LOCUS       sequence-241007-        6291 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 6291)
  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 6291)
  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/NS4B
FEATURES             Location/Qualifiers
     source          1..6291
                     /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"
     protein_bind    complement(1117..1141)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1167..1208
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1248..1267)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1292..1340
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1542..1970
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1629..1648)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1839..1860
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1979..1999)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1984..2313
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2164..2299
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2226..2245
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2380..3174
                     /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(2434..2453)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3044..3063
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3350..3471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3387..3406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3441..3460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3520..3536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3520..3536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3533..3555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3544..3560
                     /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(3568..3574,3575..3592,3593..3598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3613..3634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3751..3768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3922..4510)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4002..4021)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4681..5472,5473..5541))
                     /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     5304..5323
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5542..5646)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5721..5740)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        5912..6291
                     /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 tccgaattcg cccttaggag gacagctatg aatgaactcg
      361 gatggttgga gagaacaaag agtgacctaa gccatctaat gggaaggaga gaggagggag
      421 caaccatagg attctcaatg gacattgacc tgcggccagc ctcagcttgg gccatctatg
      481 ctgccttgac aactttcatt accccagccg tccaacatgc agtgaccact tcatacaaca
      541 actactcctt aatggcgatg gccacgcaag ctggagtgtt gtttggtatg ggcaaaggga
      601 tgccattcta cgcatgggac tttggagtcc cgctgctaat gataggttgc tactcacaat
      661 taacacccct gaccctaata gtggccatca ttttgctcgt ggcgcactac atgtacttga
      721 tcccagggct gcaggcagca gctgcgcgtg ctgcccagaa gagaacggca gctggcatca
      781 tgaagaaccc tgttgtggat ggaatagtgg tgactgacat tgacacaatg acaattgacc
      841 cccaagtgga gaaaaagatg ggacaggtgc tactcatagc agtagcagtc tccagcgcca
      901 tactgtcgcg gaccgcctgg gggtgggggg aggctggggc cctgatcaca gccgcaactt
      961 ccactttgtg ggaaggctct ccgaacaagt actggaactc ctctacagcc acttcactgt
     1021 gtaacatttt taggggaagt tacttggctg gagcttctct aatctacata gtaacaagaa
     1081 acgctggctt ggtcaagaga cgtaagggcg aattcgaccc agctttcttg tacaaagtgg
     1141 ttgatctaga gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg
     1201 attctacgcg taccggttag taatgagttt aaacggggga ggctaactga aacacggaag
     1261 gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac
     1321 gggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     1381 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     1441 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     1501 catagcagat ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc
     1561 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1621 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1681 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1741 ccccaaaaaa cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1801 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1861 aacaacactc aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc
     1921 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg
     1981 aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2041 agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc
     2101 agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2161 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2221 ttttttattt atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga
     2281 ggaggctttt ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt
     2341 ttcggatctg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     2401 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     2461 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     2521 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta
     2581 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     2641 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     2701 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     2761 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     2821 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     2881 gccgaactgt tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc
     2941 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     3001 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     3061 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     3121 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     3181 ctctggggtt cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     3241 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     3301 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     3361 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     3421 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     3481 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     3541 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     3601 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     3661 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     3721 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     3781 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     3841 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     3901 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     3961 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     4021 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     4081 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     4141 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     4201 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     4261 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     4321 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     4381 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     4441 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     4501 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     4561 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     4621 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     4681 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     4741 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     4801 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     4861 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     4921 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     4981 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     5041 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     5101 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     5161 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     5221 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     5281 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     5341 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     5401 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     5461 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     5521 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     5581 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     5641 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     5701 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     5761 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     5821 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     5881 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag
     5941 taatcaatta cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt
     6001 acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg
     6061 acgtatgttc ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat
     6121 ttacggtaaa ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct
     6181 attgacgtca atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg
     6241 gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat g
//
