LOCUS       sequence-234610-        6741 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 6741)
  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 6741)
  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/NS4A-2K-NS4B
FEATURES             Location/Qualifiers
     source          1..6741
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    677..701
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(1947..1971)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1997..2038
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(2078..2097)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    2122..2170
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2372..2800
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2459..2478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2669..2690
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2809..2829)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2814..3143
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2994..3129
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3056..3075
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3210..4004
                     /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(3264..3283)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3874..3893
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4180..4301
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4217..4236)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4271..4290
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4350..4366)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4350..4366)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4363..4385)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4374..4390
                     /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(4398..4404,4405..4422,4423..4428))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4443..4464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4581..4598)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4752..5340)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4832..4851)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5511..6302,6303..6371))
                     /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     6134..6153
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6372..6476)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6551..6570)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag ctatcaacaa gtttgtacaa aaaagcaggc tccgaattcg cccttaggag
      721 gacagctatg ggagcggctt ttggagtgat ggaagccctg ggaacactgc caggacacat
      781 gacagagaga ttccaggaag ccattgacaa cctcgctgtg ctcatgcggg cagagactgg
      841 aagcaggcct tacaaagccg cggcggccca attgccggag accctagaga ccattatgct
      901 tttggggttg ctgggaacag tctcgctggg aatctttttc gtcttgatga ggaacaaggg
      961 catagggaag atgggctttg gaatggtgac tcttggggcc agcgcatggc tcatgtggct
     1021 ctcggaaatt gagccagcca gaattgcatg tgtcctcatt gttgtgttcc tattgctggt
     1081 ggtgctcata cctgagccag aaaagcaaag atctccccag gacaaccaaa tggcaatcat
     1141 catcatggta gcagtaggtc ttctgggctt gattaccgcc aatgaactcg gatggttgga
     1201 gagaacaaag agtgacctaa gccatctaat gggaaggaga gaggagggag caaccatagg
     1261 attctcaatg gacattgacc tgcggccagc ctcagcttgg gccatctatg ctgccttgac
     1321 aactttcatt accccagccg tccaacatgc agtgaccact tcatacaaca actactcctt
     1381 aatggcgatg gccacgcaag ctggagtgtt gtttggtatg ggcaaaggga tgccattcta
     1441 cgcatgggac tttggagtcc cgctgctaat gataggttgc tactcacaat taacacccct
     1501 gaccctaata gtggccatca ttttgctcgt ggcgcactac atgtacttga tcccagggct
     1561 gcaggcagca gctgcgcgtg ctgcccagaa gagaacggca gctggcatca tgaagaaccc
     1621 tgttgtggat ggaatagtgg tgactgacat tgacacaatg acaattgacc cccaagtgga
     1681 gaaaaagatg ggacaggtgc tactcatagc agtagcagtc tccagcgcca tactgtcgcg
     1741 gaccgcctgg gggtgggggg aggctggggc cctgatcaca gccgcaactt ccactttgtg
     1801 ggaaggctct ccgaacaagt actggaactc ctctacagcc acttcactgt gtaacatttt
     1861 taggggaagt tacttggctg gagcttctct aatctacata gtaacaagaa acgctggctt
     1921 ggtcaagaga cgtaagggcg aattcgaccc agctttcttg tacaaagtgg ttgatctaga
     1981 gggcccgcgg ttcgaaggta agcctatccc taaccctctc ctcggtctcg attctacgcg
     2041 taccggttag taatgagttt aaacggggga ggctaactga aacacggaag gagacaatac
     2101 cggaaggaac ccgcgctatg acggcaataa aaagacagaa taaaacgcac gggtgttggg
     2161 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     2221 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     2281 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcagat
     2341 ctgcgcagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
     2401 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     2461 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     2521 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     2581 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     2641 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     2701 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     2761 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     2821 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2881 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2941 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     3001 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     3061 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     3121 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     3181 atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt
     3241 ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct
     3301 gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga
     3361 ccgacctgtc cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg
     3421 ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact
     3481 ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg
     3541 agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct
     3601 gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg
     3661 gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt
     3721 tcgccaggct caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg
     3781 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     3841 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     3901 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     3961 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt
     4021 cgcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4081 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4141 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     4201 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     4261 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     4321 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     4381 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     4441 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4501 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4561 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     4621 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     4681 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4741 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4801 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4861 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4921 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4981 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5041 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5101 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     5161 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     5221 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     5281 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     5341 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     5401 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     5461 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     5521 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     5581 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     5641 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     5701 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5761 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5821 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5881 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5941 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6001 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6061 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6121 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     6181 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     6241 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     6301 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     6361 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     6421 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     6481 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     6541 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     6601 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     6661 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     6721 tacgggccag atatacgcgt t
//
