pmCherry- EGFP- LC3B- h/ pmCherry- EGFP- LC3B- h质粒/ pmCherry- EGFP- LC3B- h Plasmid

pmCherry-EGFP-LC3B-h

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Search name

pmCherry-EGFP-LC3B-h,Plasmid pmCherry-EGFP-LC3B-h,pmCherry-EGFP-LC3B-h vector

 

pmCherry-EGFP-LC3B-h Information

 

Alias: mRFP-GFP-LC3; MAP1LC3B; LC3B; ATG8F;

Promoter: CMV

Replicon: pUC ori, F1 ori, SV40 ori

Terminator: SV40 poly (A) signal

Plasmid classification: mammalian cells, autophagic expression vectors

Plasmid size: 5833 BP

Prokaryotic resistance: kanamycin

Screening Marker: Hereditary Mycin G418

Cloning strain: DH5alpha Escherichia coli

Culture conditions: 37鈩? aerobic, LB

Expression host: mammalian cells

Induction mode: no induction, transient expression

5'Sequencing Primers: CMV-F: CGCAAATGGGGGGGTAGGGGGGTG

3'Sequencing primers: Sv40-polyA-R: GAAATTTGATGCTATTGC

 

pmCherry-EGFP-LC3B-h Description

PmCherry-EGFP-LC3b is a human mammalian autophagy double fluorescent reporter plasmid.

 

pmCherry-EGFP-LC3B-h 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

     1741 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac

     1801 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc

     1861 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg

     1921 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc

     1981 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag

     2041 ctgtacaagt actcagatct cgagctcaag cttcgaattc ccatgccgtc ggagaagacc

     2101 ttcaagcagc gccgcacctt cgaacaaaga gtagaagatg tccgacttat tcgagagcag

     2161 catccaacca aaatcccggt gataatagaa cgatacaagg gtgagaagca gcttcctgtt

     2221 ctggataaaa caaagttcct tgtacctgac catgtcaaca tgagtgagct catcaagata

     2281 attagaaggc gcttacagct caatgctaat caggccttct tcctgttggt gaacggacac

     2341 agcatggtca gcgtctccac accaatctca gaggtgtatg agagtgagaa agatgaagat

     2401 ggattcctgt acatggtcta tgcctcccag gagacgttcg ggatgaaatt gtcagtgtaa

     2461 gaattctgca gtcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat

     2521 aatcagccat accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc

     2581 cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta

     2641 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact

     2701 gcattctagt tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg

     2761 ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat

     2821 aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg

     2881 ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc

     2941 gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt

     3001 tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag

     3061 cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg

     3121 gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc

     3181 ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc

     3241 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct

     3301 gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg

     3361 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca

     3421 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg

     3481 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc

     3541 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat

     3601 tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg

     3661 aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat

     3721 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga

     3781 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc

     3841 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga

     3901 atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg

     3961 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc

     4021 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg

     4081 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga

     4141 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc

     4201 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca

     4261 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg

     4321 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct

     4381 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg

     4441 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc

     4501 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac

     4561 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt

     4621 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga

     4681 gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg

     4741 aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt

     4801 tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca

     4861 ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga

     4921 aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc

     4981 atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat

     5041 cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc

     5101 agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg

     5161 ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct

     5221 accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct

     5281 tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct

     5341 cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg

     5401 gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc

     5461 gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga

     5521 gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg

     5581 cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta

     5641 tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg

     5701 ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg

     5761 ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat

     5821 taccgccatg cat

//

 

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