LOCUS       sequence-311234-        6791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DENV4 47 peptides from NS2B cofactor linked to NS3 full length by 
            GGGGSGGGG linker S135A mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6791)
  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 6791)
  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: gNS2B47NS3 S135A
FEATURES             Location/Qualifiers
     source          1..6791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    246..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)."
     primer_bind     251..273
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             295..312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             334..354
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2497..2514)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2667..2753
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2845..2872
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2891..2982
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2983..3051,3052..3843)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3201..3220)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4014..4602
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4503..4522
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4756..4773
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4850..5278)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4960..4981)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5172..5191
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5316..5335)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5508..5530
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5608..5685
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             5686..6768
                     /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     complement(5705..5724)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 agcgcgaatt gatctggttt gacagcttat catcgactgc acggtgcacc aatgcttctg
       61 gcgtcaggca gccatcggaa gctgtggtat ggctgtgcag gtcgtaaatc actgcataat
      121 tcgtgtcgct caaggcgcac tcccgttctg gataatgttt tttgcgccga catcataacg
      181 gttctggcaa atattctgaa atgagctgtt gacaattaat catccggtcc gtataatctg
      241 tggaattgtg agcggataac aatttcacac aggaaacaga ccatgtcgta ctaccatcac
      301 catcaccatc acgattacga tatcccaacg accgaaaacc tgtattttca gggcgccatg
      361 ggatccgcag acctgtcact agagaaggcc gccaatgtgc agtgggatga gatggcagac
      421 ataacaggct caagcccaat catagaagtg aagcaggatg aagatggctc tttctccata
      481 cgggacatcg aggaaaccaa tatgatagga ggaggaggat caggaggagg aggatcagga
      541 gctctgtggg acgtcccctc acccgctgcc actcagaaag ccacactgtc tgaaggagtg
      601 tataggatca tgcaaagagg gttgtttggg aaaactcagg ttggagtagg gatacacatg
      661 gaaggtgtat tccacacaat gtggcacgtg acaagaggat cagtgatctg ccatgagaca
      721 gggagattag agccatcttg ggctgacgtc aggaatgaca tgatatcata cggtggggga
      781 tggaggctcg gagacaaatg ggacaaagaa gaagatgttc aggtcctagc catagaacca
      841 ggaaaaaatc ctaaacatgt ccaaacgaaa ccaggccttt tcaagaccct aactggagaa
      901 attggagcag taactctgga tttcaaaccc ggaacggcgg gctctcctat cattaacaag
      961 aaagggaaag ttattggact ctatggaaat ggagtagtta ccaaatcagg tgattacgtc
     1021 agtgccataa cgcaagctga aagaattggt gagccagatt atgaagtgga tgaggacatt
     1081 tttcgaaaga aaagattaac cataatggac ttacaccccg gagctggaaa gacaaaaaga
     1141 attctcccat caatagtcag agaagcttta aaaaggaggc tgcgaacttt gattttggct
     1201 cctacgagag tggtggcagc cgagatggaa gaggccctac gcggactgcc aatccgttat
     1261 cagaccccag ctgtgaaatc agatcacaca ggaagagaga ttgtggacct catgtgtcat
     1321 gcaaccttca caacgagact tttgtcatca accagggttc caaattataa cctcatagtg
     1381 atggatgaag cacatttcac tgacccttgt agtgtcgcgg ctagaggata catttcaacc
     1441 agggtggaaa tgggagaggc agcagccatc ttcatgactg caacccctcc tggatcgata
     1501 gatcccttcc cccagagcaa cagcccgata gaagacatcg aaagggaaat cccagaaagg
     1561 tcatggaaca cagggttcga ctggataaca gactaccaag ggaaaactgt gtggtttgtt
     1621 cccagcataa aagctggaaa tgacattgca aattgtttgc gaaagtcggg aaagagagtg
     1681 atccagttga gcaggaaaac ctttgacaca gagtatccaa aaacgaaact cacggactgg
     1741 gattttgtgg taaccacaga catatctgaa atgggggcca atttcagagc tgggagagtg
     1801 atagacccca ggagatgcct caagccagtt atcctaacag atgggccaga gagagttatt
     1861 ctagcaggtc caatcccagt aactccagca agtgccgctc agagaagagg gcgaataggt
     1921 aggaatccag cacaagaaga tgaccaatac gttttctccg gagacccact aaagaatgat
     1981 gaagatcatg cccactggac agaagcgaag atgttgcttg acaatatcta cacccctgaa
     2041 gggataattc caacattgtt tggtccggaa agggaaaaaa cccaagccat tgatggagag
     2101 tttcgcctca gaggggaaca aaggaagact tttgtggaat taatgaggag aggagacctt
     2161 ccggtgtggc tgagctacaa ggtagcttct gccggtatct cttacaaaga ccgagaatgg
     2221 tgcttcacag gggaaaggaa caaccaaatt ttagaagaaa acatggaggt tgaaatctgg
     2281 actagagagg gagaaaagaa aaaactaagg ccaaaatggt tagatgcacg tgtgtacgct
     2341 gaccccatgg ctttgaagga tttcaaggag tttgccagtg gaagaaagta agtcgacgag
     2401 ctcactagtc gcggccgctt tcgaatctag agcctgcagt ctcgaggcat gcggtaccaa
     2461 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     2521 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     2581 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     2641 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2701 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     2761 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     2821 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     2881 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2941 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3001 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3061 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3121 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3181 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3241 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3301 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3361 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     3421 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     3481 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     3541 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     3601 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     3661 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     3721 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     3781 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     3841 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     3901 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     3961 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4021 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4081 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4141 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     4201 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     4261 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     4321 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     4381 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     4441 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     4501 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     4561 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     4621 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     4681 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     4741 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat
     4801 tttctcctta cgcatctgtg cggtatttca caccgcataa ttttgttaaa attcgcgtta
     4861 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     4921 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     4981 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     5041 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     5101 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     5161 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     5221 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc
     5281 cattcgccat tcaggctgct atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5341 ttaagccagt accagtcacg tagcgatatc gctacgtgac tggtacactc cgctatcgct
     5401 acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg
     5461 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     5521 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca attcgcgcgc
     5581 gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac ctttcgcggt
     5641 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     5701 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     5761 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     5821 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     5881 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     5941 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     6001 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     6061 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     6121 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     6181 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     6241 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     6301 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     6361 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     6421 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     6481 gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     6541 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     6601 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     6661 agaaaaacca ccctggcacc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     6721 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     6781 taatgtaagt t
//
