LOCUS       sequence-447476-        7795 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Ebola virus minigenome with firefly luciferase reporter 
            gene. Minigenome is under control of the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7795)
  AUTHORS   Nelson EV, Pacheco JR, Hume AJ, Cressey TN, Deflube LR, Ruedas JB, 
            Connor JH, Ebihara H, Muhlberger E
  TITLE     An RNA polymerase II-driven Ebola virus minigenome system as an 
            advanced tool for antiviral drug screening.
  JOURNAL   Antiviral Res. 2017 Aug 12;146:21-27. doi: 
            10.1016/j.antiviral.2017.08.005.
  PUBMED    28807685
REFERENCE   2  (bases 1 to 7795)
  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: "Antiviral 
            Res."; date: "2017-08-12"; volume: "146:21-27. doi"; pages: " 
            10.1016/j.antiviral.2017.08.005"
COMMENT     Sequence Label: pCAGGS_3E5E_luciferase
FEATURES             Location/Qualifiers
     source          1..7795
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..666
                     /label=chicken beta-actin promoter
     intron          667..1683
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1607..1629
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1691..1710
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             complement(2574..4226)
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     4142..4160
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(4817..4836)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4882..4937
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4936..4955)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5298..5314)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5298..5314)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5311..5333)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5322..5338
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5346..5352,5353..5370,5371..5376))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5391..5412
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5471..5666
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5517..5652
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5672..5806
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5709..5728)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5763..5782
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5874..5891)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6045..6633)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6125..6144)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6804..7595,7596..7664))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7427..7446
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7665..7769)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc ccctccccac ccccaatttt gtatttattt attttttaat tattttgtgc
      481 agcgatgggg gcgggggggg gggggggggg cgcgcgccag gcggggcggg gcggggcgag
      541 gggcggggcg gggcgaggcg gagaggtgcg gcggcagcca atcagagcgg cgcgctccga
      601 aagtttcctt ttatggcgag gcggcggcgg cggcggccct ataaaaagcg aagcgcgcgg
      661 cgggcgggag tcgctgcgcg ctgccttcgc cccgtgcccc gctccgccgc cgcctcgcgc
      721 cgcccgcccc ggctctgact gaccgcgtta ctcccacagg tgagcgggcg ggacggccct
      781 tctcctccgg gctgtaatta gcgcttggtt taatgacggc ttgtttcttt tctgtggctg
      841 cgtgaaagcc ttgaggggct ccgggagggc cctttgtgcg gggggagcgg ctcggggggt
      901 gcgtgcgtgt gtgtgtgcgt ggggagcgcc gcgtgcggct ccgcgctgcc cggcggctgt
      961 gagcgctgcg ggcgcggcgc ggggctttgt gcgctccgca gtgtgcgcga ggggagcgcg
     1021 gccgggggcg gtgccccgcg gtgcgggggg ggctgcgagg ggaacaaagg ctgcgtgcgg
     1081 ggtgtgtgcg tgggggggtg agcagggggt gtgggcgcgt cggtcgggct gcaacccccc
     1141 ctgcaccccc ctccccgagt tgctgagcac ggcccggctt cgggtgcggg gctccgtacg
     1201 gggcgtggcg cggggctcgc cgtgccgggc ggggggtggc ggcaggtggg ggtgccgggc
     1261 ggggcggggc cgcctcgggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
     1321 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
     1381 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
     1441 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
     1501 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
     1561 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
     1621 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
     1681 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcctc
     1741 gagggtaccc ccggggcggc cgcgagctct taattagcta gcgaattgtg tgtccactga
     1801 tgagtccgtg aggacgaaac ccggagtccc gggtctggac acacaaaaaa gaagaaatag
     1861 atttattttt aaatttttgt gtgcgaccat ttttccagga atcctttttg caacgtttat
     1921 tctggcgagc cttctgatga gcgtggtcaa tgtcttaatt atcattaaca cgaagattat
     1981 tatctagtct gctatattgt aaagaaggta tttctgattt tacagtcctg ccttttcttt
     2041 taattttatc attctgtatg aaatagtagc ttactagaca aaaatgttaa ggttattaca
     2101 cctaatctct tattttacac aaagattaag gctatcaccg tttttaaaac gacaaattaa
     2161 agtaatgata attataaaaa acttaatctt aaagatcaat tttgttttaa gaagttgtgt
     2221 tatcaaccaa aacactattc catctgacag gatattgata caacattatt atatcaataa
     2281 caatatgagc ccagaccttt cgttaaagct taattataaa gagtgcagag tttattatgt
     2341 tgcgttaaat tcattgcggc agtaatatgt cacacatgct gcattgtgaa ttttttgatt
     2401 atcacgcagg gagagaggct aaatatagta tataatatag ggactccttt cgttatttaa
     2461 atagttgttt attgagatta gatcaatatg atgtatgaga gcaatttatg agttttttat
     2521 aatctgtatg ttaataacca acctttgcaa gtatcaggat actatgcggc cgcttacacg
     2581 gcgatctttc cgcccttctt ggcctttatg aggatctctc tgatttttct tgcgtcgagt
     2641 tttccggtaa gacctttcgg tacttcgtcc acaaacacaa ctcctccgcg caactttttc
     2701 gcggttgtta cttgactggc gacgtaatcc acgatctctt tttccgtcat cgtctttccg
     2761 tgctccaaaa caacaacggc ggcgggaagt tcaccggcgt catcgtcggg aagacctgcg
