LOCUS       sequence-197238-        7132 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Ebola virus NP 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 7132)
  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 7132)
  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_NP_EBOV
FEATURES             Location/Qualifiers
     source          1..7132
                     /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             3335..3346
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4155..4174)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4220..4275
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4274..4293)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4635..4651)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4635..4651)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4648..4670)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4659..4675
                     /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(4683..4689,4690..4707,4708..4713))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4728..4749
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4808..5003
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4854..4989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5009..5143
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5046..5065)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5100..5119
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5211..5228)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5382..5970)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5462..5481)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6141..6932,6933..7001))
                     /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     6764..6783
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7002..7106)
                     /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 actagtggat ccacaaaatt ccgagtatgg attctcgtcc
     1801 tcagaaaatc tggatggcgc cgagtctcac tgaatctgac atggattacc acaagatctt
     1861 gacagcaggt ctgtccgttc aacaggggat tgttcggcaa agagtcatcc cagtgtatca
     1921 agtaaacaat cttgaagaaa tttgccaact tatcatacag gcctttgaag caggtgttga
     1981 ttttcaagag agtgcggaca gtttccttct catgctttgt cttcatcatg cgtaccaggg
     2041 agattacaaa cttttcttgg aaagtggcgc agtcaagtat ttggaagggc acgggttccg
     2101 ttttgaagtc aagaagcgtg atggagtgaa gcgccttgag gaattgctgc cagcagtatc
     2161 tagtggaaaa aacattaaga gaacacttgc tgccatgccg gaagaggaga caactgaagc
     2221 taatgccggt cagtttctct ccttcgcaag tctattcctt ccgaaattgg tagtaggaga
     2281 aaaggcttgc cttgagaagg ttcaaaggca aattcaagta catgcagagc aaggactgat
     2341 acaatatcca acagcttggc aatcagtagg acacatgatg gtgattttcc gtttgatgcg
     2401 aacaaatttt ctgatcaaat ttctcctaat acaccaaggg atgcacatgg ttgccgggca
     2461 tgatgccaac gatgctgtga tttcaaattc agtggctcaa gctcgttttt caggcttatt
     2521 gattgtcaaa acagtacttg atcatatcct acaaaagaca gaacgaggag ttcgtctcca
     2581 tcctcttgca aggaccgcca aggtaaaaaa tgaggtgaac tcctttaagg ctgcactcag
     2641 ctccctggcc aagcatggag agtatgctcc tttcgcccga cttttgaacc tttctggagt
     2701 aaataatctt gagcatggtc ttttccctca actatcggca attgcactcg gagtcgccac
     2761 agcacacggg agtaccctcg caggagtaaa tgttggagaa cagtatcaac aactcagaga
     2821 ggctgccact gaggctgaga agcaactcca acaatatgca gagtctcgcg aacttgacca
     2881 tcttggactt gatgatcagg aaaagaaaat tcttatgaac ttccatcaga aaaagaacga
     2941 aatcagcttc cagcaaacaa acgctatggt aactctaaga aaagagcgcc tggccaagct
     3001 gacagaagct atcactgctg cgtcactgcc caaaacaagt ggacattacg atgatgatga
     3061 cgacattccc tttccaggac ccatcaatga tgacgacaat cctggccatc aagatgatga
     3121 tccgactgac tcacaggata cgaccattcc cgatgtggtg gttgatcccg atgatggaag
     3181 ctacggcgaa taccagagtt actcggaaaa cggcatgaat gcaccagatg acttggtcct
     3241 attcgatcta gacgaggacg acgaggacac taagccagtg cctaatagat cgaccaaggg
     3301 tggacaacag aagaacagtc aaaagggcca gcatatagag ggcagacaga cacaatccag
     3361 gccaattcaa aatgtcccag gccctcacag aacaatccac cacgccagtg cgccactcac
     3421 ggacaatgac agaagaaatg aaccctccgg ctcaaccagc cctcgcatgc tgacaccaat
     3481 taacgaagag gcagacccac tggacgatgc cgacgacgag acgtctagcc ttccgccctt
     3541 ggagtcagat gatgaagagc aggacaggga cggaacttcc aaccgcacac ccactgtcgc
     3601 cccaccggct cccgtataca gagatcactc tgaaaagaaa gaactcccgc aagacgagca
     3661 acaagatcag gaccacactc aagaggccag gaaccaggac agtgacaaca cccagtcaga
     3721 acactctttt gaggagatgt atcgccacat tctaagatca caggggccat ttgatgctgt
     3781 tttgtattat catatgatga aggatgagcc tgtagttttc agtaccagtg atggcaaaga
     3841 gtacacgtat ccagactccc ttgaagagga atatccacca tggctcactg aaaaagaggc
     3901 tatgaatgaa gagaatagat ttgttacatt ggatggtcaa caattttatt ggccggtgat
     3961 gaatcacaag aataaattca tggcaatcct gcaacatcat cagtgaatga gcatggaaca
     4021 atgggatgat tcaaccgaca aatagctaac attaagtagt caaggaacga aaacaggaag
     4081 aatttttgat gtctaaggtg tgaattatta tcacaataaa agtgattctt atttttgaat
     4141 ttaaagctag cagatctttt tccctctgcc aaaaattatg gggacatcat gaagcccctt
     4201 gagcatctga cttctggcta ataaaggaaa tttattttca ttgcaatagt gtgttggaat
     4261 tttttgtgtc tctcactcgg aaggacatat gggagggcaa atcatttaaa acatcagaat
     4321 gagtatttgg tttagagttt ggcaacatat gcccatatgc tggctgccat gaacaaaggt
     4381 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     4441 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     4501 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     4561 tactcccagt catagctgtc cctcttctct tatgaagatc cctcgacctg cagcccaagc
     4621 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4681 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4741 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4801 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4861 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4921 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4981 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     5041 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5101 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     5161 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5221 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5281 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5341 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5401 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5461 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5521 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcaatgc
     5581 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5641 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5701 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     5761 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     5821 aggacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     5881 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     5941 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6001 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6061 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6121 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6181 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6241 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6301 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6361 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6421 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6481 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6541 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6601 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6661 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6721 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     6781 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     6841 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     6901 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     6961 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7021 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7081 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tg
//
