LOCUS       sequence-233935-        5471 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of mCherry and moxMaple3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5471)
  AUTHORS   Kaberniuk AA, Mohr MA, Verkhusha VV, Snapp EL
  TITLE     moxMaple3: a Photoswitchable Fluorescent Protein for PALM and 
            Protein Highlighting in Oxidizing Cellular Environments.
  JOURNAL   Sci Rep. 2018 Oct 3;8(1):14738. doi: 10.1038/s41598-018-32955-5.
  PUBMED    30283009
REFERENCE   2  (bases 1 to 5471)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2018-10-3"; volume: "8(1):14738. doi"; pages: " 
            10.1038/s41598-018-32955-5"
COMMENT     Sequence Label: pmCherry-T2A-moxMaple3
FEATURES             Location/Qualifiers
     source          1..5471
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        54..357
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        358..561
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     512..532
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             618..1325
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(763..781)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1031..1050
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             1344..1397
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     polyA_signal    2252..2373
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2289..2308)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2343..2362
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2380..2835)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2517..2538)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2729..2748
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2862..2966
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2968..3325
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2968..3159
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2991..3011)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3176..3311
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3360..4154
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3360..4154
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3414..3433)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4024..4043
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4342..4361)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4386..4433
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4762..5350
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5251..5270
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 taatagtaat caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca
       61 taacttacgg taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca
      121 ataatgacgt atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg
      181 gagtatttac ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg
      241 ccccctattg acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc
      301 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      361 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      421 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      481 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      541 gaggtctata taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggact
      601 cagatctcga gctcaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg
      661 agttcatgcg cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg
      721 agggcgaggg cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca
      781 agggtggccc cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca
      841 aggcctacgt gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg
      901 gcttcaagtg ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg
      961 actcctccct gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc
     1021 cctccgacgg ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga
     1081 tgtaccccga ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg
     1141 gcggccacta cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc
     1201 ccggcgccta caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca
     1261 tcgtggaaca gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt
     1321 acaaggctcg agataggcgc gccgagggca gaggaagtct gctaacatgc ggtgacgtcg
     1381 aggagaatcc tggcccaact agtatggtga gcaagggcga ggagaccatc atgagcgtga
     1441 tcaagcccga catgaagatc aagctgcgca tggagggcaa cgtgaacggc cacgccttcg
     1501 tgatcgaggg cgagggcagc ggcaagccct tcgagggcat ccagaccatc gacctggagg
     1561 tgaaggaggg cgcccccctg cccttcgcct acgacatcct gaccaccgcc ttccactacg
     1621 gcaaccgcgt gttcaccaag tacccccgca agatccccga ctacttcaag cagagcttcc
     1681 ccgagggcta cagctgggag cgcagcatga cctacgagga cggcggcatc gtccaggcca
     1741 ccaacgacat caccatggag gaggacagct tcatcaacaa gatccacttc aagggcacca
     1801 acttcccccc caacggcccc gtgatgcaga agcgcaccgt gggctgggag gtgagcaccg
     1861 agaagatgta cgtgcgcgac ggcgtgctga agggcgacgt gaagatgaag ctgctgctga
     1921 agggcggcag ccactaccgc gccgacttcc gcaccaccta caaggtgaag cagaaggccg
     1981 tgaagctgcc caaggcccac ttcgtggacc accgcatcga gatcctgagc cacgacaagg
     2041 actacaacaa ggtgaagctg tacgagcacg ccgtggcccg cgacagcacc gacagcatgg
     2101 acgagctgta caagtaaccc gggatccacc ggggccgcga ctctagatca taatcagcca
     2161 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2221 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2281 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2341 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     2401 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     2461 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     2521 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     2581 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     2641 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     2701 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     2761 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     2821 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     2881 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     2941 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3001 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3061 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3121 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3181 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3241 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3301 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3361 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     3421 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     3481 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     3541 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     3601 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     3661 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     3721 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     3781 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     3841 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     3901 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     3961 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4021 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4081 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4141 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4201 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4261 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4321 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4381 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     4441 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     4501 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     4561 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     4621 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     4681 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     4741 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4801 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4861 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4921 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4981 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5041 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5101 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5161 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5221 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5281 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5341 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     5401 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     5461 gcattagtta t
//
