 origin of replication"
CDS 3729..4523
/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 4755..4802
/note="HSV TK poly(A) signal"
/note="herpes simplex virus thymidine kinase
polyadenylation signal (Cole and Stacy, 1985)"
rep_origin 5131..5719
/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 gctagccgcc
601 accatggtga gcaagggcga ggaggataac atggccatca tcaaggagtt catgcgcttc
661 aaggtgcaca tggagggctc cgtgaacggc cacgagttcg agatcgaggg cgagggcgag
721 ggccgcccct acgagggcac ccagaccgcc aagctgaagg tgaccaaggg tggccccctg
781 cccttcgcct gggacatcct gtcccctcag ttcatgtacg gctccaaggc ctacgtgaag
841 caccccgccg acatccccga ctacttgaag ctgtccttcc ccgagggctt caagtgggag
901 cgcgtgatga acttcgagga cggcggcgtg gtgaccgtga cccaggactc ctccctgcag
961 gacggcgagt tcatctacaa ggtgaagctg cgcggcacca acttcccctc cgacggcccc
1021 gtaatgcaga agaagaccat gggctgggag gcctcctccg agcggatgta ccccgaggac
1081 ggcgccctga agggcgagat caagcagagg ctgaagctga aggacggcgg ccactacgac
1141 gctgaggtca agaccaccta caaggccaag aagcccgtgc agctgcccgg cgcctacaac
1201 gtcaacatca agttggacat cacctcccac aacgaggact acaccatcgt ggaacagtac
1261 gaacgcgccg agggccgcca ctccaccggc ggcatggacg agctgtacaa gtccggactc
1321 agatctcgag gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
1381 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
1441 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
1501 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
1561 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
1621 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
1681 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
文献和实验
技术资料