LOCUS       sequence-318477-        7469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  co-expression of DENV4 47 peptides from NS2B cofactor and NS3 full 
            length.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7469)
  AUTHORS   Phoo WW, El Sahili A, Zhang Z, Chen MW, Liew CW, Lescar J, Vasudevan
            SG, Luo D
  TITLE     Crystal structures of full length DENV4 NS2B-NS3 reveal the dynamic 
            interaction between NS2B and NS3.
  JOURNAL   Antiviral Res. 2020 Oct;182:104900. doi: 
            10.1016/j.antiviral.2020.104900. Epub 2020 Aug 5.
  PUBMED    32763315
REFERENCE   2  (bases 1 to 7469)
  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: "Antiviral 
            Res."; date: "2020-10"; volume: "182:104900. doi"; pages: " 
            10.1016/j.antiviral.2020.104900. Epub 2020 Aug 5"
COMMENT     Sequence Label: bNS2B47NS3
FEATURES             Location/Qualifiers
     source          1..7469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     116..135
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        219..237
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    238..262
                     /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             277..299
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             310..327
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..372
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     promoter        560..578
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    579..603
                     /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)."
     CDS             2552..2569
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2622..2640)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2636..2683
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2720..3175
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2807..2826)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3017..3038
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3267..4082)
                     /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(3267..4082)
                     /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     3990..4009
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4074..4093
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4204..4792
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4693..4712
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4946..4963
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4978..5117
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5043..5062)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5203..5225
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5219..5410)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6219..7301)
                     /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     7263..7282
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7302..7379)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
       61 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      121 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      181 gtccggcgta gaggatcgag atctcgatcc cgcgaaatta atacgactca ctatagggga
      241 attgtgagcg gataacaatt cccctctaga aataattttg tttaacttta agaaggagat
      301 atacatatgc accatcatca tcatcattct tctggtgtag atctgggtac cgagaacctg
      361 tacttccaat ccatgggtag cgctgacctg agcctggaaa aagctgctaa cgtgcagtgg
      421 gatgagatgg ctgatatcac cggcagcagc ccgatcatcg aagttaaaca ggatgaagat
      481 ggtagcttca gcatccgtga tgttgaagaa acaaacatga tcacctaaga attcttgtac
      541 acggccgcat aatcgaaatt aatacgactc actatagggg aattgtgagc ggataacaat
      601 tccccatctt agtatattag ttaagtataa gaaggagata tacatatgag cggtgcactg
      661 tgggatgttc cgagcccggc ggcgaccaaa aaagcagctc tgtctgaagg tgtttatcgt
      721 atcatgcagc gtggcctgtt cggcaaaacc caggttggtg ttggtattca catggaaggc
      781 gtttttcaca ccatgtggca tgttacccgt ggtagcgtta tctgccatga aaccggccgt
      841 ctggaaccgt cttgggcgga tgttcgtaat gatatgatta gctacggtgg cggctggcgt
      901 ctgggcgata aatgggacaa agaagaagat gttcaggttc tggcgatcga accgggtaaa
      961 aacccgaaac acgttcagac caaaccgggc ctgttcaaaa ccctgactgg cgaaatcggt
     1021 gcggttaccc tggattttaa accgggtacc tctggttctc cgatcattaa ccgtaaaggc
     1081 aaagttattg gtctgtacgg taatggcgtt gtgaccaaat ctggcgatta tgtttctgct
     1141 attactcagg ctgaacgtat cggtgaaccg gattatgaag ttgatgaaga tatctttcgt
     1201 aaaaaacgtt taaccatcat ggatctgcat ccgggtgcgg gtaaaaccaa acgtatcctg
     1261 ccgtctattg ttcgtgaagc actgaaacgt cgtctgcgta ccctgatctt agcgccgacc
     1321 cgtgttgttg cagctgaaat ggaagaagcg ctgcgcggct tgccgattcg ttatcaaact
     1381 ccggcagtta aatctgaaca caccggccgt gaaatcgttg atctgatgtg tcatgcgacc
     1441 ttcaccaccc gtctgctgag ctccacccgt gttccgaact ataacctgat tgttatggat
     1501 gaagcgcatt tcaccgatcc gtctagcgtt gcagcacgtg gttacatctc tacccgtgtt
     1561 gaaatgggtg aagcggcggc tatcttcatg accgcgaccc cgccgggtgc tactgatccg
     1621 ttcccgcaat ctaactctcc tatcgaagac atcgaacgtg aaattccgga acgctcctgg
     1681 aacactggtt ttgactggat tactgattac cagggtaaaa ccgtttggtt cgttccgtct
     1741 attaaagccg gtaacgacat tgctaactgc ctgcgtaaat ctggcaaaaa agttatccag
     1801 ctgtctcgta aaactttcga tactgaatat cctaaaacca aactgaccga ttgggatttc
     1861 gttgttacta ccgacatctc tgaaatgggt gctaactttc gtgcgggtcg tgttatcgat
     1921 ccacgtcgtt gtctgaaacc ggttattctg ccggatggcc cagaacgtgt tattctggca
     1981 ggtccgatcc cggttacccc tgcttctgca gctcagcgtc gcggtcgtat cggtcgtaac
     2041 ccggcacagg aagatgatca gtacgttttt agcggtgacc cgctgaaaaa cgatgaagat
     2101 cacgctcact ggaccgaagc taaaatgctg ctggataaca tctatacccc agaaggcatt
     2161 attccgactc tgtttggccc ggaacgtgaa aaaacccagg cgatcgatgg tgaattccgt
     2221 ctgcgtggtg aacagcgtaa aacctttgtg gaactgatgc gtcgtggcga tctgccggtt
     2281 tggctgagct acaaagttgc atctgcaggc atcagctatg aagatcgtga atggtgcttc
     2341 accggtgaac gtaacaacca gatcctggaa gaaaacatgg aagttgaaat ctggacccgt
