LOCUS       sequence-365329-        7224 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  The RdRp domain (with a 6-histidine tag) from Zika virus 
            nonstructural protein 5 (French Polynesia strain, GenBank accession 
            number ARU07075.1) encompassing residues 270 to 89.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7224)
  AUTHORS   Gharbi-Ayachi A, Santhanakrishnan S, Wong YH, Chan KWK, Tan ST, 
            Bates RW, Vasudevan SG, El Sahili A, Lescar J
  TITLE     Non-nucleoside Inhibitors of Zika Virus RNA-Dependent RNA 
            Polymerase.
  JOURNAL   J Virol. 2020 Oct 14;94(21). pii: JVI.00794-20. doi: 
            10.1128/JVI.00794-20. Print 2020 Oct 14.
  PUBMED    32796069
REFERENCE   2  (bases 1 to 7224)
  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: "J Virol."; 
            date: "2020-10-14"; volume: "94(21). pii: JVI.00794-20. doi"; pages:
            " 10.1128/JVI.00794-20. Print 2020 Oct 14"
COMMENT     Sequence Label: pNIC28-Bsa4_ZV-NS5-RdRp
FEATURES             Location/Qualifiers
     source          1..7224
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             134..154
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             2089..2106
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2159..2177)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2173..2220
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2257..2712
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2344..2363)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2554..2575
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2804..3619)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(2804..3619)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     3527..3546
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3611..3630
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3741..4329
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4230..4249
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4483..4500
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4515..4654
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4580..4599)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4740..4762
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4756..4947)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5756..6838)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     6800..6819
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6839..6916)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7122..7141
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atatacatat gcaccatcat catcatcatt cttctggtgt
      121 agatctgggt accgagaacc tgtacttcca atccatggcg gaggcgccga acatgaaaat
      181 cattggtaac cgtatcgaac gtattcgtag cgaacacgcg gaaacctggt tctttgacga
      241 gaaccacccg taccgtacct gggcgtacca cggtagctat gaagcgccga cccagggcag
      301 cgcgagcagc ctgatcaacg gtgtggttcg tctgctgagc aagccgtggg acgtggttac
      361 cggtgtgacc ggcattgcga tgaccgatac caccccgtac ggtcagcaac gtgttttcaa
      421 ggagaaagtg gacacccgtg ttccggaccc gcaggaaggt acccgtcaag tgatgagcat
      481 ggttagcagc tggctgtgga aagaactggg caagcacaaa cgtccgcgtg tgtgcaccaa
      541 agaggagttc atcaacaaag ttcgtagcaa cgcggcgctg ggtgcgattt ttgaggaaga
      601 gaaggaatgg aaaaccgcgg tggaggcggt taacgacccg cgtttttggg cgctggttga
      661 caaggaacgt gagcaccacc tgcgtggcga gtgccagagc tgcgtttaca acatgatggg
      721 taaacgtgaa aagaaacaag gcgagttcgg caaggcgaaa ggcagccgtg cgatctggta
      781 tatgtggctg ggtgcgcgtt tcctggaatt tgaggcgctg ggctttctga acgaagacca
      841 ctggatgggt cgtgagaaca gcggcggtgg cgtggaaggt ctgggtctgc agcgtctggg
      901 ctacgttctg gaagagatga gccgtatccc gggtggccgt atgtatgcgg acgataccgc
      961 gggttgggac acccgtatca gccgtttcga tctggaaaac gaggcgctga ttaccaacca
