LOCUS       sequence-301586-        5294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Marburg virus minigenome with luciferase reporter gene under
            control of a T7 RNA polymerase promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5294)
  AUTHORS   Manhart WA, Pacheco JR, Hume AJ, Cressey TN, Deflube LR, Muhlberger 
            E
  TITLE     A Chimeric Lloviu Virus Minigenome System Reveals that the 
            Bat-Derived Filovirus Replicates More Similarly to Ebolaviruses than
            Marburgviruses.
  JOURNAL   Cell Rep. 2018 Sep 4;24(10):2573-2580.e4. doi: 
            10.1016/j.celrep.2018.08.008.
  PUBMED    30184492
REFERENCE   2  (bases 1 to 5294)
  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"; doi: 
            "10.1016/j.celrep.2018.08"; journalName: "Cell Rep"; date: 
            "2018-09-4- 4"; volume: "24"; issue: "10"; pages: "2573-2580.e4"
COMMENT     Sequence Label: p2,0_3M5M_luciferase
FEATURES             Location/Qualifiers
     source          1..5294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..42)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(196..784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(276..295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(955..1746,1747..1815))
                     /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     1578..1597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1816..1920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1988..2006
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2044..2066)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2166..2185
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2395..2414
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2704..2722
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2704..2721
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      complement(2790..2837)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2833..2851
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             3116..4768
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(3182..3200)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     promoter        complement(5213..5231)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
       61 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
      121 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
      181 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
      241 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
      301 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
      361 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
      421 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
      481 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
      541 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
      601 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
      661 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
      721 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
      781 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
      841 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
      901 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
      961 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     1021 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     1081 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     1141 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     1201 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     1261 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     1321 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     1381 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     1441 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     1501 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     1561 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     1621 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     1681 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     1741 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     1801 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     1861 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     1921 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     1981 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     2041 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     2101 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     2161 ctatgcggca tcagagcaga ttgtactgag agtgcaccat tcgacgctct cccttatgcg
     2221 actcctgcat taagaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     2281 ggaatggtgc aaggagatgg cgcccaacag tcccccggcc acggggcctg ccaccatacc
     2341 cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc catcggtgat
     2401 gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg
     2461 tccggcgtag aggatctggc tagcgatgac cctgctgatt ggttcgctga ccatttccgg
     2521 gtgcgggacg gcgttaccag aaactcagaa ggttcgtcca accaaaccga ctctgacggc
     2581 agtttacgag agagatgata gggtctgctt cagtaagcca gatgctacac aattaggctt
     2641 gtacatattg tcgttagaac gcggctacaa ttaatacata accttatgta tcatacacat
     2701 acgatttagg tgacactata gaatacaagc ttgcatgcct gcaggtcgac tctagaggat
     2761 ctcgatccgg atatagttcc tcctttcagc aaaaaacccc tcaagacccg tttagaggcc
     2821 ccaaggggtt atgctagtta ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg
     2881 gctttgttag cagccggatc cttttttttg agctctccct tagccatccg agtggacgac
     2941 gtcctccttc ggatgcccag gtcggaccgc gaggaggtgg agatgccatg ccgacccaga
     3001 cacacaaaaa caagagatga tgattttgtg tatcatataa ataaagaaga atattaacat
     3061 tgacattgag acttgtcagt ctgttaatat tcttgaagag atgtttttat ctcatatgga
     3121 agacgccaaa aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc
     3181 tggagagcaa ctgcataagg ctatgaagag atacgccctg gttcctggaa caattgcttt
     3241 tacagatgca catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg
     3301 gttggcagaa gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag
     3361 tgaaaactct cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt
     3421 tgcgcccgcg aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca
     3481 gcctaccgtg gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa
     3541 gctcccaatc atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca
     3601 gtcgatgtac acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt
     3661 gccagagtcc ttcgataggg acaagacaat tgcactgatc atgaactcct ctggatctac
     3721 tggtctgcct aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc
     3781 cagagatcct atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc
     3841 attccatcac ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt
     3901 cgtcttaatg tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat
     3961 tcaaagtgcg ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga
     4021 caaatacgat ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga
     4081 agtcggggaa gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct
     4141 cactgagact acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt
     4201 cggtaaagtt gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct
     4261 gggcgttaat caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt
     4321 aaacaatccg gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga
     4381 catagcttac tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat
     4441 taagtacaaa ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc
     4501 caacatcttc gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc
     4561 cgccgttgtt gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt
     4621 cgccagtcaa gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt
     4681 accgaaaggt cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc
     4741 caagaagggc ggaaagatcg ccgtgtaagc ggccgctagt gattagtgtt tggtcacacc
     4801 agaggtaact ttttaacgtt aattactcag atgttattgc tcataattag cattaatatt
     4861 ggcacattgg gtgaatcctt gagctttatc cctaatatgg tgtaagaaat taaggaaata
     4921 ctgagataca ctagttgaat tgaattatga cataccatat atcataaata taaaaaagtg
     4981 tctgctgtaa tctacaagca cctcttttaa atacattagg aaaagaatta agttaccgtt
     5041 gagatcaaaa aaaccacgtc atgttttctc tgatgacaag tgataaaact tcgtagttaa
     5101 atttctagaa tgtcgatgtg aatgtaaatt aagaaaaacc aatatataaa attaaaaaat
     5161 taaaaagctt tgatataagt aacacaaaac attcttcatc ttttttgtgt gtcctatagt
     5221 gagtcgtatt aatttcgcgg gatcgatatc cccctgcatt aatgaatcgg ccaacgcgcg
     5281 gggagaggcg gttt
//