     2821 acacctgcgt cgaagatgtt ggggtgttgg agcaagatgg attccaattc agcgggagcc
     2881 acctgatagc ctttgtactt aatcagagac ttcaggcggt caacgatgaa gaagtgttcg
     2941 tcttcgtccc agtaagctat gtctccagaa tgtagccatc catccttgtc aatcaaggcg
     3001 ttggtcgctt ccggattgtt tacataaccg gacataatca taggacctct cacacacagt
     3061 tcgcctcttt gattaacgcc cagcgttttc ccggtatcca gatccacaac cttcgcttca
     3121 aaaaatggaa caactttacc gaccgcgccc ggtttatcat ccccctcggg tgtaatcaga
     3181 atagctgatg tagtctcagt gagcccatat ccttgcctga tacctggcag atggaacctc
     3241 ttggcaaccg cttccccgac ttccttagag aggggagcgc caccagaagc aatttcgtgt
     3301 aaattagata aatcgtattt gtcaatcaga gtgcttttgg cgaagaagga gaatagggtt
     3361 ggcaccagca gcgcactttg aatcttgtaa tcctgaaggc tcctcagaaa cagctcttct
     3421 tcaaatctat acattaagac gactcgaaat ccacatatca aatatccgag tgtagtaaac
     3481 attccaaaac cgtgatggaa tggaacaaca cttaaaatcg cagtatccgg aatgatttga
     3541 ttgccaaaaa taggatctct ggcatgcgag aatctcacgc aggcagttct atgaggcaga
     3601 gcgacacctt taggcagacc agtagatcca gaggagttca tgatcagtgc aattgtcttg
     3661 tccctatcga aggactctgg cacaaaatcg tattcattaa aaccgggagg tagatgagat
     3721 gtgacgaacg tgtacatcga ctgaaatccc tggtaatccg ttttagaatc catgataata
     3781 attttttgga tgattgggag ctttttttgc acgttcaaaa ttttttgcaa cccctttttg
     3841 gaaacgaaca ccacggtagg ctgcgaaatg cccatactgt tgagcaattc acgttcatta
     3901 taaatgtcgt tcgcgggcgc aactgcaact ccgataaata acgcgcccaa caccggcata
     3961 aagaattgaa gagagttttc actgcatacg acgattctgt gatttgtatt cagcccatat
     4021 cgtttcatag cttctgccaa ccgaacggac atttcgaagt actcagcgta agtgatgtcc
     4081 acctcgatat gtgcatctgt aaaagcaatt gttccaggaa ccagggcgta tctcttcata
     4141 gccttatgca gttgctctcc agcggttcca tcttccagcg gatagaatgg cgccgggcct
     4201 ttctttatgt ttttggcgtc ttccatatga gacatactcg gaattttgtg attccgagca
     4261 atttgtcttt taactatttc caatgtttta ccccaagctt tagggttgtt gaatcttcaa
     4321 tcccctaagt gttcaagttc aaaaccaagg ttaattctca atcttgcttg tttgactgtg
     4381 aactaatgct gtcctttaac aataatgttt ctattaaggt tgttggatca ccctgcagag
     4441 tctgaggtgg aacttgtgat gtgataagac ctaggtgttg acaagggatt ttcaactgag
     4501 cacactgatg cccttgctcc cttcggagac ctaggttgtt ctctatcttt gtgatgtggc
     4561 tctgaaacaa accaggtgtg attacagtaa caatttcaat ttaaattccg atataaattt
     4621 caatgagagg aaaattatta atcttcctca tagttattcg cacacaaaag atcctaaaaa
     4681 ttcttctttc tttttgtgtg tccggggtcg gcatggcatc tccacctcct cgcggtccga
     4741 cctgggcatc cgaaggagga cgtcgtccac tcggatggct aagggagagc tccctagatc
     4801 ggaactcgct agcagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc
     4861 ttgagcatct gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga
     4921 attttttgtg tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga
     4981 atgagtattt ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag
     5041 gttggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     5101 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     5161 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     5221 gactactccc agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca
     5281 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     5341 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     5401 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     5461 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     5521 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5581 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5641 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     5701 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5761 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     5821 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5881 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5941 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     6001 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     6061 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     6121 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     6181 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     6241 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     6301 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     6361 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     6421 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     6481 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     6541 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     6601 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6661 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6721 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6781 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6841 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6901 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6961 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     7021 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     7081 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     7141 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     7201 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     7261 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     7321 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     7381 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     7441 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     7501 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     7561 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     7621 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7681 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7741 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
