LOCUS       Exported                6744 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encodes for Ebola Virus Zaire VP35 under control of a T7 RNA 
            polymerase promoter..
ACCESSION   .
VERSION     .
KEYWORDS    pTM1_VP35_ZEBOV
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6744)
  AUTHORS   Trunschke M, Conrad D, Enterlein S, Olejnik J, Brauburger K, 
            Muhlberger E
  TITLE     The L-VP35 and L-L interaction domains reside in the amino terminus 
            of the Ebola virus L protein and are potential targets for 
            antivirals.
  JOURNAL   Virology. 2013 Jul 5;441(2):135-45. doi: 
            10.1016/j.virol.2013.03.013. Epub 2013 Apr 11.
  PUBMED    23582637
REFERENCE   2  (bases 1 to 6744)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6744
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     794..813
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        794..812
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    851..1423
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1018..1035)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1245..1264
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(2929..2947)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2943..2990
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(3970..3992)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      4122..4577
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4209..4228)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4419..4440
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4606..4710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4711..5571
                     /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(4929..4948)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5742..6330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6231..6250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6484..6501
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6618..6639
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        6654..6684
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6692..6708
                     /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)."
     primer_bind     6697..6719
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6716..6732
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6716..6732
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttttgc gatcaataaa tggatcacaa ccagtatctc ttaacgatgt tcttcgcaga
       61 tgatgattca ttttttaagt atttggctag tcaagatgat gaatcttcat tatctgatat
      121 attgcaaatc actcaatatc tagactttct gttattatta ttgatccaat caaaaaataa
      181 attagaagcc gtgggtcatt gttatgaatc tctttcagag gaatacagac aattgacaaa
      241 attcacagac tttcaagatt ttaaaaaact gtttaacaag gtccctattg ttacagatgg
      301 aagggtcaaa cttaataaag gatatttgtt cgactttgtg attagtttga tgcgattcaa
      361 aaaagaatcc tctctagcta ccaccgcaat agatcctgtt agatacatag atcctcgtcg
      421 caatatcgca ttttctaacg tgatggatat attaaagtcg aataaagtga acaataatta
      481 attctttatt gtcatcatga acggcggaca tattcagttg ataatcggcc ccatgttttc
      541 aggtaaaagt acagaattaa ttagacgagt tagacgttat caaatagctc aatataaatg
      601 cgtgactata aaatattcta acgataatag atacggaacg ggactatgga cgcatgataa
      661 gaataatttt gaagcattgg aagcaactaa actatgtgat gtcttggaat caattacaga
      721 tttctccgtg ataggtatcg atgaaggaca gttctttcca gacattgttg aattgatctc
      781 gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc ctctagcggg
      841 atcaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
      901 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
      961 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1021 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     1081 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1141 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1201 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     1261 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     1321 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaaacgt ctaggccccc
     1381 cgaaccacgg ggacgtggtt ttcctttgaa aaacacgata ataccatggg aattccccgg
     1441 ggagctcact agtggatcca aaccttcatc atccttacgt caattgaatt ctctagcact
     1501 cgaagcttat tgtcttcaat gtaaaagaaa agctggtcta acaagatgac aactagaaca
     1561 aagggcaggg gccatactgc ggccacgact caaaacgaca gaatgccagg ccctgagctt
     1621 tcgggctgga tctctgagca gctaatgacc ggaagaattc ctgtaagcga catcttctgt
     1681 gatattgaga acaatccagg attatgctac gcatcccaaa tgcaacaaac gaagccaaac
     1741 ccgaagacgc gcaacagtca aacccaaacg gacccaattt gcaatcatag ttttgaggag
     1801 gtagtacaaa cattggcttc attggctact gttgtgcaac aacaaaccat cgcatcagaa
     1861 tcattagaac aacgcattac gagtcttgag aatggtctaa agccagttta tgatatggca
     1921 aaaacaatct cctcattgaa cagggtttgt gctgagatgg ttgcaaaata tgatcttctg
     1981 gtgatgacaa ccggtcgggc aacagcaacc gctgcggcaa ctgaggctta ttgggccgaa
     2041 catggtcaac caccacctgg accatcactt tatgaagaaa gtgcgattcg gggtaagatt
     2101 gaatctagag atgagaccgt ccctcaaagt gttagggagg cattcaacaa tctaaacagt
     2161 accacttcac taactgagga aaattttggg aaacctgaca tttcggcaaa ggatttgaga
