LOCUS       sequence-234616-        6360 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 6360)
  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 6360)
  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: pcDNA3.2/2K-NS4B
FEATURES             Location/Qualifiers
     source          1..6360
                     /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(1566..1590)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1616..1657
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(1697..1716)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1741..1789
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1991..2419
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2078..2097)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2288..2309
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2428..2448)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2433..2762
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2613..2748
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2675..2694
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2829..3623
                     /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(2883..2902)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3493..3512
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3799..3920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3836..3855)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3890..3909
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3969..3985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3969..3985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3982..4004)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3993..4009
                     /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(4017..4023,4024..4041,4042..4047))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4062..4083
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4200..4217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4371..4959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4451..4470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5130..5921,5922..5990))
                     /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     5753..5772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5991..6095)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6170..6189)
                     /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 tctccccagg acaaccaaat ggcaatcatc atcatggtag cagtaggtct
      781 tctgggcttg attaccgcca atgaactcgg atggttggag agaacaaaga gtgacctaag
      841 ccatctaatg ggaaggagag aggagggagc aaccatagga ttctcaatgg acattgacct
      901 gcggccagcc tcagcttggg ccatctatgc tgccttgaca actttcatta ccccagccgt
      961 ccaacatgca gtgaccactt catacaacaa ctactcctta atggcgatgg ccacgcaagc
     1021 tggagtgttg tttggtatgg gcaaagggat gccattctac gcatgggact ttggagtccc
     1081 gctgctaatg ataggttgct actcacaatt aacacccctg accctaatag tggccatcat
     1141 tttgctcgtg gcgcactaca tgtacttgat cccagggctg caggcagcag ctgcgcgtgc
     1201 tgcccagaag agaacggcag ctggcatcat gaagaaccct gttgtggatg gaatagtggt
     1261 gactgacatt gacacaatga caattgaccc ccaagtggag aaaaagatgg gacaggtgct
     1321 actcatagca gtagcagtct ccagcgccat actgtcgcgg accgcctggg ggtgggggga
     1381 ggctggggcc ctgatcacag ccgcaacttc cactttgtgg gaaggctctc cgaacaagta
     1441 ctggaactcc tctacagcca cttcactgtg taacattttt aggggaagtt acttggctgg
     1501 agcttctcta atctacatag taacaagaaa cgctggcttg gtcaagagac gtaagggcga
     1561 attcgaccca gctttcttgt acaaagtggt tgatctagag ggcccgcggt tcgaaggtaa
     1621 gcctatccct aaccctctcc tcggtctcga ttctacgcgt accggttagt aatgagttta
     1681 aacgggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     1741 cggcaataaa aagacagaat aaaacgcacg ggtgttgggt cgtttgttca taaacgcggg
     1801 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     1861 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     1921 gcagccaacg tcggggcggc aggccctgcc atagcagatc tgcgcagctg gggctctagg
     1981 gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc
     2041 agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc
     2101 tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg
     2161 ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca
     2221 cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc
     2281 tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     2341 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     2401 caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     2461 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     2521 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2581 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2641 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2701 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2761 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     2821 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     2881 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     2941 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     3001 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     3061 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     3121 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     3181 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     3241 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     3301 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     3361 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     3421 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     3481 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     3541 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     3601 gccttcttga cgagttcttc tgagcgggac tctggggttc gcgaaatgac cgaccaagcg
     3661 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3721 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3781 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3841 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3901 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     3961 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     4021 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     4081 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     4141 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4561 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4801 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6121 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
     6181 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
     6241 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
     6301 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
//
