LOCUS       sequence-234622-        8223 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ectopic expression of ZIKV protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8223)
  AUTHORS   Hertzog J, Dias Junior AG, Rigby RE, Donald CL, Mayer A, Sezgin E, 
            Song C, Jin B, Hublitz P, Eggeling C, Kohl A, Rehwinkel J
  TITLE     Infection with a Brazilian isolate of Zika virus generates RIG-I 
            stimulatory RNA and the viral NS5 protein blocks type I IFN 
            induction and signaling.
  JOURNAL   Eur J Immunol. 2018 Mar 23. doi: 10.1002/eji.201847483.
  PUBMED    29572905
REFERENCE   2  (bases 1 to 8223)
  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: "Eur J 
            Immunol. 2018 Mar 23. doi: 10.1002/eji.201847483."
COMMENT     Sequence Label: pcDNA3.2/NS5-Mt-mut
FEATURES             Location/Qualifiers
     source          1..8223
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    297..321
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1147..1158)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3049..3073)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3099..3140
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3180..3199)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3224..3272
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3474..3902
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3561..3580)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3771..3792
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3911..3931)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3916..4245
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4096..4231
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4158..4177
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4312..5106
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4366..4385)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4976..4995
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5282..5403
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5319..5338)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5373..5392
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5452..5468)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5452..5468)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5465..5487)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5476..5492
                     /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(5500..5506,5507..5524,5525..5530))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5545..5566
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5683..5700)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5854..6442)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5934..5953)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6613..7404,7405..7473))
                     /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     7236..7255
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7474..7578)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7653..7672)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7844..8223
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagttaag ctatcaacaa
      301 gtttgtacaa aaaagcaggc ttcaggagga cagctatggg gggtggaaca ggagagaccc
      361 tgggagagaa atggaaggcc cgcttgaacc agatgtcggc cctggagttc tactcctaca
      421 aaaagtcagg catcaccgag gtgtgcagag aagaggcccg ccgcgccctc aaggatggtg
      481 tggcaacggg aggccatgct gtgtcccgag gaagtgcaaa gctgagatgg ttggtggagc
      541 ggggatacct gcagccctat ggaaaggtca ttgatcttgg atgtggcaga gggggctgga
      601 gttactacgc cgccaccatc cgcaaagttc aagaagtgaa aggatacaca aaaggaggcc
      661 ctggtcatga agaacccgtg ttggtgcaaa gctatgggtg gaacatagtc cgtcttaaga
      721 gtggggtgga cgtctttcat atggcggctg agccgtgtga cacgttgctg tgtgacatag
      781 gtgagtcatc atctagtcct gaagtggaag aagcacggac gctcagagtc ctctccatgg
      841 tgggggattg gcttgaaaaa agaccaggag ccttttgtat aaaagtgttg tgcccataca
      901 ccagcactat gatggaaacc ctggagcgac tgcagcgtag gtatggggga ggactggtca
      961 gagtgccact ctcccgcaac tctacacatg cgatgtactg ggtctctgga gcgaaaagca
     1021 acaccataaa aagtgtgtcc accacgagcc agctcctctt ggggcgcatg gacgggccta
     1081 ggaggccagt gaaatatgag gaggatgtga atctcggctc tggcacgcgg gctgtggtaa
     1141 gctgcgctga agctcccaac atgaagatca ttggtaaccg cattgaaagg atccgcagtg
     1201 agcacgcgga aacgtggttc tttgacgaga accacccata taggacatgg gcttaccatg
     1261 gaagctatga ggcccccaca caagggtcag cgtcctctct aataaacggg gttgtcaggc
     1321 tcctgtcaaa accctgggat gtggtgactg gagtcacagg aatagccatg accgacacca
     1381 caccgtatgg tcagcaaaga gttttcaagg aaaaagtgga cactagggtg ccagaccccc
     1441 aagaaggcac tcgtcaggtt atgagcatgg tctcttcctg gttgtggaaa gagctaggca
     1501 aacacaaacg gccacgagtc tgtaccaaag aagagttcat caacaaggtt cgtagcaatg
     1561 cagcattagg ggcaatattt gaagaggaaa aagagtggaa gactgcagtg gaagctgtga
     1621 acgatccaag gttctgggct ctagtggaca aggaaagaga gcaccacctg agaggagagt
     1681 gccagagttg tgtgtacaac atgatgggaa aaagagaaaa gaaacaaggg gaatttggaa
     1741 aggccaaggg cagccgcgcc atctggtata tgtggctagg ggctagattt ctagagttcg
     1801 aagcccttgg attcttgaac gaggatcact ggatggggag agagaactca ggaggtggtg
     1861 ttgaagggct gggattacaa agactcggat atgtcctaga agagatgagt cgcataccag
     1921 gaggaaggat gtatgcagat gacactgctg gctgggacac ccgcatcagc aggttcgatc
     1981 tggagaatga agctctaatc accaaccaaa tggagaaagg gcatagggcc ttggcattgg
     2041 ccataatcaa gtacacatac caaaacaaag tggtaaaggt ccttagacca gctgaaaaag
     2101 ggaaaacagt tatggacatt atttcgagac aagaccaaag ggggagcgga caagttgtca
     2161 cttacgctct taacacattt accaacctag tggtgcaact cattcggaat atggaggctg
     2221 aggaagttct agagatgcaa gacttgtggc tgctgcggag gtcagagaaa gtgaccaact
     2281 ggttgcagag caacggatgg gataggctca aacgaatggc agtcagtgga gatgattgcg
     2341 ttgtgaagcc aattgatgat aggtttgcac atgccctcag gttcttgaat gatatgggaa
     2401 aagttaggaa ggacacacaa gagtggaaac cctcaactgg atgggacaac tgggaagaag
     2461 ttccgttttg ctcccaccac ttcaacaagc tccatctcaa ggacgggagg tccattgtgg
     2521 ttccctgccg ccaccaagat gaactgattg gccgggcccg cgtctctcca ggggcgggat
