LOCUS       sequence-175197-        8028 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of dengue virus 3 NS5 protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8028)
  AUTHORS   Zhao Y, Soh TS, Zheng J, Chan KW, Phoo WW, Lee CC, Tay MY, 
            Swaminathan K, Cornvik TC, Lim SP, Shi PY, Lescar J, Vasudevan SG, 
            Luo D
  TITLE     A crystal structure of the Dengue virus NS5 protein reveals a novel 
            inter-domain interface essential for protein flexibility and virus 
            replication.
  JOURNAL   PLoS Pathog. 2015 Mar 16;11(3):e1004682. doi: 
            10.1371/journal.ppat.1004682. eCollection 2015 Mar.
  PUBMED    25775415
REFERENCE   2  (bases 1 to 8028)
  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: "PLoS 
            Pathog."; date: "2015-03-16"; volume: "11(3):e1004682. doi"; pages: 
            " 10.1371/journal.ppat.1004682. eCollection 2015 Mar"
COMMENT     Sequence Label: pNIC28-DENV3-NS5FL
FEATURES             Location/Qualifiers
     source          1..8028
                     /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             2893..2910
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2963..2981)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2977..3024
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      3061..3516
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3148..3167)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3358..3379
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3608..4423)
                     /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(3608..4423)
                     /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     4331..4350
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4415..4434
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4545..5133
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5034..5053
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5287..5304
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5319..5458
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5384..5403)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5544..5566
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5560..5751)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6560..7642)
                     /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     7604..7623
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7643..7720)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7926..7945
                     /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 atccatgggt gaaactttag gagaaaaatg
      181 gaaaaagaaa ttaaatcaat tatcccggaa agagtttgac ctttacaaga aatctggaat
      241 cactgaagtg gatagaacag aagccaaaga agggctgaaa agaggagaga caacacatca
      301 tgctgtgtcc cgaggtagcg caaaactcca atggtttgtg gaaagaaaca tggtcgttcc
      361 cgaaggaaga gtcatagact tgggctgcgg aagaggaggc tggtcatatt actgtgcagg
      421 actgaaaaaa gtcacagaag tgcgaggata cacaaaaggc ggtccaggac acgaagaacc
      481 agtacctatg tctacatatg gatggaacat agttaagtta atgagcggaa aggatgtgtt
      541 ctatctccca cctgaaaagt gtgataccct gttgtgtgac attggagaat cttcaccaag
      601 cccaacagtg gaagagagca gaactataag agttttgaag atggttgaac catggctaaa
      661 aaacaaccag ttttgcatta aagttttgaa cccttacatg ccaactgtga ttgagcacct
      721 agaaagacta caaaggaaac atggaggaat gcttgtgaga aatccacttt cacgaaactc
      781 cacgcacgaa atgtactgga tatctaatgg cacaggcaac attgtctctt cagtcaacat
      841 ggtgtctaga ttgctactga acaggttcac gatgacacac aggagaccca ccatagagaa
      901 agatgtggat ttaggagcag gaactcgaca tgttaatgcg gaaccagaaa cacccaacat
      961 ggatgtcatt ggggaaagaa taaaaaggat caaggaggag cataattcaa catggcacta
     1021 tgatgacgaa aacccctaca aaacgtgggc ttaccatgga tcctatgaag tcaaagccac
     1081 aggctcagcc tcctccatga taaatggagt cgtgaaactc ctcaccaaac catgggatgt
     1141 ggtgcccatg gtgacacaga tggcaatgac agacacaact ccatttggcc agcagagagt
     1201 ctttaaagag aaagtggaca ccaggacgcc caggcccctg ccagggacaa gaaaggttat
     1261 ggagatcaca gcggagtggc tctggagaac cctgggaagg aacaaaagac ccagattatg
     1321 cacaagggaa gagtttacaa aaaaggtcag aactaacgca gccatgggcg ccgttttcac
     1381 agaggagaac caatgggaca gtgcgaaagc tgctgttgag gatgaagagt tttggaaact
     1441 tgtggacaga gaacgtgaac tccacaaatt gggcaaatgt ggaagctgcg tttataacat
     1501 gatgggcaag agagagaaaa aactcggaga gtttggcaaa gcaaaaggca gtagagctac
     1561 atggtacatg tggttgggag ccaggtacct tgagttcgaa gcccttggat tcttaaatga
     1621 agaccactgg ttctcgcgtg aaaactctta cagtggagta gaaggagaag gactgcacaa
     1681 gctaggctac atattaaggg acatttccaa gatacccgga ggagccatgt atgctgatga
     1741 cacagctggt tgggacacaa gaataacaga agatgacctg cacaatgagg aaaagatcat
     1801 acagcaaatg gaccctgaac acaggcagtt agcgaacgct atattcaagc tcacatacca
     1861 aaacaaagtg gtcaaagttc aacgaccgac tccaacgggc acggtaatgg atattatatc
     1921 taggaaagac caaaggggca gtggacaagt gggaacttat ggcctgaata cattcaccaa
     1981 catggaagcc cagctagtca gacaaatgga aggagaaggt gtgctgacaa aggcagacct
     2041 cgagaaccct catctgctag agaagaaaat tacacaatgg ttggaaacca aaggagtgga
     2101 gaggttaaaa agaatggcca ttagcgggga tgattgcgtg gtgaaaccaa tcgatgacag
     2161 gttcgctaac gccctgctag ctctgaacga tatgggaaag gttcggaaag acatacctca
     2221 atggcagcca tcaaagggat ggcatgattg gcaacaggtt cctttctgct cccaccactt
     2281 tcatgaattg atcatgaaag atggaagaaa gttggtggtt ccctgcagac cccaggacga
     2341 actaatagga agagcaagaa tctcccaagg agcgggatgg agccttagag aaactgcatg
     2401 tctggggaaa gcctacgccc aaatgtggag tctcatgtat tttcacagaa gagatctcag
     2461 attagcatcc aacgccatat gttcagcagt accagtccac tgggttccca caagtagaac
     2521 gacatggtcc attcatgctc accatcagtg gatgactaca gaagacatgc ttactgtttg
     2581 gaacagggtg tggatagagg aaaatccatg gatggaagac aaaactccag ttacaacttg
     2641 ggaaaatgtt ccatatctag ggaagagaga agaccaatgg tgtggatcac ttattggtct
     2701 cacttccaga gcaacctggg cccagaacat acccacagca attcaacagg tgagaagcct
     2761 tataggcaat gaagagttcc tggactacat gccttcaatg aagagattca ggaaggaaga
     2821 ggagtcgtga cagtaaaggt ggatacggat ccgaattcga gctccgtcga caagcttgcg
     2881 gccgcactcg agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag
     2941 gaagctgagt tggctgctgc caccgctgag caataactag cataacccct tggggcctct
