LOCUS       sequence-417058-        7430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of Zika NS5 RdRp.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7430)
  TITLE     The SARS-Cov-2 main protease (Mpro) is responsible for viral 
            polyprotein processing and therefore a therapeutic target.
REFERENCE   2  (bases 1 to 7430)
  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"
COMMENT     Sequence Label: Zika NS5 RdRp
FEATURES             Location/Qualifiers
     source          1..7430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             94..111
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             115..441
                     /codon_start=1
                     /gene="trxA"
                     /product="E. coli thioredoxin"
                     /label=TrxA
                     /translation="MSDKIIHLTDDSFDTDVLKADGAILVDFWAEWCGPCKMIAPILDE
                     IADEYQGKLTVAKLNIDQNPGTAPKYGIRGIPTLLLFKNGEVAATKVGALSKGQLKEFL
                     DANLA"
     CDS             460..480
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2295..2312
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2365..2383)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2379..2426
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2463..2918
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2550..2569)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2760..2781
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3010..3825)
                     /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(3010..3825)
                     /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     3733..3752
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3817..3836
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3947..4535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4436..4455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4689..4706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4721..4860
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4786..4805)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4946..4968
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4962..5153)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5962..7044)
                     /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     7006..7025
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7045..7122)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7328..7347
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg aaacatcacc atcaccatca ccccatgagc
      121 gataaaatta ttcacctgac tgacgacagt tttgacacgg atgtactcaa agcggacggg
      181 gcgatcctcg tcgatttctg ggcagagtgg tgcggtccgt gcaaaatgat cgccccgatt
      241 ctggatgaaa tcgctgacga atatcagggc aaactgaccg ttgcaaaact gaacatcgat
      301 caaaaccctg gcactgcgcc gaaatatggc atccgtggta tcccgactct gctgctgttc
      361 aaaaacggtg aagtggcggc aaccaaagtg ggtgcactgt ctaaaggtca gttgaaagag
      421 ttcctcgacg ctaacctggc cggatctggc agtggttctg agaatcttta ttttcagggc
      481 gccatgtacc atgggagcta cgaagccccc acgcaaggat cagcgtcttc cctcgtgaac
      541 ggggttgtta gactcctgtc aaagccttgg gacgtggtga ctggagttac aggaatagcc
      601 atgactgaca ccacaccata cggccaacaa agagtcttca aagaaaaagt ggacaccagg
      661 gtgccagatc cccaagaagg cactcgccag gtaatgaaca tagtctcttc ctggctgtgg
      721 aaggagctgg ggaaacgcaa gcggccacgc gtctgcacca aagaagagtt tatcaacaag
      781 gtgcgcagca atgcagcact gggagcaata tttgaagagg aaaaagaatg gaagacggct
      841 gtggaagctg tgaatgatcc aaggttttgg gccctagtgg atagggagag agaacaccac
      901 ctgagaggag agtgtcacag ctgtgtgtac aacatgatgg gaaaaagaga aaagaagcaa
      961 ggagagttcg ggaaagcaaa aggtagccgc gccatctggt acatgtggtt gggagccaga
     1021 ttcttggagt ttgaagccct tggattcttg aacgaggacc attggatggg aagagaaaac
     1081 tcaggaggtg gagtcgaagg gttaggattg caaagacttg gatacattct agaagaaatg
