LOCUS       sequence-197128-        6855 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Ebola virus minigenome with EGFP 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 6855)
  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 6855)
  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..6855
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1674
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1598..1620
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1682..1701
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             complement(join(2565..3278,3279..3281,3282..3284))
                     /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(2568..3278,3279..3281,3282..3284))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2610..2631)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     2959..2978
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3218..3239
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3878..3897)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3943..3998
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3997..4016)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4358..4374)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4358..4374)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4371..4393)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4382..4398
                     /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(4406..4412,4413..4430,4431..4436))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4451..4472
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4531..4726
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4577..4712
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4732..4866
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4769..4788)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4823..4842
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4934..4951)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5105..5693)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5185..5204)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5864..6655,6656..6724))
                     /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     6487..6506
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6725..6829)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcaag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcagctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattcct cgagggtacc
     1741 cccggggcgg ccgcgagctc ttaattagct agcgaattgt gtgtccactg atgagtccgt
     1801 gaggacgaaa cccggagtcc cgggtctgga cacacaaaaa agaagaaata gatttatttt
     1861 taaatttttg tgtgcgacca tttttccagg aatccttttt gcaacgttta ttctggcgag
     1921 ccttctgatg agcgtggtca atgtcttaat tatcattaac acgaagatta ttatctagtc
     1981 tgctatattg taaagaaggt atttctgatt ttacagtcct gccttttctt ttaattttat
     2041 cattctgtat gaaatagtag cttactagac aaaaatgtta aggttattac acctaatctc
     2101 ttattttaca caaagattaa ggctatcacc gtttttaaaa cgacaaatta aagtaatgat
     2161 aattataaaa aacttaatct taaagatcaa ttttgtttta agaagttgtg ttatcaacca
     2221 aaacactatt ccatctgaca ggatattgat acaacattat tatatcaata acaatatgag
     2281 cccagacctt tcgttaaagc ttaattataa agagtgcaga gtttattatg ttgcgttaaa
     2341 ttcattgcgg cagtaatatg tcacacatgc tgcattgtga attttttgat tatcacgcag
     2401 ggagagaggc taaatatagt atataatata gggactcctt tcgttattta aatagttgtt
     2461 tattgagatt agatcaatat gatgtatgag agcaatttat gagtttttta taatctgtat
     2521 gttaataacc aacctttgca agtatcagga tactatgcgg ccgcttactt gtacagctcg
     2581 tccatgccga gagtgatccc ggcggcggtc acgaactcca gcaggaccat gtgatcgcgc
     2641 ttctcgttgg ggtctttgct cagggcggac tgggtgctca ggtagtggtt gtcgggcagc
     2701 agcacggggc cgtcgccgat gggggtgttc tgctggtagt ggtcggcgag ctgcacgctg
     2761 ccgtcctcga tgttgtggcg gatcttgaag ttcaccttga tgccgttctt ctgcttgtcg
     2821 gccatgatat agacgttgtg gctgttgtag ttgtactcca gcttgtgccc caggatgttg
     2881 ccgtcctcct tgaagtcgat gcccttcagc tcgatgcggt tcaccagggt gtcgccctcg
     2941 aacttcacct cggcgcgggt cttgtagttg ccgtcgtcct tgaagaagat ggtgcgctcc
     3001 tggacgtagc cttcgggcat ggcggacttg aagaagtcgt gctgcttcat gtggtcgggg
     3061 tagcggctga agcactgcac gccgtaggtc agggtggtca cgagggtggg ccagggcacg
     3121 ggcagcttgc cggtggtgca gatgaacttc agggtcagct tgccgtaggt ggcatcgccc
     3181 tcgccctcgc cggacacgct gaacttgtgg ccgtttacgt cgccgtccag ctcgaccagg
     3241 atgggcacca ccccggtgaa cagctcctcg cccttgctca ccatcatatg agacatactc
     3301 ggaattttgt gattccgagc aatttgtctt ttaactattt ccaatgtttt accccaagct
     3361 ttagggttgt tgaatcttca atcccctaag tgttcaagtt caaaaccaag gttaattctc
     3421 aatcttgctt gtttgactgt gaactaatgc tgtcctttaa caataatgtt tctattaagg
     3481 ttgttggatc accctgcaga gtctgaggtg gaacttgtga tgtgataaga cctaggtgtt
     3541 gacaagggat tttcaactga gcacactgat gcccttgctc ccttcggaga cctaggttgt
     3601 tctctatctt tgtgatgtgg ctctgaaaca aaccaggtgt gattacagta acaatttcaa
     3661 tttaaattcc gatataaatt tcaatgagag gaaaattatt aatcttcctc atagttattc
     3721 gcacacaaaa gatcctaaaa attcttcttt ctttttgtgt gtccggggtc ggcatggcat
     3781 ctccacctcc tcgcggtccg acctgggcat ccgaaggagg acgtcgtcca ctcggatggc
     3841 taagggagag ctccctagat cggaactcgc tagcagatct ttttccctct gccaaaaatt
     3901 atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt
     3961 tcattgcaat agtgtgttgg aattttttgt gtctctcact cggaaggaca tatgggaggg
     4021 caaatcattt aaaacatcag aatgagtatt tggtttagag tttggcaaca tatgcccata
     4081 tgctggctgc catgaacaaa ggtggctata aagaggtcat cagtatatga aacagccccc
     4141 tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt tttttatatt
     4201 ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg ttttactagc
     4261 cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc tcttatgaag
     4321 atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa
     4381 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
     4441 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     4501 agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa ccatagtccc
     4561 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4621 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4681 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc taacttgttt
     4741 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     4801 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     4861 tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     4921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     4981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     5041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     5101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     5161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     5221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     5281 gaagcgtggc gctttctcaa tgctcacgct gtaggtatct cagttcggtg taggtcgttc
     5341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     5401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     5461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     5521 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     5581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     5641 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     5701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     5761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     5821 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     5881 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     5941 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6001 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6061 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6121 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6181 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6241 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     6301 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     6361 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     6421 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     6481 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     6541 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     6601 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     6661 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     6721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     6781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     6841 gaaaagtgcc acctg
//
