LOCUS       Exported                5016 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encodes for Ebola Minigenome with eGFP reporter gene under control 
            of a T7 RNA polymerase promoter.
ACCESSION   .
VERSION     .
KEYWORDS    p2,0_3E5E_eGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5016)
  AUTHORS   Schmidt KM, Schumann M, Olejnik J, Krahling V, Muhlberger E
  TITLE     Recombinant Marburg virus expressing EGFP allows rapid screening of 
            virus growth and real-time visualization of virus spread.
  JOURNAL   J Infect Dis. 2011 Nov;204 Suppl 3:S861-70. doi: 
            10.1093/infdis/jir308.
  PUBMED    21987762
REFERENCE   2  (bases 1 to 5016)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5016
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             482..1201
                     /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             482..1198
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(527..548)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(788..807)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1135..1156
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1940..1957)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2042..2059)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2213..2801)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2293..2312)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2972..3832)
                     /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     3595..3614
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3833..3937)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4005..4023
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(4061..4083)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4183..4202
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4413..4432
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4723..4741
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     4723..4740
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     terminator      4809..4856
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4852..4870
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
ORIGIN
        1 cggacacaca aaaagaaaga agaattttta ggatcttttg tgtgcgaata actatgagga
       61 agattaataa ttttcctctc attgaaattt atatcggaat ttaaattgaa attgttactg
      121 taatcacacc tggtttgttt cagagccaca tcacaaagat agagaacaac ctaggtctcc
      181 gaagggagca agggcatcag tgtgctcagt tgaaaatccc ttgtcaacac ctaggtctta
      241 tcacatcaca agttccacct cagactctgc agggtgatcc aacaacctta atagaaacat
      301 tattgttaaa ggacagcatt agttcacagt caaacaagca agattgagaa ttaaccttgg
      361 ttttgaactt gaacacttag gggattgaag attcaacaac cctaaagctt ggggtaaaac
      421 attggaaata gttaaaagac aaattgctcg gaatcacaaa attccgagta tgtctcatat
      481 gatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
      541 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
      601 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
      661 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
      721 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
      781 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
      841 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
      901 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
      961 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1021 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     1081 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     1141 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     1201 agcggccgca tagtatcctg atacttgcaa aggttggtta ttaacataca gattataaaa
     1261 aactcataaa ttgctctcat acatcatatt gatctaatct caataaacaa ctatttaaat
     1321 aacgaaagga gtccctatat tatatactat atttagcctc tctccctgcg tgataatcaa
     1381 aaaattcaca atgcagcatg tgtgacatat tactgccgca atgaatttaa cgcaacataa
     1441 taaactctgc actctttata attaagcttt aacgaaaggt ctgggctcat attgttattg
     1501 atataataat gttgtatcaa tatcctgtca gatggaatag tgttttggtt gataacacaa
     1561 cttcttaaaa caaaattgat ctttaagatt aagtttttta taattatcat tactttaatt
     1621 tgtcgtttta aaaacggtga tagccttaat ctttgtgtaa aataagagat taggtgtaat
     1681 aaccttaaca tttttgtcta gtaagctact atttcataca gaatgataaa attaaaagaa
     1741 aaggcaggac tgtaaaatca gaaatacctt ctttacaata tagcagacta gataataatc
     1801 ttcgtgttaa tgataattaa gacattgacc acgctcatca gaaggctcgc cagaataaac
     1861 gttgcaaaaa ggattcctgg aaaaatggtc gcacacaaaa atttaaaaat aaatctattt
     1921 cttctttttt gtgtgtccac tatagtgagt cgtattaacc cgggatcgat atccccctgc
     1981 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     2041 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     2101 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     2161 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     2221 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     2281 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     2341 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     2401 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     2461 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     2521 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     2581 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     2641 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     2701 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     2761 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     2821 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     2881 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     2941 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     3001 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     3061 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     3121 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     3181 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     3241 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     3301 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     3361 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     3421 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     3481 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     3541 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     3601 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     3661 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     3721 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     3781 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     3841 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     3901 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     3961 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     4021 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     4081 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     4141 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     4201 tactgagagt gcaccatatc gacgctctcc cttatgcgac tcctgcatta agaagcagcc
     4261 cagtagtagg ttgaggccgt tgagcaccgc cgccgcaagg aatggtgcaa ggagatggcg
     4321 cccaacagtc ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg
     4381 agcccgaagt ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca
     4441 accgcacctg tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatctggcta
     4501 gcgatgaccc tgctgattgg ttcgctgacc atttccgggg tgcggaacgg cgttaccaga
     4561 aactcagaag gttcgtccaa ccaaaccgac tctgacggca gtttacgaga gagatgatag
     4621 ggtctgcttc agtaagccag atgctacaca attaggcttg tacatattgt cgttagaacg
     4681 cggctacaat taatacataa ccttatgtat catacacata cgatttaggt gacactatag
     4741 aatacaagct tgcatgcctg caggtcgact ctagaggatc tcgatccgga tatagttcct
     4801 cctttcagca aaaaacccct caagacccgt ttagaggccc caaggggtta tgctagttat
     4861 tgctcagcgg tggcagcagc caactcagct tcctttcggg ctttgttagc agccggatcc
     4921 ttttttttga gctctccctt agccatccga gtggacgacg tcctccttcg gatgcccagg
     4981 tcggaccgcg aggaggtgga gatgccatgc cgaccc
//
