LOCUS       sequence-268921-        4364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes Marburg virus minigenome with eGFP 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 4364)
  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 4364)
  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_eGFP
FEATURES             Location/Qualifiers
     source          1..4364
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     386..403
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        491..509
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             complement(join(954..1667,1668..1670,1671..1673))
                     /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(957..1667,1668..1670,1671..1673))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(999..1020)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1348..1367
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1607..1628
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1941..1959)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1955..2002
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(2070..2088)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2071..2088)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2378..2397)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     complement(2607..2626)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2726..2748
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2786..2804)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2872..2976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2977..3045,3046..3837)
                     /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     complement(3195..3214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(4008..4364,1..232)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
       61 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
      121 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
      181 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
      241 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
      301 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
      361 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
      421 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcaggg ggatatcgat
      481 cccgcgaaat taatacgact cactatagga cacacaaaaa agatgaagaa tgttttgtgt
      541 tacttatatc aaagcttttt aattttttaa ttttatatat tggtttttct taatttacat
      601 tcacatcgac attctagaaa tttaactacg aagttttatc acttgtcatc agagaaaaca
      661 tgacgtggtt tttttgatct caacggtaac ttaattcttt tcctaatgta tttaaaagag
      721 gtgcttgtag attacagcag acactttttt atatttatga tatatggtat gtcataattc
      781 aattcaacta gtgtatctca gtatttcctt aatttcttac accatattag ggataaagct
      841 caaggattca cccaatgtgc caatattaat gctaattatg agcaataaca tctgagtaat
      901 taacgttaaa aagttacctc tggtgtgacc aaacactaat cactagcggc cgcttacttg
      961 tacagctcgt ccatgccgag agtgatcccg gcggcggtca cgaactccag caggaccatg
     1021 tgatcgcgct tctcgttggg gtctttgctc agggcggact gggtgctcag gtagtggttg
     1081 tcgggcagca gcacggggcc gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc
     1141 tgcacgctgc cgtcctcgat gttgtggcgg atcttgaagt tcaccttgat gccgttcttc
     1201 tgcttgtcgg ccatgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc
     1261 aggatgttgc cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg
     1321 tcgccctcga acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg
     1381 gtgcgctcct ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg
     1441 tggtcggggt agcggctgaa gcactgcacg ccgtaggtca gggtggtcac gagggtgggc
     1501 cagggcacgg gcagcttgcc ggtggtgcag atgaacttca gggtcagctt gccgtaggtg
     1561 gcatcgccct cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc
     1621 tcgaccagga tgggcaccac cccggtgaac agctcctcgc ccttgctcac catcatatga
     1681 gataaaaaca tctcttcaag aatattaaca gactgacaag tctcaatgtc aatgttaata
     1741 ttcttcttta tttatatgat acacaaaatc atcatctctt gtttttgtgt gtctgggtcg
     1801 gcatggcatc tccacctcct cgcggtccga cctgggcatc cgaaggagga cgtcgtccac
     1861 tcggatggct aagggagagc tcaaaaaaaa ggatccggct gctaacaaag cccgaaagga
     1921 agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa
     1981 acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatcga gatcctctag
     2041 agtcgacctg caggcatgca agcttgtatt ctatagtgtc acctaaatcg tatgtgtatg
     2101 atacataagg ttatgtatta attgtagccg cgttctaacg acaatatgta caagcctaat
     2161 tgtgtagcat ctggcttact gaagcagacc ctatcatctc tctcgtaaac tgccgtcaga
     2221 gtcggtttgg ttggacgaac cttctgagtt tctggtaacg ccgtcccgca cccggaaatg
     2281 gtcagcgaac caatcagcag ggtcatcgct agccagatcc tctacgccgg acgcatcgtg
     2341 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     2401 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     2461 gcaggccccg tggccggggg actgttgggc gccatctcct tgcaccattc cttgcggcgg
     2521 cggtgctcaa cggcctcaac ctactactgg gctgcttctt aatgcaggag tcgcataagg
     2581 gagagcgtcg aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     2641 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     2701 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     2761 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     2821 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     2881 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     2941 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     3001 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     3061 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     3121 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     3181 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     3241 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     3301 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     3361 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     3421 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     3481 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     3541 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     3601 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     3661 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     3721 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     3781 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     3841 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     3901 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     3961 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     4021 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     4081 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     4141 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     4201 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     4261 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     4321 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaac
//
