LOCUS       sequence-197235-        6011 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Ebola virus VP35 gene (Zaire ebolavirus, Mayinga) under 
            control of the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6011)
  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 6011)
  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_VP35_EBOV
FEATURES             Location/Qualifiers
     source          1..6011
                     /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"
     primer_bind     complement(3034..3053)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3099..3154
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3153..3172)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3514..3530)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3514..3530)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3527..3549)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3538..3554
                     /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(3562..3568,3569..3586,3587..3592))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3607..3628
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3687..3882
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3733..3868
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3888..4022
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3925..3944)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3979..3998
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4090..4107)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4261..4849)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4341..4360)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5020..5811,5812..5880))
                     /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     5643..5662
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5881..5985)
                     /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 cccggggagc tcactagtgg atccaaacct tcatcatcct tacgtcaatt gaattctcta
     1801 gcactcgaag cttattgtct tcaatgtaaa agaaaagctg gtctaacaag atgacaacta
     1861 gaacaaaggg caggggccat actgcggcca cgactcaaaa cgacagaatg ccaggccctg
     1921 agctttcggg ctggatctct gagcagctaa tgaccggaag aattcctgta agcgacatct
     1981 tctgtgatat tgagaacaat ccaggattat gctacgcatc ccaaatgcaa caaacgaagc
     2041 caaacccgaa gacgcgcaac agtcaaaccc aaacggaccc aatttgcaat catagttttg
     2101 aggaggtagt acaaacattg gcttcattgg ctactgttgt gcaacaacaa accatcgcat
     2161 cagaatcatt agaacaacgc attacgagtc ttgagaatgg tctaaagcca gtttatgata
     2221 tggcaaaaac aatctcctca ttgaacaggg tttgtgctga gatggttgca aaatatgatc
     2281 ttctggtgat gacaaccggt cgggcaacag caaccgctgc ggcaactgag gcttattggg
     2341 ccgaacatgg tcaaccacca cctggaccat cactttatga agaaagtgcg attcggggta
     2401 agattgaatc tagagatgag accgtccctc aaagtgttag ggaggcattc aacaatctaa
     2461 acagtaccac ttcactaact gaggaaaatt ttgggaaacc tgacatttcg gcaaaggatt
     2521 tgagaaacat tatgtatgat cacttgcctg gttttggaac tgctttccac caattagtac
     2581 aagtgatttg taaattggga aaagatagca actcattgga catcattcat gctgagttcc
     2641 aggccagcct ggctgaagga gactctcctc aatgtgccct aattcaaatt acaaaaagag
     2701 ttccaatctt ccaagatgct gctccacctg tcatccacat ccgctctcga ggtgacattc
     2761 cccgagcttg ccagaaaagc ttgcgtccag tcccaccatc gcccaagatt gatcgaggtt
     2821 gggtatgtgt ttttcagctt caagatggta aaacacttgg actcaaaatt tgagccaatc
     2881 tcccttccct ccgaaagagg cgaataatag cagaggcttc aactgctgaa ctatagggta
     2941 cgttacatta atgatacact tgtgagtatc agccctggat aatataagtc aattaaacga
     3001 ccaagataaa attgttcata tctcgctagc agatcttttt ccctctgcca aaaattatgg
     3061 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     3121 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa
     3181 tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct
     3241 ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     3301 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     3361 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     3421 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atgaagatcc
     3481 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     3541 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     3601 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     3661 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     3721 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3781 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     3841 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     3901 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     3961 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     4021 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4081 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4141 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4201 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4261 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     4321 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     4381 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     4441 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     4501 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     4561 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     4621 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     4681 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     4741 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     4801 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     4861 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     4921 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     4981 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5041 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5101 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5161 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5221 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     5281 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     5341 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     5401 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     5461 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     5521 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     5581 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     5641 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     5701 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     5761 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     5821 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     5881 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     5941 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6001 agtgccacct g
//
