LOCUS       sequence-197230-        6563 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Ebola virus VP30 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 6563)
  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 6563)
  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_VP30_EBOV
FEATURES             Location/Qualifiers
     source          1..6563
                     /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(3586..3605)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3651..3706
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3705..3724)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4066..4082)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4066..4082)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4079..4101)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4090..4106
                     /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(4114..4120,4121..4138,4139..4144))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4159..4180
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4239..4434
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4285..4420
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4440..4574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4477..4496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4531..4550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4642..4659)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4813..5401)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4893..4912)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5572..6363,6364..6432))
                     /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     6195..6214
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6433..6537)
                     /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 aagaattccc cggggagctc
     1741 actagtggat ccgcgaaccg gtagagttta gttgcaacct aacacacata aagcattggt
     1801 caaaaagtca atagaaattt aaacagtgag tggagacaac ttttaaatgg aagcttcata
     1861 tgagagagga cgcccacgag ctgccagaca gcattcaagg gatggacacg accaccatgt
     1921 tcgagcacga tcatcatcca gagagaatta tcgaggtgag taccgtcaat caaggagcgc
     1981 ctcacaagtg cgcgttccta ctgtatttca taagaagaga gttgaaccat taacagttcc
     2041 tccagcacct aaagacatat gtccgacctt gaaaaaagga tttttgtgtg acagtagttt
     2101 ttgcaaaaaa gatcaccagt tggagagttt aactgatagg gaattactcc tactaatcgc
     2161 ccgtaagact tgtggatcag tagaacaaca attaaatata actgcaccca aggactcgcg
     2221 cttagcaaat ccaacggctg atgatttcca gcaagaggaa ggtccaaaaa ttaccttgtt
     2281 gacactgatc aagacggcag aacactgggc gagacaagac atcagaacca tagaggattc
     2341 aaaattaaga gcattgttga ctctatgtgc tgtgatgacg aggaaattct caaaatccca
     2401 gctgagtctt ttatgtgaga cacacctaag gcgcgagggg cttgggcaag atcaggcaga
     2461 acccgttctc gaagtatatc aacgattaca cagtgataaa ggaggcagtt ttgaagctgc
     2521 actatggcaa caatgggacc gacaatccct aattatgttt atcactgcat tcttgaatat
     2581 tgctctccag ttaccgtgtg aaagttctgc tgtcgttgtt tcagggttaa gaacattggt
     2641 tcctcaatca gataatgagg aagcttcaac caacccgggg acatgctcat ggtctgatga
     2701 gggtacccct taataaggct gactaaaaca ctatataacc ttctacttga tcacaatact
     2761 ccgtatacct atcatcatat atttaatcaa gacgatatcc tttaaaactt attcagtact
     2821 ataatcactc tcgtttcaaa ttaataagat gtgcatgatt gccctaatat atgaagaggt
     2881 atgatacaac cttaacagtg atcaaagaaa atcataatct cgtatcgctc gtaatataac
     2941 ctgccaagca tacctcttgc acaaagtgat tcttgtacac aaataatgtt ttactctaca
     3001 ggaggtagca acgatccatc ccatcaaaaa ataagtattt catgacttac taatgatctc
     3061 ttaaaatatt aagaaaaact gcggaacata aattctttat gcttcaagct gtggaggagg
     3121 tgtttggtat tggctattgt tatattacaa tcaataacaa gcttgtaaaa atattgttct
     3181 tgtttcaaga ggtagattgt gaccggaaat gctaaactaa tgatgaagat taatgcggag
     3241 gtctgataag aataaacctt attattcaga ttaggcccca agaggcattc ttcatctcct
     3301 tttagcaaag tactatttca gggtagtcca attagtggca cgtcttttag ctgtatatca
     3361 gtcgcccctg agatacgcca caaaagtgtc tctaagctaa attggtctgt acacatccca
     3421 tacattgtat taggggcaat aatatctaat tgaacttagc cgtttaaaat ttagtgcata
     3481 aatctgggct aacaccacca ggtcaactcc attggctgaa aagaagctta cctgcagctc
     3541 gagggtaccc ccggggcggc cgcgagctct taattagcta gcagatcttt ttccctctgc
     3601 caaaaattat ggggacatca tgaagcccct tgagcatctg acttctggct aataaaggaa
     3661 atttattttc attgcaatag tgtgttggaa ttttttgtgt ctctcactcg gaaggacata
     3721 tgggagggca aatcatttaa aacatcagaa tgagtatttg gtttagagtt tggcaacata
     3781 tgcccatatg ctggctgcca tgaacaaagg tggctataaa gaggtcatca gtatatgaaa
     3841 cagccccctg ctgtccattc cttattccat agaaaagcct tgacttgagg ttagattttt
     3901 tttatatttt gttttgtgtt atttttttct ttaacatccc taaaattttc cttacatgtt
     3961 ttactagcca gatttttcct cctctcctga ctactcccag tcatagctgt ccctcttctc
     4021 ttatgaagat ccctcgacct gcagcccaag cttggcgtaa tcatggtcat agctgtttcc
     4081 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4141 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4201 cgctttccag tcgggaaacc tgtcgtgcca gcggatccgc atctcaatta gtcagcaacc
     4261 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     4321 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     4381 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagcta
     4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4561 atcatgtctg gatccgctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     4621 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4681 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4741 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     4801 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     4861 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     4921 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     4981 cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca gttcggtgta
     5041 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5101 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5161 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5221 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5281 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5341 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5401 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5461 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     5521 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     5581 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     5641 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     5701 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     5761 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     5821 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     5881 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     5941 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     6001 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     6061 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     6121 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     6181 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     6241 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     6301 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     6361 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     6421 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     6481 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     6541 atttccccga aaagtgccac ctg
//