     1141 aatcgggcac caggaggaaa gatgtacgca gatgacactg ctggctggga cacccgcatt
     1201 agtaagtttg atctggagaa tgaagctctg attaccaacc aaatggagga agggcacaga
     1261 actctggcgt tggccgtgat taaatacaca taccaaaaca aagtggtgaa ggttctcaga
     1321 ccagctgaag gaggaaaaac agttatggac atcatttcaa gacaagacca gagagggagt
     1381 ggacaagttg tcacttatgc tctcaacaca ttcaccaact tggtggtgca gcttatccgg
     1441 aacatggaag ctgaggaagt gttagagatg caagacttat ggttgttgag gaagccagag
     1501 aaagtgacca gatggttgca gagcaatgga tgggatagac tcaaacgaat ggcggtcagt
     1561 ggagatgact gcgttgtgaa gccaatcgat gataggtttg cacatgccct caggttcttg
     1621 aatgacatgg gaaaagttag gaaagacaca caggagtgga aaccctcgac tggatggagc
     1681 aattgggaag aagtcccgtt ctgctcccac cacttcaaca agctgtacct caaggatggg
     1741 agatccattg tggtcccttg ccgccaccaa gatgaactga ttggccgagc tcgcgtctca
     1801 ccaggggcag gatggagcat ccgggagact gcctgtcttg caaaatcata tgcgcagatg
     1861 tggcagctcc tttatttcca cagaagagac cttcgactga tggctaatgc catttgctcg
     1921 gctgtgccag ttgactgggt accaactggg agaaccacct ggtcaatcca tggaaaggga
     1981 gaatggatga ccactgagga catgctcatg gtgtggaata gagtgtggat tgaggagaac
     2041 gaccatatgg aggacaagac tcctgtaaca aaatggacag acattcccta tctaggaaaa
     2101 agggaggact tatggtgtgg atcccttata gggcacagac cccgcaccac ttgggctgaa
     2161 aacatcaaag acacagtcaa catggtgcgc aggatcatag gtgatgaaga aaagtacatg
     2221 gactatctat ccacccaagt ccgctacttg ggtgaggaag ggtccacacc cggagtgttg
     2281 taaccgcgtc gggtcaccac caccaccacc actgagatcc ggctgctaac aaagcccgaa
     2341 aggaagctga gttggctgct gccaccgctg agcaataact agcataaccc cttggggcct
     2401 ctaaacgggt cttgaggggt tttttgctga aaggaggaac tatatccgga ttggcgaatg
     2461 ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
     2521 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
     2581 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
     2641 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
     2701 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
     2761 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
     2821 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
     2881 taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat
     2941 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3001 aattaattct tagaaaaact catcgagcat caaatgaaac tgcaatttat tcatatcagg
     3061 attatcaata ccatattttt gaaaaagccg tttctgtaat gaaggagaaa actcaccgag
     3121 gcagttccat aggatggcaa gatcctggta tcggtctgcg attccgactc gtccaacatc
     3181 aatacaacct attaatttcc cctcgtcaaa aataaggtta tcaagtgaga aatcaccatg
     3241 agtgacgact gaatccggtg agaatggcaa aagtttatgc atttctttcc agacttgttc
     3301 aacaggccag ccattacgct cgtcatcaaa atcactcgca tcaaccaaac cgttattcat
     3361 tcgtgattgc gcctgagcga gacgaaatac gcgatcgctg ttaaaaggac aattacaaac
     3421 aggaatcgaa tgcaaccggc gcaggaacac tgccagcgca tcaacaatat tttcacctga
     3481 atcaggatat tcttctaata cctggaatgc tgttttcccg gggatcgcag tggtgagtaa
     3541 ccatgcatca tcaggagtac ggataaaatg cttgatggtc ggaagaggca taaattccgt
     3601 cagccagttt agtctgacca tctcatctgt aacatcattg gcaacgctac ctttgccatg
     3661 tttcagaaac aactctggcg catcgggctt cccatacaat cgatagattg tcgcacctga
     3721 ttgcccgaca ttatcgcgag cccatttata cccatataaa tcagcatcca tgttggaatt
     3781 taatcgcggc ctagagcaag acgtttcccg ttgaatatgg ctcataacac cccttgtatt
     3841 actgtttatg taagcagaca gttttattgt tcatgaccaa aatcccttaa cgtgagtttt
