LOCUS       sequence-311249-        6785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DENV4 47 peptides from NS2B cofactor linked to NS3 full length by 
            VKTQR linker S135A mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6785)
  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 6785)
  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: eNS2B47NS3 S135A
FEATURES             Location/Qualifiers
     source          1..6785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    58..74
                     /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     63..85
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             107..124
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             146..166
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2303..2320)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2473..2559
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2651..2678
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2697..2788
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2789..2857,2858..3649)
                     /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(3007..3026)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3820..4408
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4309..4328
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4562..4579
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4656..5084)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4766..4787)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4978..4997
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5122..5141)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5314..5336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5414..5491
                     /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             5492..6574
                     /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(5511..5530)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaatattctg aaatgagctg ttgacaatta atcatccggt ccgtataatc tgtggaattg
       61 tgagcggata acaatttcac acaggaaaca gaccatgtcg tactaccatc accatcacca
      121 tcacgattac gatatcccaa cgaccgaaaa cctgtatttt cagggcgcca tgggatccgc
      181 agacctgtca ctagagaagg ccgccaatgt gcagtgggat gagatggcag acataacagg
      241 ctcaagccca atcatagaag tgaagcagga tgaagatggc tctttctcca tacgggacat
      301 cgaggaaacc aacatgatag cacaagtgaa aacacaaaga tcaggagctc tgtgggacgt
      361 cccctcaccc gctgccactc agaaagccac actgtctgaa ggagtgtata ggatcatgca
      421 aagagggttg tttgggaaaa ctcaggttgg agtagggata cacatggaag gtgtattcca
      481 cacaatgtgg cacgtgacaa gaggatcagt gatctgccat gagacaggga gattagagcc
      541 atcttgggct gacgtcagga atgacatgat atcatacggt ggaggatgga ggctcggaga
      601 caaatgggac aaagaagaag atgttcaggt cctagccata gaaccaggaa aaaatcctaa
      661 acatgtccaa acgaaaccag gccttttcaa gaccctaact ggagaaattg gagcagtaac
      721 tctggatttc aaacccggaa cggcgggctc tcctatcatt aacaagaaag ggaaagttat
      781 tggactctat ggaaatggag tagttaccaa atcaggtgat tacgtcagtg ccataacgca
      841 agctgaaaga attggtgagc cagattatga agtggatgag gacatttttc gaaagaaaag
      901 attaaccata atggacttac accccggagc tggaaagaca aaaagaattc tcccatcaat
      961 agtcagagaa gctttaaaaa ggaggctgcg aactttgatt ttggctccta cgagagtggt
     1021 ggcagccgag atggaagagg ccctacgcgg actgccaatc cgttatcaga ccccagctgt
     1081 gaaatcagat cacacaggaa gagagattgt ggacctcatg tgtcatgcaa ccttcacaac
     1141 gagacttttg tcatcaacca gggttccaaa ttataacctc atagtgatgg atgaagcaca
     1201 tttcactgac ccttgtagtg tcgcggctag aggatacatt tcaaccaggg tggaaatggg
     1261 agaggcagca gccatcttca tgactgcaac ccctcctgga tcgatagatc ccttccccca
     1321 gagcaacagc ccgatagaag acatcgaaag ggaaatccca gaaaggtcat ggaacacagg
     1381 gttcgactgg ataacagact accaagggaa aactgtgtgg tttgttccca gcataaaagc
     1441 tggaaatgac attgcaaatt gtttgcgaaa gtcgggaaag agagtgatcc agttgagcag
     1501 gaaaaccttt gacacagagt atccaaaaac gaaactcacg gactgggatt ttgtggtaac
     1561 cacagacata tctgaaatgg gggccaattt cagagctggg agagtgatag accccaggag
     1621 atgcctcaag ccagttatcc taacagatgg gccagagaga gttattctag caggtccaat
     1681 cccagtaact ccagcaagtg ccgctcagag aagagggcga ataggtagga atccagcaca
     1741 agaagatgac caatacgttt tctccggaga cccactaaag aatgatgaag atcatgccca
     1801 ctggacagaa gcgaagatgt tgcttgacaa tatctacacc cctgaaggga taattccaac
     1861 attgtttggt ccggaaaggg aaaaaaccca agccattgat ggagagtttc gcctcagagg
     1921 ggaacaaagg aagacttttg tggaattaat gaggagagga gaccttccgg tgtggctgag
     1981 ctacaaggta gcttctgccg gtatctctta caaagaccga gaatggtgct tcacagggga
     2041 aaggaacaac caaattttag aagaaaacat ggaggttgaa atctggacta gagagggaga
     2101 aaagaaaaaa ctaaggccaa aatggttaga tgcacgtgtg tacgctgacc ccatggcttt
     2161 gaaggatttc aaggagtttg ccagtggaag aaagtaagtc gacgagctca ctagtcgcgg
     2221 ccgctttcga atctagagcc tgcagtctcg aggcatgcgg taccaagctt ggctgttttg
     2281 gcggatgaga gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga
     2341 taaaacagaa tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact
     2401 cagaagtgaa acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga
     2461 actgccaggc atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc
     2521 tgttgtttgt cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac
     2581 gttgcgaagc aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat
     2641 caaattaagc agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg
     2701 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     2761 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     2821 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     2881 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     2941 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3001 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     3061 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3121 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3181 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3241 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3301 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3361 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     3421 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     3481 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     3541 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     3601 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     3661 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     3721 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3781 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3841 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3901 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3961 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4021 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4081 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4141 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4201 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4261 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4321 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4381 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     4441 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     4501 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     4561 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     4621 tctgtgcggt atttcacacc gcataatttt gttaaaattc gcgttaaatt tttgttaaat
     4681 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     4741 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     4801 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     4861 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     4921 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     4981 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     5041 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcccatt cgccattcag
     5101 gctgctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtacca
     5161 gtcacgtagc gatatcgcta cgtgactggt acactccgct atcgctacgt gactgggtca
     5221 tggctgcgcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     5281 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     5341 caccgtcatc accgaaacgc gcgaggcagc agatcaattc gcgcgcgaag gcgaagcggc
     5401 atgcatttac gttgacacca tcgaatggtg caaaaccttt cgcggtatgg catgatagcg
     5461 cccggaagag agtcaattca gggtggtgaa tgtgaaacca gtaacgttat acgatgtcgc
     5521 agagtatgcc ggtgtctctt atcagaccgt ttcccgcgtg gtgaaccagg ccagccacgt
     5581 ttctgcgaaa acgcgggaaa aagtggaagc ggcgatggcg gagctgaatt acattcccaa
     5641 ccgcgtggca caacaactgg cgggcaaaca gtcgttgctg attggcgttg ccacctccag
     5701 tctggccctg cacgcgccgt cgcaaattgt cgcggcgatt aaatctcgcg ccgatcaact
     5761 gggtgccagc gtggtggtgt cgatggtaga acgaagcggc gtcgaagcct gtaaagcggc
     5821 ggtgcacaat cttctcgcgc aacgcgtcag tgggctgatc attaactatc cgctggatga
     5881 ccaggatgcc attgctgtgg aagctgcctg cactaatgtt ccggcgttat ttcttgatgt
     5941 ctctgaccag acacccatca acagtattat tttctcccat gaagacggta cgcgactggg
     6001 cgtggagcat ctggtcgcat tgggtcacca gcaaatcgcg ctgttagcgg gcccattaag
     6061 ttctgtctcg gcgcgtctgc gtctggctgg ctggcataaa tatctcactc gcaatcaaat
     6121 tcagccgata gcggaacggg aaggcgactg gagtgccatg tccggttttc aacaaaccat
     6181 gcaaatgctg aatgagggca tcgttcccac tgcgatgctg gttgccaacg atcagatggc
     6241 gctgggcgca atgcgcgcca ttaccgagtc cgggctgcgc gttggtgcgg atatctcggt
     6301 agtgggatac gacgataccg aagacagctc atgttatatc ccgccgttaa ccaccatcaa
     6361 acaggatttt cgcctgctgg ggcaaaccag cgtggaccgc ttgctgcaac tctctcaggg
     6421 ccaggcggtg aagggcaatc agctgttgcc cgtctcactg gtgaaaagaa aaaccaccct
     6481 ggcacccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
     6541 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtaagttagc
     6601 gcgaattgat ctggtttgac agcttatcat cgactgcacg gtgcaccaat gcttctggcg
     6661 tcaggcagcc atcggaagct gtggtatggc tgtgcaggtc gtaaatcact gcataattcg
     6721 tgtcgctcaa ggcgcactcc cgttctggat aatgtttttt gcgccgacat cataacggtt
     6781 ctggc
//