     2401 gaaggtgaaa agaaaaaact gcgtccgcgt tggctggatg ctcgtgttta cgctgatccg
     2461 atggcgctga aagattttaa agaattcgct agcggtcgta aataaggatc cgaattcgag
     2521 ctccgtcgac aagcttgcgg ccgcactcga gcaccaccac caccaccact gagatccggc
     2581 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     2641 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     2701 atccggattg gcgaatggga cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2761 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2821 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2881 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2941 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     3001 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     3061 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     3121 ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac aatttaggtg
     3181 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3241 atatgtatcc gctcatgaat taattcttag aaaaactcat cgagcatcaa atgaaactgc
     3301 aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt ctgtaatgaa
     3361 ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg gtctgcgatt
     3421 ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat aaggttatca
     3481 agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag tttatgcatt
     3541 tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc actcgcatca
     3601 accaaaccgt tattcattcg tgattgcgcc tgagcgagac gaaatacgcg atcgctgtta
     3661 aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc cagcgcatca
     3721 acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt tttcccgggg
     3781 atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt gatggtcgga
     3841 agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac atcattggca
     3901 acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc atacaatcga
     3961 tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc atataaatca
     4021 gcatccatgt tggaatttaa tcgcggccta gagcaagacg tttcccgttg aatatggctc
     4081 ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca tgaccaaaat
     4141 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     4201 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4261 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4321 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     4381 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4441 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4501 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4561 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4621 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4681 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4741 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4801 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4861 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4921 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     4981 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     5041 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     5101 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     5161 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     5221 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     5281 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     5341 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     5401 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catgggggta atgataccga
     5461 tgaaacgaga gaggatgctc acgatacggg ttactgatga tgaacatgcc cggttactgg
     5521 aacgttgtga gggtaaacaa ctggcggtat ggatgcggcg ggaccagaga aaaatcactc
     5581 agggtcaatg ccagcgcttc gttaatacag atgtaggtgt tccacagggt agccagcagc
     5641 atcctgcgat gcagatccgg aacataatgg tgcagggcgc tgacttccgc gtttccagac
     5701 tttacgaaac acggaaaccg aagaccattc atgttgttgc tcaggtcgca gacgttttgc
     5761 agcagcagtc gcttcacgtt cgctcgcgta tcggtgattc attctgctaa ccagtaaggc
     5821 aaccccgcca gcctagccgg gtcctcaacg acaggagcac gatcatgcgc acccgtgggg
     5881 ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg tttggtggcg ggaccagtga
     5941 cgaaggcttg agcgagggcg tgcaagattc cgaataccgc aagcgacagg ccgatcatcg
     6001 tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc
     6061 ctacgagttg catgataaag aagacagtca taagtgcggc gacgatagtc atgccccgcg
     6121 cccaccggaa ggagctgact gggttgaagg ctctcaaggg catcggtcga gatcccggtg
     6181 cctaatgagt gagctaactt acattaattg cgttgcgctc actgcccgct ttccagtcgg
     6241 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     6301 gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag ctgattgccc
     6361 ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg ccccagcagg
     6421 cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc ttcggtatcg
     6481 tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt aatggcgcgc
     6541 attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac gatgccctca
     6601 ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc
     6661 gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag acgcagacgc
     6721 gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc caatgcgacc
     6781 agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact gttgatgggt
     6841 gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc ttccacagca
     6901 atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg ttgcgcgaga
     6961 agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat cgacaccacc
     7021 acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg
     7081 tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc cgccagttgt
     7141 tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac tttttcccgc
     7201 gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg ataagagaca
     7261 ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac cctgaattga
     7321 ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc gatggtgtcc
     7381 gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     7441 gaggccgttg agcaccgccg ccgcaagga
//