     1021 gatggaaaaa ggtcaccgtg cgctggcgct ggcgatcatt aaatacacct atcaaaacaa
     1081 ggtggttaaa gtgctgcgtc cggcggagaa gggtaaaacc gttatggaca tcattagccg
     1141 tcaggatcaa cgtggtagcg gccaggtggt tacctacgcg ctgaacacct ttaccaacct
     1201 ggtggttcag ctgatccgta acatggaagc ggaagaggtg ctggagatgc aagacctgtg
     1261 gctgctgcgt cgtagcgaaa aggttaccaa ctggctgcaa agcaacggct gggatcgtct
     1321 gaagcgtatg gcggttagcg gtgacgattg cgtggttaaa ccgattgacg atcgtttcgc
     1381 gcacgcgctg cgttttctga acgacatggg caaagtgcgt aaagataccc aggagtggaa
     1441 accgagcacc ggttgggaca actgggaaga ggttccgttc tgcagccacc actttaacaa
     1501 gctgcacctg aaagacggcc gtagcatcgt ggttccgtgc cgtcaccaag atgagctgat
     1561 tggtcgtgcg cgtgtgagcc cgggtgcggg ttggagcatc cgtgaaaccg cgtgcctggc
     1621 gaaaagctac gcgcagatgt ggcaactgct gtatttccac cgtcgtgacc tgcgtctgat
     1681 ggcgaacgcg atctgcagca gcgtgccggt tgattgggtt ccgaccggtc gtaccacctg
     1741 gagcattcac ggcaagggcg aatggatgac caccgaggac atgctggttg tgtggaaccg
     1801 tgtgtggatc gaagagaacg accacatgga ggataagacc ccggttacca aatggaccga
     1861 cattccgtac ctgggcaaac gtgaagatct gtggtgcggt agcctgattg gtcaccgtcc
     1921 gcgtaccacc tgggcggaga acattaagaa caccgtgaac atggttcgtc gtatcattgg
     1981 tgatgaagag aaatatatgg actacctgag cacccaagtt cgttgacagt aaaggtggat
     2041 acggatccga attcgagctc cgtcgacaag cttgcggccg cactcgagca ccaccaccac
     2101 caccactgag atccggctgc taacaaagcc cgaaaggaag ctgagttggc tgctgccacc
     2161 gctgagcaat aactagcata accccttggg gcctctaaac gggtcttgag gggttttttg
     2221 ctgaaaggag gaactatatc cggattggcg aatgggacgc gccctgtagc ggcgcattaa
     2281 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     2341 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     2401 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     2461 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     2521 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     2581 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     2641 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa
     2701 cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2761 tttctaaata cattcaaata tgtatccgct catgaattaa ttcttagaaa aactcatcga
     2821 gcatcaaatg aaactgcaat ttattcatat caggattatc aataccatat ttttgaaaaa
     2881 gccgtttctg taatgaagga gaaaactcac cgaggcagtt ccataggatg gcaagatcct
     2941 ggtatcggtc tgcgattccg actcgtccaa catcaataca acctattaat ttcccctcgt
     3001 caaaaataag gttatcaagt gagaaatcac catgagtgac gactgaatcc ggtgagaatg
     3061 gcaaaagttt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
     3121 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcgagacgaa
     3181 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
     3241 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
     3301 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
     3361 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
     3421 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
     3481 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
     3541 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctagag caagacgttt
     3601 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
     3661 ttgttcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3721 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3781 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     3841 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     3901 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     3961 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4021 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4081 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4141 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4201 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4261 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4321 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4381 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4441 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4501 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4561 cgcaatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatacac
