LOCUS       sequence-234569-        5988 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 5988)
  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 5988)
  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
FEATURES             Location/Qualifiers
     source          1..5988
                     /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(1194..1218)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1244..1285
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1325..1344)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1369..1417
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1619..2047
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1706..1725)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1916..1937
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2056..2076)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2061..2390
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2241..2376
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2303..2322
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2457..3251
                     /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(2511..2530)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3121..3140
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3427..3548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3464..3483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3518..3537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3597..3613)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3597..3613)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3610..3632)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3621..3637
                     /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(3645..3651,3652..3669,3670..3675))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3690..3711
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3828..3845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3999..4587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4079..4098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4758..5549,5550..5618))
                     /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     5381..5400
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5619..5723)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5798..5817)
                     /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 aagggcgaat tcgacccagc
     1201 tttcttgtac aaagtggttg atctagaggg cccgcggttc gaaggtaagc ctatccctaa
     1261 ccctctcctc ggtctcgatt ctacgcgtac cggttagtaa tgagtttaaa cgggggaggc
     1321 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     1381 gacagaataa aacgcacggg tgttgggtcg tttgttcata aacgcggggt tcggtcccag
     1441 ggctggcact ctgtcgatac cccaccgaga ccccattggg gccaatacgc ccgcgtttct
     1501 tccttttccc caccccaccc cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc
     1561 ggggcggcag gccctgccat agcagatctg cgcagctggg gctctagggg gtatccccac
     1621 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1681 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1741 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1801 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1861 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1921 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1981 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     2041 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     2101 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     2161 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2221 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2281 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2341 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2401 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2461 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2521 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2581 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     2641 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2701 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2761 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2821 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2881 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2941 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     3001 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     3061 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     3121 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3181 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3241 agttcttctg agcgggactc tggggttcgc gaaatgaccg accaagcgac gcccaacctg
     3301 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3361 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3421 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3481 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3541 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     3601 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3661 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3721 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3781 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3841 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3901 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3961 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4021 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4081 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4141 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4201 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4261 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4321 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4381 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4441 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4501 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4561 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4621 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4681 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4741 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4801 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4861 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4921 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4981 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     5041 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5101 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5161 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5221 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5281 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5341 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5401 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5461 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5521 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5581 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5641 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5701 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     5761 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     5821 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     5881 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     5941 ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
