LOCUS       sequence-251110-        7009 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CHIKV nsP2h (1-465 aa) in bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7009)
  AUTHORS   Law YS, Utt A, Tan YB, Zheng J, Wang S, Chen MW, Griffin PR, Merits 
            A, Luo D
  TITLE     Structural insights into RNA recognition by the Chikungunya virus 
            nsP2 helicase.
  JOURNAL   Proc Natl Acad Sci U S A. 2019 May 7;116(19):9558-9567. doi: 
            10.1073/pnas.1900656116. Epub 2019 Apr 18.
  PUBMED    31000599
REFERENCE   2  (bases 1 to 7009)
  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"; doi: 
            "10.1073/pnas.1900656116"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2019-05-7- 7"; volume: "116"; issue: "19"; pages: "9558-9567"
COMMENT     Sequence Label: pSUMO-LIC-CHIKV-nsP2h
FEATURES             Location/Qualifiers
     source          1..7009
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     54..73
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        54..72
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    73..97
                     /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             112..134
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             154..171
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1882..1926
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1927..1944
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1997..2015)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2011..2058
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2095..2550
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2182..2201)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2392..2413
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2642..3457)
                     /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(2642..3457)
                     /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     3365..3384
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3449..3468
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3579..4167
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4068..4087
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4321..4338
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4353..4492
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4418..4437)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4578..4600
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4594..4785)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5594..6676)
                     /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     6638..6657
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6677..6754)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6960..6979
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 tgccggccac gatgcgtccg gcgtagagga tcgagatctc gatcccgcga aattaatacg
       61 actcactata ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa
      121 ctttaagaag gagatataca tatgtgcagc agccatcatc atcatcatca cggcagcggc
      181 agcggctctg accaggaggc aaaaccttca actgaggact tgggggataa gaaggaaggt
      241 gaatatatta aactcaaagt cattggacag gatagcagtg agattcactt caaagtgaaa
      301 atgacaacac atctcaagaa actcaaagaa tcatactgtc aaagacaggg tgttccaatg
      361 aattcactca ggtttctctt tgagggtcag agaattgctg ataatcatac tccaaaagaa
      421 ctgggaatgg aggaagaaga tgtgattgaa gtttaccagg agcaaacggg aggtggaata
      481 atagagactc cgagaggagc tatcaaagtt actgcccaac caacagacca cgtcgtggga
      541 gagtacctgg tactctcccc gcagaccgta ctacgtagcc agaagctcag tctgattcac
      601 gctttggcgg agcaagtgaa gacgtgcacg cacaacggac gagcagggag gtatgcggtc
      661 gaagcgtacg acggccgagt cctagtgccc tcaggctatg caatctcgcc tgaagacttc
      721 cagagtctaa gcgaaagcgc aacgatggtg tataacgaaa gagagttcgt aaacagaaag
      781 ctacaccata ttgcgatgca cggaccagcc ctgaacaccg acgaagagtc gtatgagctg
      841 gtgagggcag agaggacaga acacgagtac gtctacgacg tggatcagag aagatgctgt
      901 aagaaggaag aagccgcagg actggtactg gtgggcgact tgactaatcc gccctaccac
      961 gaattcgcat atgaagggct aaaaatccgc cctgcctgcc catacaaaat tgcagtcata
     1021 ggagtcttcg gagtaccggg atctggcaag tcagctatta tcaagaacct agttaccagg
     1081 caggacctgg tgactagcgg aaagaaagaa aactgccaag aaatcaccac cgacgtgatg
     1141 agacagagag gtctagagat atctgcacgt acggttgact cgctgctctt gaatggatgc
     1201 aacagaccag tcgacgtgtt gtacgtagac gaggcgtttg cgtgccactc tggaacgcta
     1261 cttgctttga tcgccttggt gagaccaagg cagaaagttg tactttgtgg tgacccgaag
     1321 cagtgcggct tcttcaatat gatgcagatg aaagtcaact ataatcacaa catctgcacc
     1381 caagtgtacc acaaaagtat ctccaggcgg tgtacactgc ctgtgaccgc cattgtgtca
     1441 tcgttgcatt acgaaggcaa aatgcgcact acgaatgagt acaacaagcc gattgtagtg
     1501 gacactacag gctcaacaaa acctgaccct ggagacctcg tgttaacgtg cttcagaggg
     1561 tgggttaaac aactgcaaat tgactatcgt ggatacgagg tcatgacagc agccgcatcc
     1621 caagggttaa ccagaaaagg agtttacgca gttagacaaa aagttaatga aaacccgctc
     1681 tatgcatcaa cgtcagagca cgtcaacgta ctcctaacgc gtacggaagg taaactggta
     1741 tggaagacac tttccggcga cccgtggata aagacgctgc agaacccacc gaaaggaaac
     1801 ttcaaagcaa ctattaagga gtgggaggtg gagcatgcat caataatggc gggcatctgc
     1861 agtcaccaat aaagcggtgg cggtctgaac gacatcttcg aggctcagaa aatcgaatgg
     1921 cacgaacacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     1981 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     2041 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     2101 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     2161 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     2221 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     2281 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc
     2341 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     2401 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga
     2461 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     2521 attttaacaa aatattaacg cttacaattt aggtggcact tttcggggaa atgtgcgcgg
     2581 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgaattaatt
     2641 cttagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa
     2701 taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc
     2761 ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac
     2821 ctattaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga
     2881 ctgaatccgg tgagaatggc aaaagtttat gcatttcttt ccagacttgt tcaacaggcc
     2941 agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt
     3001 gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg
     3061 aatgcaaccg gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat
     3121 attcttctaa tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat
     3181 catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt
     3241 ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa
     3301 acaactctgg cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga
     3361 cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg
     3421 gcctagagca agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta
     3481 tgtaagcaga cagttttatt gttcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3541 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3601 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3661 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3721 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3781 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3841 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3901 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3961 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4021 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4081 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4141 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4201 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4261 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4321 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     4381 ctgtgcggta tttcacaccg caatggtgca ctctcagtac aatctgctct gatgccgcat
     4441 agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg cgccccgaca
     4501 cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag
     4561 acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa
     4621 acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg tgaagcgatt cacagatgtc
     4681 tgcctgttca tccgcgtcca gctcgttgag tttctccaga agcgttaatg tctggcttct
     4741 gataaagcgg gccatgttaa gggcggtttt ttcctgtttg gtcactgatg cctccgtgta
     4801 agggggattt ctgttcatgg gggtaatgat accgatgaaa cgagagagga tgctcacgat
     4861 acgggttact gatgatgaac atgcccggtt actggaacgt tgtgagggta aacaactggc
     4921 ggtatggatg cggcgggacc agagaaaaat cactcagggt caatgccagc gcttcgttaa
     4981 tacagatgta ggtgttccac agggtagcca gcagcatcct gcgatgcaga tccggaacat
     5041 aatggtgcag ggcgctgact tccgcgtttc cagactttac gaaacacgga aaccgaagac
     5101 cattcatgtt gttgctcagg tcgcagacgt tttgcagcag cagtcgcttc acgttcgctc
     5161 gcgtatcggt gattcattct gctaaccagt aaggcaaccc cgccagccta gccgggtcct
     5221 caacgacagg agcacgatca tgcgcacccg tggggccgcc atgccggcga taatggcctg
     5281 cttctcgccg aaacgtttgg tggcgggacc agtgacgaag gcttgagcga gggcgtgcaa
     5341 gattccgaat accgcaagcg acaggccgat catcgtcgcg ctccagcgaa agcggtcctc
     5401 gccgaaaatg acccagagcg ctgccggcac ctgtcctacg agttgcatga taaagaagac
     5461 agtcataagt gcggcgacga tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     5521 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     5581 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5641 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     5701 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     5761 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     5821 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatatccg
     5881 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     5941 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     6001 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     6061 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     6121 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     6181 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     6241 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     6301 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     6361 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     6421 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     6481 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     6541 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     6601 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     6661 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     6721 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     6781 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     6841 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     6901 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     6961 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtga
//