     3901 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3961 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4021 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4081 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4141 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4201 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4261 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4321 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4381 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4441 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4501 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4561 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4621 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4681 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc
     4741 ttacgcatct gtgcggtatt tcacaccgca atggtgcact ctcagtacaa tctgctctga
     4801 tgccgcatag ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg
     4861 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     4921 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     4981 tcaccgaaac gcgcgaggca gctgcggtaa agctcatcag cgtggtcgtg aagcgattca
     5041 cagatgtctg cctgttcatc cgcgtccagc tcgttgagtt tctccagaag cgttaatgtc
     5101 tggcttctga taaagcgggc catgttaagg gcggtttttt cctgtttggt cactgatgcc
     5161 tccgtgtaag ggggatttct gttcatgggg gtaatgatac cgatgaaacg agagaggatg
     5221 ctcacgatac gggttactga tgatgaacat gcccggttac tggaacgttg tgagggtaaa
     5281 caactggcgg tatggatgcg gcgggaccag agaaaaatca ctcagggtca atgccagcgc
     5341 ttcgttaata cagatgtagg tgttccacag ggtagccagc agcatcctgc gatgcagatc
     5401 cggaacataa tggtgcaggg cgctgacttc cgcgtttcca gactttacga aacacggaaa
     5461 ccgaagacca ttcatgttgt tgctcaggtc gcagacgttt tgcagcagca gtcgcttcac
     5521 gttcgctcgc gtatcggtga ttcattctgc taaccagtaa ggcaaccccg ccagcctagc
     5581 cgggtcctca acgacaggag cacgatcatg cgcacccgtg gggccgccat gccggcgata
     5641 atggcctgct tctcgccgaa acgtttggtg gcgggaccag tgacgaaggc ttgagcgagg
     5701 gcgtgcaaga ttccgaatac cgcaagcgac aggccgatca tcgtcgcgct ccagcgaaag
     5761 cggtcctcgc cgaaaatgac ccagagcgct gccggcacct gtcctacgag ttgcatgata
     5821 aagaagacag tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg
     5881 actgggttga aggctctcaa gggcatcggt cgagatcccg gtgcctaatg agtgagctaa
     5941 cttacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     6001 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgccagggt
     6061 ggtttttctt ttcaccagtg agacgggcaa cagctgattg cccttcaccg cctggccctg
     6121 agagagttgc agcaagcggt ccacgctggt ttgccccagc aggcgaaaat cctgtttgat
     6181 ggtggttaac ggcgggatat aacatgagct gtcttcggta tcgtcgtatc ccactaccga
     6241 gatatccgca ccaacgcgca gcccggactc ggtaatggcg cgcattgcgc ccagcgccat
     6301 ctgatcgttg gcaaccagca tcgcagtggg aacgatgccc tcattcagca tttgcatggt
     6361 ttgttgaaaa ccggacatgg cactccagtc gccttcccgt tccgctatcg gctgaatttg
     6421 attgcgagtg agatatttat gccagccagc cagacgcaga cgcgccgaga cagaacttaa
     6481 tgggcccgct aacagcgcga tttgctggtg acccaatgcg accagatgct ccacgcccag
     6541 tcgcgtaccg tcttcatggg agaaaataat actgttgatg ggtgtctggt cagagacatc
     6601 aagaaataac gccggaacat tagtgcaggc agcttccaca gcaatggcat cctggtcatc
     6661 cagcggatag ttaatgatca gcccactgac gcgttgcgcg agaagattgt gcaccgccgc
     6721 tttacaggct tcgacgccgc ttcgttctac catcgacacc accacgctgg cacccagttg
     6781 atcggcgcga gatttaatcg ccgcgacaat ttgcgacggc gcgtgcaggg ccagactgga
     6841 ggtggcaacg ccaatcagca acgactgttt gcccgccagt tgttgtgcca cgcggttggg
     6901 aatgtaattc agctccgcca tcgccgcttc cactttttcc cgcgttttcg cagaaacgtg
     6961 gctggcctgg ttcaccacgc gggaaacggt ctgataagag acaccggcat actctgcgac
     7021 atcgtataac gttactggtt tcacattcac caccctgaat tgactctctt ccgggcgcta
     7081 tcatgccata ccgcgaaagg ttttgcgcca ttcgatggtg tccgggatct cgacgctctc
     7141 ccttatgcga ctcctgcatt aggaagcagc ccagtagtag gttgaggccg ttgagcaccg
     7201 ccgccgcaag gaatggtgca tgcaaggaga tggcgcccaa cagtcccccg gccacggggc
     7261 ctgccaccat acccacgccg aaacaagcgc tcatgagccc gaagtggcga gcccgatctt
     7321 ccccatcggt gatgtcggcg atataggcgc cagcaaccgc acctgtggcg ccggtgatgc
     7381 cggccacgat gcgtccggcg tagaggatcg agatctcgat cccgcgaaat
//