     2221 aacattatgt atgatcactt gcctggtttt ggaactgctt tccaccaatt agtacaagtg
     2281 atttgtaaat tgggaaaaga tagcaactca ttggacatca ttcatgctga gttccaggcc
     2341 agcctggctg aaggagactc tcctcaatgt gccctaattc aaattacaaa aagagttcca
     2401 atcttccaag atgctgctcc acctgtcatc cacatccgct ctcgaggtga cattccccga
     2461 gcttgccaga aaagcttgcg tccagtccca ccatcgccca agattgatcg aggttgggta
     2521 tgtgtttttc agcttcaaga tggtaaaaca cttggactca aaatttgagc caatctccct
     2581 tccctccgaa agaggcgaat aatagcagag gcttcaactg ctgaactata gggtacgtta
     2641 cattaatgat acacttgtga gtatcagccc tggataatat aagtcaatta aacgaccaag
     2701 ataaaattgt tcatatctcg ctagcagctt aaaatataaa tgtaatagga gctatatctc
     2761 tgacagtatt ataatcaatt gttattaagt aacccaaacc aaaagtgatg aagattaaga
     2821 aaaacctacc tcggctgaga gagtgttttt tcattaacct tcatcttgta aacgtcgacg
     2881 atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat
     2941 aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag
     3001 gaactatatc cggatcgaga tcaattctgt gagcgtatgg caaacgaagg aaaaatagtt
     3061 atagtagccg cactcgatgg gacatttcaa cgtaaaccgt ttaataatat tttgaatctt
     3121 attccattat ctgaaatggt ggtaaaacta actgctgtgt gtatgaaatg ctttaaggag
     3181 gcttcctttt ctaaacgatt gggtgaggaa accgagatag aaataatagg aggtaatgat
     3241 atgtatcaat cggtgtgtag aaagtgttac atcgactcat aatattatat tttttatcta
     3301 aaaaactaaa aataaacatt gattaaattt taatataata cttaaaaatg gatgttgtgt
     3361 cgttagataa accgtttatg tattttgagg aaattgataa tgagttagat tacgaaccag
     3421 aaagtgcaaa tgaggtcgca aaaaaactgc cgtatcaagg acagttaaaa ctattactag
     3481 gagaattatt ttttcttagt aagttacagc gacacggtat attagatggt gccaccgtag
     3541 tgtatatagg atctgctccc ggtacacata tacgttattt gagagatcat ttctataatt
     3601 taggagtgat catcaaatgg atgctaattg acggccgcca tcatgatcct attttaaatg
     3661 gattgcgtga tgtgactcta gtgactcggt tcgttgatga ggaatatcta cgatccatca
     3721 aaaaacaact gcatccttct aagattattt taatttctga tgtgagatcc aaacgaggag
     3781 gaaatgaacc tagtacggcg gatttactaa gtaattacgc tctacaaaat gtcatgatta
     3841 gtattttaaa ccccgtggcg tctagtctta aatggagatg cccgtttcca gatcaatgga
     3901 tcaaggactt ttatatccca cacggtaata aaatgttaca accttttgct ccttcatatt
     3961 cagggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     4021 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     4081 cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc gacgcgccct gtagcggcgc
     4141 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     4201 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     4261 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     4321 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     4381 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     4441 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     4501 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     4561 attaacgttt acaatttccc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     4621 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     4681 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4741 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4801 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4861 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4921 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4981 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     5041 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     5101 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     5161 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     5221 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     5281 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     5341 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     5401 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     5461 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     5521 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     5581 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     5641 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     5701 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5761 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5821 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5881 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5941 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     6001 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     6061 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     6121 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     6181 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     6241 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     6301 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     6361 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     6421 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     6481 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     6541 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     6601 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     6661 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6721 aacagctatg accatgatta cgcc
//
