LOCUS       sequence-200889-        6089 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Encodes Ebola virus minigenome with EGFP reporter gene. Minigenome 
            is under control of the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6089)
  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 6089)
  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_eGFP
FEATURES             Location/Qualifiers
     source          1..6089
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     827..849
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     911..930
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             complement(join(1794..2507,2508..2510,2511..2513))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(1797..2507,2508..2510,2511..2513))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1839..1860)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2188..2207
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2447..2468
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3107..3126)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3172..3227
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3226..3245)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3588..3604)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3588..3604)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3601..3623)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3612..3628
                     /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(3636..3642,3643..3660,3661..3666))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3681..3702
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3761..3956
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3807..3942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3962..4096
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3999..4018)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4053..4072
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4164..4181)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4335..4923)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4415..4434)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5094..5885,5886..5954))
                     /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     5717..5736
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5955..6059)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtcgaggtga gccccacgtt ctgcttcact ctccccatct
      421 cccccccctc cccaccccca attttgtatt tatttatttt ttaattattt tgtgcagcga
      481 tgggggcggg gggggggggc cggggagggc tcgggggagg ggcgcggcgg cccccggagc
      541 gccggcggct gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag
      601 agggcgcagg gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc
      661 cgcaccccct ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc
      721 ggggagggcc ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc
      781 tgtccgcggg gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg
      841 cgtgtgaccg gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta
      901 cagctcctgg gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattcctc
      961 gagggtaccc ccggggcggc cgcgagctct taattagcta gcgaattgtg tgtccactga
     1021 tgagtccgtg aggacgaaac ccggagtccc gggtctggac acacaaaaaa gaagaaatag
     1081 atttattttt aaatttttgt gtgcgaccat ttttccagga atcctttttg caacgtttat
     1141 tctggcgagc cttctgatga gcgtggtcaa tgtcttaatt atcattaaca cgaagattat
     1201 tatctagtct gctatattgt aaagaaggta tttctgattt tacagtcctg ccttttcttt
     1261 taattttatc attctgtatg aaatagtagc ttactagaca aaaatgttaa ggttattaca
     1321 cctaatctct tattttacac aaagattaag gctatcaccg tttttaaaac gacaaattaa
     1381 agtaatgata attataaaaa acttaatctt aaagatcaat tttgttttaa gaagttgtgt
     1441 tatcaaccaa aacactattc catctgacag gatattgata caacattatt atatcaataa
     1501 caatatgagc ccagaccttt cgttaaagct taattataaa gagtgcagag tttattatgt
     1561 tgcgttaaat tcattgcggc agtaatatgt cacacatgct gcattgtgaa ttttttgatt
     1621 atcacgcagg gagagaggct aaatatagta tataatatag ggactccttt cgttatttaa
     1681 atagttgttt attgagatta gatcaatatg atgtatgaga gcaatttatg agttttttat
     1741 aatctgtatg ttaataacca acctttgcaa gtatcaggat actatgcggc cgcttacttg
     1801 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     1861 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     1921 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1981 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     2041 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     2101 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     2161 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     2221 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     2281 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     2341 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     2401 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     2461 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catcatatga
     2521 gacatactcg gaattttgtg attccgagca atttgtcttt taactatttc caatgtttta
     2581 ccccaagctt tagggttgtt gaatcttcaa tcccctaagt gttcaagttc aaaaccaagg
     2641 ttaattctca atcttgcttg tttgactgtg aactaatgct gtcctttaac aataatgttt
     2701 ctattaaggt tgttggatca ccctgcagag tctgaggtgg aacttgtgat gtgataagac
     2761 ctaggtgttg acaagggatt ttcaactgag cacactgatg cccttgctcc cttcggagac
     2821 ctaggttgtt ctctatcttt gtgatgtggc tctgaaacaa accaggtgtg attacagtaa
     2881 caatttcaat ttaaattccg atataaattt caatgagagg aaaattatta atcttcctca
     2941 tagttattcg cacacaaaag atcctaaaaa ttcttctttc tttttgtgtg tccggggtcg
     3001 gcatggcatc tccacctcct cgcggtccga cctgggcatc cgaaggagga cgtcgtccac
     3061 tcggatggct aagggagagc tccctagatc ggaactcgct agcagatctt tttccctctg
     3121 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     3181 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     3241 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     3301 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     3361 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     3421 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     3481 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     3541 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     3601 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     3661 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     3721 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     3781 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     3841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     3901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     3961 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     4021 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     4081 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     4141 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4201 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4261 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4321 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4381 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4441 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4501 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4561 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4621 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4681 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4741 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     4801 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4861 gcttgtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4921 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     4981 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5041 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5101 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5161 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5221 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5281 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5341 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5401 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5461 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5521 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5581 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5641 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5701 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5761 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5821 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5881 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     5941 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6001 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6061 acatttcccc gaaaagtgcc acctgggtc
//
