LOCUS Exported 8223 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Ectopic expression of ZIKV protein. ACCESSION . VERSION . KEYWORDS pcDNA3.2/NS5-Af-mut SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8223) 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 8223) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8223 /organism="synthetic DNA construct" /mol_type="other DNA" 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" CDS complement(1527..1538) /codon_start=1 /product="WELQut protease recognition and cleavage site" /label=WELQut site /translation="WELQ" protein_bind complement(3429..3453) /gene="mutant version of attB" /label=attB2 /bound_moiety="BP Clonase(TM)" /note="recombination site for the Gateway(R) BP reaction" CDS 3479..3520 /codon_start=1 /product="epitope tag from simian virus 5" /label=V5 tag /translation="GKPIPNPLLGLDST" primer_bind complement(3560..3579) /label=TK-pA-R /note="Thymidine kinase polyA, reverse primer" polyA_signal 3604..3652 /label=HSV TK poly(A) signal /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" rep_origin 3854..4282 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3941..3960) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4151..4172 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4291..4311) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4296..4625 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4476..4611 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4538..4557 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 4692..5486 /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(4746..4765) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5356..5375 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 5662..5783 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5699..5718) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5753..5772 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5832..5848) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(5832..5848) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(5845..5867) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 5856..5872 /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(5880..5910) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 5925..5946 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6063..6080) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6234..6822) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6314..6333) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6993..7853) /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 7616..7635 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7854..7958) /gene="bla" /label=AmpR promoter primer_bind complement(8033..8052) /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 ttcaggagga cagctatggg 721 gggtggaaca ggagagaccc tgggagagaa atggaaggcc cgcttgaacc agatgtcggc 781 cctggagttc tactcctaca aaaagtcagg catcaccgag gtgtgcagag aagaggcccg 841 ccgcgccctc aaggatggtg tggcaacggg aggccatgct gtgtcccgag gaagtgcaaa 901 gctgagatgg ttggtggagc ggggatacct gcagccctat ggaaaggtca ttgatcttgg 961 atgtggcaga gggggctgga gttactacgc cgccaccatc cgcaaagttc aagaagtgaa 1021 aggatacaca aaaggaggcc ctggtcatga agaacccatg ttggtgcaaa gctatgggtg 1081 gaacatagtc cgtcttaaga gtggggtgga cgtctttcat atggcggctg agccgtgtga 1141 cacgttgctg tgtgacatag gtgagtcatc atctagtcct gaagtggaag aagcacggac 1201 gctcagagtc ctctccatgg tgggggattg gcttgaaaaa agaccaggag ccttttgtat 1261 aaaagtgttg tgcccataca ccagcactat gatggaaacc ctggagcgac tgcagcgtag 1321 gtatggggga ggactggtca gagtgccact ctcccgcaac tctacacatg agatgtactg 1381 ggtctctgga gcgaaaagca acaccataaa aagtgtgtcc accacgagcc agctcctctt 1441 ggggcgcatg gacgggccta ggaggccagt gaaatatgag gaggatgtga atctcggctc 1501 tggcacgcgg gctgtggtaa gctgcgctga agctcccaac atgaagatca ttggtaaccg 1561 cattgaaagg atccgcagtg agcacgcgga aacgtggttc tttgacgaga accacccata 1621 taggacatgg gcttaccatg gaagctatga ggcccccaca caagggtcag cgtcctctct 1681 aataaacggg gttgtcaggc tcctgtcaaa accctgggat gtggtgactg gagtcacagg 1741 aatagccatg accgacacca caccgtatgg tcagcaaaga gttttcaagg aaaaagtgga 1801 cactagggtg ccagaccccc aagaaggcac tcgtcaggtt atgagcatgg tctcttcctg 1861 gttgtggaaa gagctaggca aacacaaacg gccacgagtc tgtaccaaag aagagttcat 1921 caacaaggtt cgtagcaatg cagcattagg ggcaatattt gaagaggaaa aagagtggaa 1981 gactgcagtg gaagctgtga acgatccaag gttctgggct ctagtggaca aggaaagaga 2041 gcaccacctg agaggagagt gccagagttg tgtgtacaac atgatgggaa aaagagaaaa 2101 gaaacaaggg gaatttggaa aggccaaggg cagccgcgcc atctggtata tgtggctagg 2161 ggctagattt ctagagttcg aagcccttgg attcttgaac gaggatcact ggatggggag 2221 agagaactca ggaggtggtg ttgaagggct gggattacaa agactcggat atgtcctaga 2281 agagatgagt cgcataccag gaggaaggat gtatgcagat gacactgctg gctgggacac 2341 ccgcatcagc aggttcgatc tggagaatga agctctaatc accaaccaaa tggagaaagg 2401 gcatagggcc ttggcattgg ccataatcaa gtacacatac caaaacaaag tggtaaaggt 2461 ccttagacca gctgaaaaag ggaaaacagt tatggacatt atttcgagac aagaccaaag 2521 ggggagcgga caagttgtca cttacgctct taacacattt accaacctag tggtgcaact 2581 cattcggaat atggaggctg aggaagttct agagatgcaa gacttgtggc tgctgcggag 2641 gtcagagaaa gtgaccaact ggttgcagag caacggatgg gataggctca aacgaatggc 2701 agtcagtgga gatgattgcg ttgtgaagcc aattgatgat aggtttgcac atgccctcag 2761 gttcttgaat gatatgggaa aagttaggaa ggacacacaa gagtggaaac cctcaactgg 2821 atgggacaac tgggaagaag ttccgttttg ctcccaccac ttcaacaagc tccatctcaa 2881 ggacgggagg tccattgtgg ttccctgccg ccaccaagat gaactgattg gccgggcccg 2941 cgtctctcca