LOCUS       sequence-229600-        8221 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 8221)
  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 8221)
  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-Af-mut
FEATURES             Location/Qualifiers
     source          1..8221
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    911..935
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(1759..1770)
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     protein_bind    complement(3661..3685)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3711..3752
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(3792..3811)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3836..3884
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4086..4514
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4173..4192)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4383..4404
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4523..4543)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4528..4857
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4708..4843
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4770..4789
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4924..5718
                     /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(4978..4997)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5588..5607
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5894..6015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5931..5950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5985..6004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6064..6080)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6064..6080)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6077..6099)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6088..6104
                     /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(6112..6118,6119..6136,6137..6142))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6157..6178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6295..6312)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6466..7054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6546..6565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7225..8016,8017..8085))
                     /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     7848..7867
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8086..8190)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagctatca acaagtttgt acaaaaaagc aggctaggag gacagctatg gggggtggaa
      961 caggagagac cctgggagag aaatggaagg cccgcttgaa ccagatgtcg gccctggagt
     1021 tctactccta caaaaagtca ggcatcaccg aggtgtgcag agaagaggcc cgccgcgccc
     1081 tcaaggatgg tgtggcaacg ggaggccatg ctgtgtcccg aggaagtgca aagctgagat
     1141 ggttggtgga gcggggatac ctgcagccct atggaaaggt cattgatctt ggatgtggca
     1201 gagggggctg gagttactac gccgccacca tccgcaaagt tcaagaagtg aaaggataca
     1261 caaaaggagg ccctggtcat gaagaaccca tgttggtgca aagctatggg tggaacatag
     1321 tccgtcttaa gagtggggtg gacgtctttc atatggcggc tgagccgtgt gacacgttgc
     1381 tgtgtgacat aggtgagtca tcatctagtc ctgaagtgga agaagcacgg acgctcagag
     1441 tcctctccat ggtgggggat tggcttgaaa aaagaccagg agccttttgt ataaaagtgt
     1501 tgtgcccata caccagcact atgatggaaa ccctggagcg actgcagcgt aggtatgggg
     1561 gaggactggt cagagtgcca ctctcccgca actctacaca tgagatgtac tgggtctctg
     1621 gagcgaaaag caacaccata aaaagtgtgt ccaccacgag ccagctcctc ttggggcgca
     1681 tggacgggcc taggaggcca gtgaaatatg aggaggatgt gaatctcggc tctggcacgc
     1741 gggctgtggt aagctgcgct gaagctccca acatgaagat cattggtaac cgcattgaaa
     1801 ggatccgcag tgagcacgcg gaaacgtggt tctttgacga gaaccaccca tataggacat
     1861 gggcttacca tggaagctat gaggccccca cacaagggtc agcgtcctct ctaataaacg
     1921 gggttgtcag gctcctgtca aaaccctggg atgtggtgac tggagtcaca ggaatagcca
     1981 tgaccgacac cacaccgtat ggtcagcaaa gagttttcaa ggaaaaagtg gacactaggg
     2041 tgccagaccc ccaagaaggc actcgtcagg ttatgagcat ggtctcttcc tggttgtgga
     2101 aagagctagg caaacacaaa cggccacgag tctgtaccaa agaagagttc atcaacaagg
     2161 ttcgtagcaa tgcagcatta ggggcaatat ttgaagagga aaaagagtgg aagactgcag
     2221 tggaagctgt gaacgatcca aggttctggg ctctagtgga caaggaaaga gagcaccacc
     2281 tgagaggaga gtgccagagt tgtgtgtaca acatgatggg aaaaagagaa aagaaacaag
     2341 gggaatttgg aaaggccaag ggcagccgcg ccatctggta tatgtggcta ggggctagat
     2401 ttctagagtt cgaagccctt ggattcttga acgaggatca ctggatgggg agagagaact
     2461 caggaggtgg tgttgaaggg ctgggattac aaagactcgg atatgtccta gaagagatga
     2521 gtcgcatacc aggaggaagg atgtatgcag atgacactgc tggctgggac acccgcatca
     2581 gcaggttcga tctggagaat gaagctctaa tcaccaacca aatggagaaa gggcataggg
     2641 ccttggcatt ggccataatc aagtacacat accaaaacaa agtggtaaag gtccttagac
     2701 cagctgaaaa agggaaaaca gttatggaca ttatttcgag acaagaccaa agggggagcg
     2761 gacaagttgt cacttacgct cttaacacat ttaccaacct agtggtgcaa ctcattcgga
     2821 atatggaggc tgaggaagtt ctagagatgc aagacttgtg gctgctgcgg aggtcagaga
     2881 aagtgaccaa ctggttgcag agcaacggat gggataggct caaacgaatg gcagtcagtg
     2941 gagatgattg cgttgtgaag ccaattgatg ataggtttgc acatgccctc aggttcttga
     3001 atgatatggg aaaagttagg aaggacacac aagagtggaa accctcaact ggatgggaca
     3061 actgggaaga agttccgttt tgctcccacc acttcaacaa gctccatctc aaggacggga
     3121 ggtccattgt ggttccctgc cgccaccaag atgaactgat tggccgggcc cgcgtctctc
     3181 caggggcggg atggagcatc cgggagactg cttgcctagc aaaatcatat gcgcaaatgt
     3241 ggcagctcct ttatttccac agaagggacc tccgactgat ggccaatgcc atttgttcat
     3301 ctgtgccagt tgactgggtt ccaactggga gaactacctg gtcaatccat ggaaagggag
     3361 aatggatgac cactgaagac atgcttgtgg tgtggaacag agtgtggatt gaggagaacg
     3421 accacatgga agacaagacc ccagttacga aatggacaga cattccctat ttgggaaaaa
     3481 gggaagactt gtggtgtgga tctctcatag ggcacagacc gcgcaccacc tgggctgaga
     3541 acattaaaaa cacagtcaac atgatgcgca ggatcatagg tgatgaagaa aagtacatgg
     3601 actacctatc cacccaagtt cgctacttgg gtgaagaagg gtctacacct ggagtgctgg
     3661 acccagcttt cttgtacaaa gtggttgatc tagagggccc gcggttcgaa ggtaagccta
     3721 tccctaaccc tctcctcggt ctcgattcta cgcgtaccgg ttagtaatga gtttaaacgg
     3781 gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca
     3841 ataaaaagac agaataaaac gcacgggtgt tgggtcgttt gttcataaac gcggggttcg
     3901 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     3961 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4021 caacgtcggg gcggcaggcc ctgccatagc agatctgcgc agctggggct ctagggggta
     4081 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4141 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4201 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     4261 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4321 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4381 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4441 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4501 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4561 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4621 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4681 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4741 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4801 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4861 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4921 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4981 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5041 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5101 ctgcaggacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5161 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5221 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     5281 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     5341 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     5401 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc
     5461 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     5521 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     5581 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5641 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5701 cttgacgagt tcttctgagc gggactctgg ggttcgcgaa atgaccgacc aagcgacgcc
     5761 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     5821 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     5881 cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     5941 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6001 catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc
     6061 atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg
     6121 agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat
     6181 tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg
     6241 aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct
     6301 cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc
     6361 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     6421 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     6481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     6541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     6601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     6661 atgctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     6721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     6781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     6841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     6901 tagaaggaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     6961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     7021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     7081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     7141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     7201 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     7261 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     7321 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     7381 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     7441 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     7501 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     7561 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     7621 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     7681 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     7741 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     7801 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     7861 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     7921 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     7981 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8041 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8101 atattattga agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat
     8161 ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt
     8221 c
//
