LOCUS       sequence-421915-        5694 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of Ebolavirus matrix protein VP40 with an 
            mCherry fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5694)
  AUTHORS   Shoemaker CJ, Schornberg KL, Delos SE, Scully C, Pajouhesh H, 
            Olinger GG, Johansen LM, White JM
  TITLE     Multiple cationic amphiphiles induce a Niemann-Pick C phenotype and 
            inhibit Ebola virus entry and infection.
  JOURNAL   PLoS One. 2013;8(2):e56265. doi: 10.1371/journal.pone.0056265. Epub 
            2013 Feb 18.
  PUBMED    23441171
REFERENCE   2  (bases 1 to 5694)
  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: "PLoS One.";
            date: "2013"; volume: "8(2):e56265. doi"; pages: " 
            10.1371/journal.pone.0056265. Epub 2013 Feb 18"
COMMENT     Sequence Label: mCherry-VP40
FEATURES             Location/Qualifiers
     source          1..5694
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        47..404
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        47..238
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(70..90)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      255..390
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             439..1233
                     /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             439..1233
                     /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(493..512)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1103..1122
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1421..1440)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1465..1512
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1841..2429
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2330..2349
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2604..2907
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2908..3111
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3062..3082
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      3150..3159
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3156..3863
                     /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(3301..3319)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3569..3588
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    5025..5146
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5062..5081)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5116..5135
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5153..5608)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5290..5311)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5502..5521
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        join(5635..5694,1..45)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga ggcggaaaga
       61 accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
      121 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
      181 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
      241 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
      301 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct gagctattcc
      361 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga tcaagagaca
      421 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
      481 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
      541 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
      601 ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc cacgacgggc
      661 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
      721 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
      781 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
      841 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
      901 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
      961 aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     1021 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     1081 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     1141 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     1201 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     1261 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     1321 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     1381 tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg aaggagacaa
     1441 taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg cacggtgttg
     1501 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     1561 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     1621 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcct
     1681 caggttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     1741 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     1801 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     1861 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     1921 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     1981 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     2041 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     2101 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     2161 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     2221 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     2281 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     2341 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     2401 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     2461 tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct gattctgtgg
     2521 ataaccgtat taccgccatg cattagttat taatagtaat caattacggg gtcattagtt
     2581 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     2641 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     2701 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     2761 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     2821 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     2881 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     2941 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     3001 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     3061 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tggtttagtg
     3121 aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt gagcaagggc gaggaggata
     3181 acatggccat catcaaggag ttcatgcgct tcaaggtgca catggagggc tccgtgaacg
     3241 gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc acccagaccg
     3301 ccaagctgaa ggtgaccaag ggtggccccc tgcccttcgc ctgggacatc ctgtcccctc
     3361 agttcatgta cggctccaag gcctacgtga agcaccccgc cgacatcccc gactacttga
     3421 agctgtcctt ccccgagggc ttcaagtggg agcgcgtgat gaacttcgag gacggcggcg
     3481 tggtgaccgt gacccaggac tcctccctgc aggacggcga gttcatctac aaggtgaagc
     3541 tgcgcggcac caacttcccc tccgacggcc ccgtaatgca gaagaagacc atgggctggg
     3601 aggcctcctc cgagcggatg taccccgagg acggcgccct gaagggcgag atcaagcaga
     3661 ggctgaagct gaaggacggc ggccactacg acgctgaggt caagaccacc tacaaggcca
     3721 agaagcccgt gcagctgccc ggcgcctaca acgtcaacat caagttggac atcacctccc
     3781 acaacgagga ctacaccatc gtggaacagt acgaacgcgc cgagggccgc cactccaccg
     3841 gcggcatgga cgagctgtac aagtccggac tcagatctcc ggaattcgcc atgaggcggg
     3901 ttatattgcc tactgctcct cctgaatata tggaggccat ataccctgtc aggtcaaatt
     3961 caacaattgc tagaggtggc aacagcaata caggcttcct gacaccggag tcagtcaatg
     4021 gggacactcc atcgaatcca ctcaggccaa ttgccgatga caccatcgac catgccagcc
     4081 acacaccagg cagtgtgtca tcagcattca tccttgaagc tatggtgaat gtcatatcgg
     4141 gccccaaagt gctaatgaag caaattccaa tttggcttcc tctaggtgtc gctgatcaaa
     4201 agacctacag ctttgactca actacggccg ccatcatgct tgcttcatac actatcaccc
     4261 atttcggcaa ggcaaccaat ccacttgtca gagtcaatcg gctgggtcct ggaatcccgg
     4321 atcatcccct caggctcctg cgaattggaa accaggcttt cctccaggag ttcgttcttc
     4381 cgccagtcca actaccccag tatttcacct ttgatttgac agcactcaaa ctgatcaccc
     4441 aaccactgcc tgctgcaaca tggaccgatg acactccaac aggatcaaat ggagcgttgc
     4501 gtccaggaat ttcatttcat ccaaaacttc gccccattct tttacccaac aaaagtggga
     4561 agaaggggaa cagtgccgat ctaacatctc cggagaaaat ccaagcaata atgacttcac
     4621 tccaggactt taagatcgtt ccaattgatc caaccaaaaa tatcatggga atcgaagtgc
     4681 cagaaactct ggtccacaag ctgaccggta agaaggtgac ttctaaaaat ggacaaccaa
     4741 tcatccctgt tcttttgcca aagtacattg ggttggaccc ggtggctcca ggagacctca
     4801 ccatggtaat cacacaggat tgtgacacgt gtcattctcc tgcaagtctt ccagctgtga
     4861 ttgagaagta actcgtcgac ggtaccgcgg gcccgggatc caccggatct agataactga
     4921 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     4981 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5041 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5101 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttaacgc gtaaattgta
     5161 agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac
     5221 caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg
     5281 agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa
     5341 gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt
     5401 tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt
     5461 agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga
     5521 gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc
     5581 gcgcttaatg cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga
     5641 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gaga
//
