LOCUS       sequence-311228-        6791 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DENV4 47 peptides from NS2B cofactor linked to NS3 full length by 
            GGGGSGGGG linker L30SF31S 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 L30SF31S
FEATURES             Location/Qualifiers
     source          1..6791
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    153..169
                     /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     158..180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             202..219
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             241..261
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2404..2421)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2574..2660
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2752..2779
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2798..2889
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2890..2958,2959..3750)
                     /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(3108..3127)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3921..4509
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4410..4429
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4663..4680
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4757..5185)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4867..4888)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5079..5098
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5223..5242)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5415..5437
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5515..5592
                     /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             5593..6675
                     /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(5612..5631)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 tgtgcaggtc gtaaatcact gcataattcg tgtcgctcaa ggcgcactcc cgttctggat
       61 aatgtttttt gcgccgacat cataacggtt ctggcaaata ttctgaaatg agctgttgac
      121 aattaatcat ccggtccgta taatctgtgg aattgtgagc ggataacaat ttcacacagg
      181 aaacagacca tgtcgtacta ccatcaccat caccatcacg attacgatat cccaacgacc
      241 gaaaacctgt attttcaggg cgccatggga tccgcagacc tgtcactaga gaaggccgcc
      301 aatgtgcagt gggatgagat ggcagacata acaggctcaa gcccaatcat agaagtgaag
      361 caggatgaag atggctcttt ctccatacgg gacatcgagg aaaccaatat gataggagga
      421 ggaggatcag gaggaggagg atcaggagct ctgtgggacg tcccctcacc cgctgccact
      481 cagaaagcca cactgtctga aggagtgtat aggatcatgc aaagagggtc ttctgggaaa
      541 actcaggttg gagtagggat acacatggaa ggtgtattcc acacaatgtg gcacgtgaca
      601 agaggatcag tgatctgcca tgagacaggg agattagagc catcttgggc tgacgtcagg
      661 aatgacatga tatcatacgg tgggggatgg aggctcggag acaaatggga caaagaagaa
      721 gatgttcagg tcctagccat agaaccagga aaaaatccta aacatgtcca aacgaaacca
      781 ggccttttca agaccctaac tggagaaatt ggagcagtaa ctctggattt caaacccgga
      841 acgtctggct ctcctatcat taacaagaaa gggaaagtta ttggactcta tggaaatgga
      901 gtagttacca aatcaggtga ttacgtcagt gccataacgc aagctgaaag aattggtgag
      961 ccagattatg aagtggatga ggacattttt cgaaagaaaa gattaaccat aatggactta
     1021 caccccggag ctggaaagac aaaaagaatt ctcccatcaa tagtcagaga agctttaaaa
     1081 aggaggctgc gaactttgat tttggctcct acgagagtgg tggcagccga gatggaagag
     1141 gccctacgcg gactgccaat ccgttatcag accccagctg tgaaatcaga tcacacagga
     1201 agagagattg tggacctcat gtgtcatgca accttcacaa cgagactttt gtcatcaacc
     1261 agggttccaa attataacct catagtgatg gatgaagcac atttcactga cccttgtagt
     1321 gtcgcggcta gaggatacat ttcaaccagg gtggaaatgg gagaggcagc agccatcttc
     1381 atgactgcaa cccctcctgg atcgatagat cccttccccc agagcaacag cccgatagaa
     1441 gacatcgaaa gggaaatccc agaaaggtca tggaacacag ggttcgactg gataacagac
     1501 taccaaggga aaactgtgtg gtttgttccc agcataaaag ctggaaatga cattgcaaat
     1561 tgtttgcgaa agtcgggaaa gagagtgatc cagttgagca ggaaaacctt tgacacagag
     1621 tatccaaaaa cgaaactcac ggactgggat tttgtggtaa ccacagacat atctgaaatg
     1681 ggggccaatt tcagagctgg gagagtgata gaccccagga gatgcctcaa gccagttatc
     1741 ctaacagatg ggccagagag agttattcta gcaggtccaa tcccagtaac tccagcaagt
     1801 gccgctcaga gaagagggcg aataggtagg aatccagcac aagaagatga ccaatacgtt
     1861 ttctccggag acccactaaa gaatgatgaa gatcatgccc actggacaga agcgaagatg
     1921 ttgcttgaca atatctacac ccctgaaggg ataattccaa cattgtttgg tccggaaagg
     1981 gaaaaaaccc aagccattga tggagagttt cgcctcagag gggaacaaag gaagactttt
     2041 gtggaattaa tgaggagagg agaccttccg gtgtggctga gctacaaggt agcttctgcc
     2101 ggtatctctt acaaagaccg agaatggtgc ttcacagggg aaaggaacaa ccaaatttta
     2161 gaagaaaaca tggaggttga aatctggact agagagggag aaaagaaaaa actaaggcca
     2221 aaatggttag atgcacgtgt gtacgctgac cccatggctt tgaaggattt caaggagttt
     2281 gccagtggaa gaaagtaagt cgacgagctc actagtcgcg gccgctttcg aatctagagc
     2341 ctgcagtctc gaggcatgcg gtaccaagct tggctgtttt ggcggatgag agaagatttt