ggggcgggat ggagcatccg ggagactgct tgcctagcaa aatcatatgc 3001 gcaaatgtgg cagctccttt atttccacag aagggacctc cgactgatgg ccaatgccat 3061 ttgttcatct gtgccagttg actgggttcc aactgggaga actacctggt caatccatgg 3121 aaagggagaa tggatgacca ctgaagacat gcttgtggtg tggaacagag tgtggattga 3181 ggagaacgac cacatggaag acaagacccc agttacgaaa tggacagaca ttccctattt 3241 gggaaaaagg gaagacttgt ggtgtggatc tctcataggg cacagaccgc gcaccacctg 3301 ggctgagaac attaaaaaca cagtcaacat gatgcgcagg atcataggtg atgaagaaaa 3361 gtacatggac tacctatcca cccaagttcg ctacttgggt gaagaagggt ctacacctgg 3421 agtgctggac ccagctttct tgtacaaagt ggttgatcta gagggcccgc ggttcgaagg 3481 taagcctatc cctaaccctc tcctcggtct cgattctacg cgtaccggtt agtaatgagt 3541 ttaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta 3601 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc 3661 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa 3721 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg 3781 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct 3841 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg 3901 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct 3961 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta 4021 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt 4081 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg 4141 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat 4201 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt 4261 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt 4321 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca 4381 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt 4441 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt 4501 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg 4561 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt 4621 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag 4681 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg 4741 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt 4801 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc 4861 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt 4921 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag 4981 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg 5041 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag 5101 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg 5161 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc 5221 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca 5281 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc 5341 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg 5401 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct 5461 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa 5521 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg 5581 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg 5641 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc 5701 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg 5761 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg 5821 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac 5881 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc 5941 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg 6001 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct 6061 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac 6121 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga 6181 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat 6241 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac 6301 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct 6361 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg 6421 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg 6481 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt 6541 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg 6601 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac 6661 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga 6721 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt 6781 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt 6841 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga 6901 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc 6961 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct 7021 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata 7081 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca 7141 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga 7201 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga 7261 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg 7321 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga 7381 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt 7441 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct 7501 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca 7561 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat 7621 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga 7681 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc 7741 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg 7801 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc 7861 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt 7921 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca 7981 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct 8041 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg 8101 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga 8161 agaatctgct tagggttagg cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc 8221 gtt //