LOCUS       sequence-311251-        6785 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  DENV4 47 peptides from NS2B cofactor linked to NS3 full length by 
            VKTQR linker L30SF31S 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 L30SF31S
FEATURES             Location/Qualifiers
     source          1..6785
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    77..93
                     /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     82..104
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     CDS             126..143
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             165..185
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2322..2339)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2492..2578
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2670..2697
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2716..2807
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2808..2876,2877..3668)
                     /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(3026..3045)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3839..4427
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4328..4347
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4581..4598
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4675..5103)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4785..4806)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4997..5016
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(5141..5160)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5333..5355
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        5433..5510
                     /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             5511..6593
                     /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(5530..5549)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 acatcataac ggttctggca aatattctga aatgagctgt tgacaattaa tcatccggtc
       61 cgtataatct gtggaattgt gagcggataa caatttcaca caggaaacag accatgtcgt
      121 actaccatca ccatcaccat cacgattacg atatcccaac gaccgaaaac ctgtattttc
      181 agggcgccat gggatccgca gacctgtcac tagagaaggc cgccaatgtg cagtgggatg
      241 agatggcaga cataacaggc tcaagcccaa tcatagaagt gaagcaggat gaagatggct
      301 ctttctccat acgggacatc gaggaaacca acatgatagc acaagtgaaa acacaaagat
      361 caggagctct gtgggacgtc ccctcacccg ctgccactca gaaagccaca ctgtcggaag
      421 gagtgtatag gatcatgcaa agagggtctt ctgggaaaac tcaggttgga gtagggatac
      481 acatggaagg tgtattccac acaatgtggc acgtgacaag aggatcagtg atctgccatg
      541 agacagggag attagagcca tcttgggctg acgtcaggaa tgacatgata tcatacggtg
      601 ggggatggag gctcggagac aaatgggaca aagaagaaga tgttcaggtc ctagccatag
      661 aaccaggaaa aaatcctaaa catgtccaaa cgaaaccagg ccttttcaag accctaactg
      721 gagaaattgg agcagtaact ctggatttca aacccggaac gtctggctct cctatcatta
      781 acaagaaagg gaaagttatt ggactctatg gaaatggagt agttaccaaa tcaggtgatt
      841 acgtcagtgc cataacgcaa gctgaaagaa ttggtgagcc agattatgaa gtggatgagg
      901 acatttttcg aaagaaaaga ttaaccataa tggacttaca ccccggagct ggaaagacaa
      961 aaagaattct cccatcaata gtcagagaag ctttaaaaag gaggctgcga actttgattt
     1021 tggctcctac gagagtggtg gcagccgaga tggaagaggc cctacgcgga ctgccaatcc
     1081 gttatcagac cccagctgtg aaatcagatc acacaggaag agagattgtg gacctcatgt
     1141 gtcatgcaac cttcacaacg agacttttgt catcaaccag ggttccaaat tataacctca
     1201 tagtgatgga tgaagcacat ttcactgacc cttgtagtgt cgcggctaga ggatacattt
     1261 caaccagggt ggaaatggga gaggcagcag ccatcttcat gactgcaacc cctcctggat
     1321 cgatagatcc cttcccccag agcaacagcc cgatagaaga catcgaaagg gaaatcccag
     1381 aaaggtcatg gaacacaggg ttcgactgga taacagacta ccaagggaaa actgtgtggt
     1441 ttgttcccag cataaaagct ggaaatgaca ttgcaaattg tttgcgaaag tcgggaaaga
     1501 gagtgatcca gttgagcagg aaaacctttg acacagagta tccaaaaacg aaactcacgg
     1561 actgggattt tgtggtaacc acagacatat ctgaaatggg ggccaatttc agagctggga
     1621 gagtgataga ccccaggaga tgcctcaagc cagttatcct aacagatggg ccagagagag
     1681 ttattctagc aggtccaatc ccagtaactc cagcaagtgc cgctcagaga agagggcgaa
     1741 taggtaggaa tccagcacaa gaagatgacc aatacgtttt ctccggagac ccactaaaga
     1801 atgatgaaga tcatgcccac tggacagaag cgaagatgtt gcttgacaat atctacaccc
     1861 ctgaagggat aattccaaca ttgtttggtc cggaaaggga aaaaacccaa gccattgatg
     1921 gagagtttcg cctcagaggg gaacaaagga agacttttgt ggaattaatg aggagaggag
     1981 accttccggt gtggctgagc tacaaggtag cttctgccgg tatctcttac aaagaccgag
     2041 aatggtgctt cacaggggaa aggaacaacc aaattttaga agaaaacatg gaggttgaaa
     2101 tctggactag agagggagaa aagaaaaaac taaggccaaa atggttagat gcacgtgtgt
     2161 acgctgaccc catggctttg aaggatttca aggagtttgc cagtggaaga aagtaagtcg
     2221 acgagctcac tagtcgcggc cgctttcgaa tctagagcct gcagtctcga ggcatgcggt
     2281 accaagcttg gctgttttgg cggatgagag aagattttca gcctgataca gattaaatca
     2341 gaacgcagaa gcggtctgat aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca
     2401 cctgacccca tgccgaactc agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct
     2461 ccccatgcga gagtagggaa ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga
     2521 ctgggccttt cgttttatct gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc
     2581 gccgggagcg gatttgaacg ttgcgaagca acggcccgga gggtggcggg caggacgccc
     2641 gccataaact gccaggcatc aaattaagca gaaggccatc ctgacggatg gcctttttgc
     2701 gtttctacaa actcttttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     2761 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac
     2821 atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc
     2881 cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca
     2941 tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc
     3001 caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt gttgacgccg
     3061 ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac
     3121 cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca
     3181 taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg
     3241 agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac
     3301 cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg
     3361 caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat
     3421 taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg
     3481 ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg
     3541 cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc
     3601 aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc
     3661 attggtaact gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt
     3721 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     3781 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     3841 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     3901 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     3961 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     4021 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     4081 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     4141 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     4201 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     4261 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     4321 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     4381 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     4441 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     4501 tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc
     4561 gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcctgatgc
     4621 ggtattttct ccttacgcat ctgtgcggta tttcacaccg cataattttg ttaaaattcg
     4681 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4741 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4801 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4861 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4921 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4981 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     5041 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     5101 cgtcccattc gccattcagg ctgctatggt gcactctcag tacaatctgc tctgatgccg
     5161 catagttaag ccagtaccag tcacgtagcg atatcgctac gtgactggta cactccgcta
     5221 tcgctacgtg actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc
     5281 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     5341 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg
     5401 cgcgcgaagg cgaagcggca tgcatttacg ttgacaccat cgaatggtgc aaaacctttc
     5461 gcggtatggc atgatagcgc ccggaagaga gtcaattcag ggtggtgaat gtgaaaccag
     5521 taacgttata cgatgtcgca gagtatgccg gtgtctctta tcagaccgtt tcccgcgtgg
     5581 tgaaccaggc cagccacgtt tctgcgaaaa cgcgggaaaa agtggaagcg gcgatggcgg
     5641 agctgaatta cattcccaac cgcgtggcac aacaactggc gggcaaacag tcgttgctga
     5701 ttggcgttgc cacctccagt ctggccctgc acgcgccgtc gcaaattgtc gcggcgatta
     5761 aatctcgcgc cgatcaactg ggtgccagcg tggtggtgtc gatggtagaa cgaagcggcg
     5821 tcgaagcctg taaagcggcg gtgcacaatc ttctcgcgca acgcgtcagt gggctgatca
     5881 ttaactatcc gctggatgac caggatgcca ttgctgtgga agctgcctgc actaatgttc
     5941 cggcgttatt tcttgatgtc tctgaccaga cacccatcaa cagtattatt ttctcccatg
     6001 aagacggtac gcgactgggc gtggagcatc tggtcgcatt gggtcaccag caaatcgcgc
     6061 tgttagcggg cccattaagt tctgtctcgg cgcgtctgcg tctggctggc tggcataaat
     6121 atctcactcg caatcaaatt cagccgatag cggaacggga aggcgactgg agtgccatgt
     6181 ccggttttca acaaaccatg caaatgctga atgagggcat cgttcccact gcgatgctgg
     6241 ttgccaacga tcagatggcg ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg
     6301 ttggtgcgga tatctcggta gtgggatacg acgataccga agacagctca tgttatatcc
     6361 cgccgttaac caccatcaaa caggattttc gcctgctggg gcaaaccagc gtggaccgct
     6421 tgctgcaact ctctcagggc caggcggtga agggcaatca gctgttgccc gtctcactgg
     6481 tgaaaagaaa aaccaccctg gcacccaata cgcaaaccgc ctctccccgc gcgttggccg
     6541 attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
     6601 gcaattaatg taagttagcg cgaattgatc tggtttgaca gcttatcatc gactgcacgg
     6661 tgcaccaatg cttctggcgt caggcagcca tcggaagctg tggtatggct gtgcaggtcg
     6721 taaatcactg cataattcgt gtcgctcaag gcgcactccc gttctggata atgttttttg
     6781 cgccg
//