     4621 tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa cacccgctga
     4681 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     4741 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga ggcagctgcg
     4801 gtaaagctca tcagcgtggt cgtgaagcga ttcacagatg tctgcctgtt catccgcgtc
     4861 cagctcgttg agtttctcca gaagcgttaa tgtctggctt ctgataaagc gggccatgtt
     4921 aagggcggtt ttttcctgtt tggtcactga tgcctccgtg taagggggat ttctgttcat
     4981 gggggtaatg ataccgatga aacgagagag gatgctcacg atacgggtta ctgatgatga
     5041 acatgcccgg ttactggaac gttgtgaggg taaacaactg gcggtatgga tgcggcggga
     5101 ccagagaaaa atcactcagg gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc
     5161 acagggtagc cagcagcatc ctgcgatgca gatccggaac ataatggtgc agggcgctga
     5221 cttccgcgtt tccagacttt acgaaacacg gaaaccgaag accattcatg ttgttgctca
     5281 ggtcgcagac gttttgcagc agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt
     5341 ctgctaacca gtaaggcaac cccgccagcc tagccgggtc ctcaacgaca ggagcacgat
     5401 catgcgcacc cgtggggccg ccatgccggc gataatggcc tgcttctcgc cgaaacgttt
     5461 ggtggcggga ccagtgacga aggcttgagc gagggcgtgc aagattccga ataccgcaag
     5521 cgacaggccg atcatcgtcg cgctccagcg aaagcggtcc tcgccgaaaa tgacccagag
     5581 cgctgccggc acctgtccta cgagttgcat gataaagaag acagtcataa gtgcggcgac
     5641 gatagtcatg ccccgcgccc accggaagga gctgactggg ttgaaggctc tcaagggcat
     5701 cggtcgagat cccggtgcct aatgagtgag ctaacttaca ttaattgcgt tgcgctcact
     5761 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     5821 ggggagaggc ggtttgcgta ttgggcgcca gggtggtttt tcttttcacc agtgagacgg
     5881 gcaacagctg attgcccttc accgcctggc cctgagagag ttgcagcaag cggtccacgc
     5941 tggtttgccc cagcaggcga aaatcctgtt tgatggtggt taacggcggg atataacatg
     6001 agctgtcttc ggtatcgtcg tatcccacta ccgagatatc cgcaccaacg cgcagcccgg
     6061 actcggtaat ggcgcgcatt gcgcccagcg ccatctgatc gttggcaacc agcatcgcag
     6121 tgggaacgat gccctcattc agcatttgca tggtttgttg aaaaccggac atggcactcc
     6181 agtcgccttc ccgttccgct atcggctgaa tttgattgcg agtgagatat ttatgccagc
     6241 cagccagacg cagacgcgcc gagacagaac ttaatgggcc cgctaacagc gcgatttgct
     6301 ggtgacccaa tgcgaccaga tgctccacgc ccagtcgcgt accgtcttca tgggagaaaa
     6361 taatactgtt gatgggtgtc tggtcagaga catcaagaaa taacgccgga acattagtgc
     6421 aggcagcttc cacagcaatg gcatcctggt catccagcgg atagttaatg atcagcccac
     6481 tgacgcgttg cgcgagaaga ttgtgcaccg ccgctttaca ggcttcgacg ccgcttcgtt
     6541 ctaccatcga caccaccacg ctggcaccca gttgatcggc gcgagattta atcgccgcga
     6601 caatttgcga cggcgcgtgc agggccagac tggaggtggc aacgccaatc agcaacgact
     6661 gtttgcccgc cagttgttgt gccacgcggt tgggaatgta attcagctcc gccatcgccg
     6721 cttccacttt ttcccgcgtt ttcgcagaaa cgtggctggc ctggttcacc acgcgggaaa
     6781 cggtctgata agagacaccg gcatactctg cgacatcgta taacgttact ggtttcacat
     6841 tcaccaccct gaattgactc tcttccgggc gctatcatgc cataccgcga aaggttttgc
     6901 gccattcgat ggtgtccggg atctcgacgc tctcccttat gcgactcctg cattaggaag
     6961 cagcccagta gtaggttgag gccgttgagc accgccgccg caaggaatgg tgcatgcaag
     7021 gagatggcgc ccaacagtcc cccggccacg gggcctgcca ccatacccac gccgaaacaa
     7081 gcgctcatga gcccgaagtg gcgagcccga tcttccccat cggtgatgtc ggcgatatag
     7141 gcgccagcaa ccgcacctgt ggcgccggtg atgccggcca cgatgcgtcc ggcgtagagg
     7201 atcgagatct cgatcccgcg aaat
//