     2401 cagcctgata cagattaaat cagaacgcag aagcggtctg ataaaacaga atttgcctgg
     2461 cggcagtagc gcggtggtcc cacctgaccc catgccgaac tcagaagtga aacgccgtag
     2521 cgccgatggt agtgtggggt ctccccatgc gagagtaggg aactgccagg catcaaataa
     2581 aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg tcggtgaacg
     2641 ctctcctgag taggacaaat ccgccgggag cggatttgaa cgttgcgaag caacggcccg
     2701 gagggtggcg ggcaggacgc ccgccataaa ctgccaggca tcaaattaag cagaaggcca
     2761 tcctgacgga tggccttttt gcgtttctac aaactctttt gtttattttt ctaaatacat
     2821 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     2881 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     2941 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3001 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3061 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3121 gtattatccc gtgttgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3181 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3241 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3301 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3361 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3421 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3481 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3541 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3601 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     3661 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     3721 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     3781 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     3841 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     3901 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     3961 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4021 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     4081 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4141 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4201 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4261 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4321 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4381 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4441 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4501 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4561 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4621 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     4681 gaagcggaag agcgcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac
     4741 cgcataattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     4801 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     4861 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     4921 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     4981 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     5041 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     5101 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     5161 cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgctatg gtgcactctc
     5221 agtacaatct gctctgatgc cgcatagtta agccagtacc agtcacgtag cgatatcgct
     5281 acgtgactgg tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     5341 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5401 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5461 cgcgaggcag cagatcaatt cgcgcgcgaa ggcgaagcgg catgcattta cgttgacacc
     5521 atcgaatggt gcaaaacctt tcgcggtatg gcatgatagc gcccggaaga gagtcaattc
     5581 agggtggtga atgtgaaacc agtaacgtta tacgatgtcg cagagtatgc cggtgtctct
     5641 tatcagaccg tttcccgcgt ggtgaaccag gccagccacg tttctgcgaa aacgcgggaa
     5701 aaagtggaag cggcgatggc ggagctgaat tacattccca accgcgtggc acaacaactg
     5761 gcgggcaaac agtcgttgct gattggcgtt gccacctcca gtctggccct gcacgcgccg
     5821 tcgcaaattg tcgcggcgat taaatctcgc gccgatcaac tgggtgccag cgtggtggtg
     5881 tcgatggtag aacgaagcgg cgtcgaagcc tgtaaagcgg cggtgcacaa tcttctcgcg
     5941 caacgcgtca gtgggctgat cattaactat ccgctggatg accaggatgc cattgctgtg
     6001 gaagctgcct gcactaatgt tccggcgtta tttcttgatg tctctgacca gacacccatc
     6061 aacagtatta ttttctccca tgaagacggt acgcgactgg gcgtggagca tctggtcgca
     6121 ttgggtcacc agcaaatcgc gctgttagcg ggcccattaa gttctgtctc ggcgcgtctg
     6181 cgtctggctg gctggcataa atatctcact cgcaatcaaa ttcagccgat agcggaacgg
     6241 gaaggcgact ggagtgccat gtccggtttt caacaaacca tgcaaatgct gaatgagggc
     6301 atcgttccca ctgcgatgct ggttgccaac gatcagatgg cgctgggcgc aatgcgcgcc
     6361 attaccgagt ccgggctgcg cgttggtgcg gatatctcgg tagtgggata cgacgatacc
     6421 gaagacagct catgttatat cccgccgtta accaccatca aacaggattt tcgcctgctg
     6481 gggcaaacca gcgtggaccg cttgctgcaa ctctctcagg gccaggcggt gaagggcaat
     6541 cagctgttgc ccgtctcact ggtgaaaaga aaaaccaccc tggcacccaa tacgcaaacc
     6601 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     6661 gaaagcgggc agtgagcgca acgcaattaa tgtaagttag cgcgaattga tctggtttga
     6721 cagcttatca tcgactgcac ggtgcaccaa tgcttctggc gtcaggcagc catcggaagc
     6781 tgtggtatgg c
//