     3001 aaacgggtct tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg
     3061 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     3121 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     3181 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     3241 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     3301 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     3361 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     3421 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     3481 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     3541 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa
     3601 ttaattctta gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat
     3661 tatcaatacc atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc
     3721 agttccatag gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa
     3781 tacaacctat taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag
     3841 tgacgactga atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa
     3901 caggccagcc attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc
     3961 gtgattgcgc ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag
     4021 gaatcgaatg caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat
     4081 caggatattc ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc
     4141 atgcatcatc aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca
     4201 gccagtttag tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt
     4261 tcagaaacaa ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt
     4321 gcccgacatt atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta
     4381 atcgcggcct agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac
     4441 tgtttatgta agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg
     4501 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     4561 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     4621 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     4681 ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     4741 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     4801 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     4861 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     4921 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     4981 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     5041 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     5101 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     5161 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     5221 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
     5281 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt
     5341 acgcatctgt gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg
     5401 ccgcatagtt aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc
     5461 ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc
     5521 ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc
     5581 accgaaacgc gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca
     5641 gatgtctgcc tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg
     5701 gcttctgata aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc
     5761 cgtgtaaggg ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct
     5821 cacgatacgg gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca
     5881 actggcggta tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt
     5941 cgttaataca gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg
     6001 gaacataatg gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc
     6061 gaagaccatt catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt
     6121 tcgctcgcgt atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg
     6181 ggtcctcaac gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat
     6241 ggcctgcttc tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc
     6301 gtgcaagatt ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg
     6361 gtcctcgccg aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa
     6421 gaagacagtc ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac
     6481 tgggttgaag gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact
     6541 tacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct
     6601 gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg
     6661 tttttctttt caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag
     6721 agagttgcag caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg
     6781 tggttaacgg cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga
     6841 tatccgcacc aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct
     6901 gatcgttggc aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt
     6961 gttgaaaacc ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat
     7021 tgcgagtgag atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg
     7081 ggcccgctaa cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc
     7141 gcgtaccgtc ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa
     7201 gaaataacgc cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca
     7261 gcggatagtt aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt
     7321 tacaggcttc gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat
     7381 cggcgcgaga tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg
     7441 tggcaacgcc aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa
     7501 tgtaattcag ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc
     7561 tggcctggtt caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat
     7621 cgtataacgt tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc
     7681 atgccatacc gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc
     7741 ttatgcgact cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc
     7801 gccgcaagga atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct
     7861 gccaccatac ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc
     7921 ccatcggtga tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg
     7981 gccacgatgc gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