     2581 ggagcatccg ggagactgct tgcctagcaa aatcatatgc gcaaatgtgg cagctccttt
     2641 atttccacag aagggacctc cgactgatgg ccaatgccat ttgttcatct gtgccagttg
     2701 actgggttcc aactgggaga actacctggt caatccatgg aaagggagaa tggatgacca
     2761 ctgaagacat gcttgtggtg tggaacagag tgtggattga ggagaacgac cacatggaag
     2821 acaagacccc agttacgaaa tggacagaca ttccctattt gggaaaaagg gaagacttgt
     2881 ggtgtggatc tctcataggg cacagaccgc gcaccacctg ggctgagaac attaaaaaca
     2941 cagtcaacat ggtgcgcagg atcataggtg atgaagaaaa gtacatggac tacctatcca
     3001 cccaagttcg ctacttgggt gaagaagggt ctacacctgg agtgctggac ccagctttct
     3061 tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg taagcctatc cctaaccctc
     3121 tcctcggtct cgattctacg cgtaccggtt agtaatgagt ttaaacgggg gaggctaact
     3181 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     3241 aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     3301 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     3361 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     3421 ggcaggccct gccatagcag atctgcgcag ctggggctct agggggtatc cccacgcgcc
     3481 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3541 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3601 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3661 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3721 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3781 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3841 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3901 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3961 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4021 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4081 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4141 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     4201 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     4261 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     4321 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4381 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4441 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     4501 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4561 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4621 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     4681 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     4741 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     4801 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     4861 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     4921 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     4981 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5041 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5101 ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc
     5161 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     5221 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     5281 caacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5341 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5401 ttatcatgtc tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct
     5461 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     5521 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     5581 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     5641 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     5701 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     5761 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     5821 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5881 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5941 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     6001 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     6061 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     6121 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     6181 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     6241 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     6301 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     6361 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     6421 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     6481 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     6541 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     6601 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     6661 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     6721 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     6781 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6841 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6901 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6961 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7021 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7081 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7141 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     7201 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     7261 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     7321 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     7381 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     7441 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     7501 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     7561 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtcg acggatcggg
     7621 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta
     7681 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt
     7741 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg
     7801 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact
     7861 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
     7921 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
     7981 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
     8041 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     8101 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     8161 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     8221 atg
//
