LOCUS       sequence-419615-        6519 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of MERS Mpro.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6519)
  TITLE     The SARS-Cov-2 main protease (Mpro) is responsible for viral 
            polyprotein processing and therefore a therapeutic target.
REFERENCE   2  (bases 1 to 6519)
  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: MERS Mpro - His6-SUMO with autoprocessing -
            MS...
FEATURES             Location/Qualifiers
     source          1..6519
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     150..169
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     253..272
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        253..271
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    272..296
                     /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             311..333
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             344..361
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1636..1653
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1706..1724)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1720..1767
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1804..2259
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1891..1910)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2101..2122
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(2351..3166)
                     /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(2351..3166)
                     /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     3074..3093
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3158..3177
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3288..3876
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3777..3796
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4030..4047
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4062..4201
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4127..4146)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4287..4309
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4303..4494)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(5303..6385)
                     /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     6347..6366
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6386..6463)
                     /gene="lacI"
                     /label=lacI promoter
ORIGIN
        1 aggttgaggc cgttgagcac cgccgccgca aggaatggtg catgcaagga gatggcgccc
       61 aacagtcccc cggccacggg gcctgccacc atacccacgc cgaaacaagc gctcatgagc
      121 ccgaagtggc gagcccgatc ttccccatcg gtgatgtcgg cgatataggc gccagcaacc
      181 gcacctgtgg cgccggtgat gccggccacg atgcgtccgg cgtagaggat cgagatctcg
      241 atcccgcgaa attaatacga ctcactatag gggaattgtg agcggataac aattcccctc
      301 tagaaataat tttgtttaac tttaagaagg agatatacat atgcaccacc atcatcacca
      361 tggctcagga gatcaggaag cgaaaccttc gaccgaggac ctgggcgaca agaaagaagg
      421 ggagtatatt aagttaaagg tgatcggcca ggactcaagc gagatccact tcaaagtcaa
      481 aatgaccacg cacctcaaga agttgaagga gagttactgt caacggcaag gggttcctat
      541 gaactcgctt cgcttcctgt tcgagggcca gcgaattgcg gacaaccaca cacctaagga
      601 acttggtatg gaagaggaag acgtcattga ggtgtaccag gagcagacag gcggcatgtc
      661 cgctgtcatg caatctggtc tcgttaagat gtcccatcct agcggagacg tcgaggcgtg
      721 tatggtacaa gttacgtgcg ggagcatgac gttaaacgga ttgtggctcg acaatacagt
      781 ttggtgccca cgacatgtca tgtgtccggc tgatcaatta agtgacccaa attacgacgc
      841 gttgctcata tccatgacga accacagctt tagcgtccaa aagcacattg gtgcccctgc
      901 taaccttcgg gtcgttggcc atgctatgca gggcactctt ttaaagctga cagttgacgt
      961 tgctaaccct agtacgcctg cctatacatt cactacggtt aaacccggcg ccgcgttctc
     1021 cgttttggca tgttacaacg gtcgtccgac aggtacgttc acagtagtta tgcggccaaa
     1081 ttataccatc aaaggtagct tcctttgcgg ttcatgtggt agtgtgggct acaccaaaga
     1141 gggctcggtg ataaatttct gttacatgca ccaaatggaa cttgcaaacg ggacccacac
     1201 cggtagtgcg tttgatggta ccatgtatgg agctttcatg gacaagcaag ttcaccaagt
     1261 gcaactgacc gataagtact gctctgttaa tgtcgtggct tggctgtatg ccgccatctt
     1321 gaacggctgc gcctggtttg tcaagccaaa tcgcacgtcc gttgtaagct tcaacgagtg
     1381 ggccctggca aaccaattca ctgagttcgt tgggacccag tctgtcgaca tgttggccgt
     1441 taagacgggt gtagcgatag agcagctgct ttacgcaatc caacaattat acacaggttt
     1501 ccagggaaag cagattctcg gttctacaat gttagaggac gaattcacgc ccgaggacgt
     1561 gaacatgcag atcatgggcg ttgtcatgca ataatgactc gagcacccgt cgacaagctt
     1621 gcggccgcac tcgagcacca ccaccaccac cactgagatc cggctgctaa caaagcccga
     1681 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
     1741 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg attggcgaat
     1801 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1861 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1921 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1981 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     2041 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     2101 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     2161 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     2221 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
     2281 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
     2341 gaattaattc ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag
     2401 gattatcaat accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga
     2461 ggcagttcca taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat
     2521 caatacaacc tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat
     2581 gagtgacgac tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt
     2641 caacaggcca gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca
     2701 ttcgtgattg cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa
     2761 caggaatcga atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg
     2821 aatcaggata ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta
     2881 accatgcatc atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg
     2941 tcagccagtt tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat
     3001 gtttcagaaa caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg
     3061 attgcccgac attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat
     3121 ttaatcgcgg cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat
     3181 tactgtttat gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt
     3241 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     3301 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3361 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3421 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3481 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3541 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3601 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3661 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3721 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3781 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3841 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3901 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3961 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     4021 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     4081 cttacgcatc tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg
     4141 atgccgcata gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc
     4201 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4261 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4321 atcaccgaaa cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc
     4381 acagatgtct gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt
     4441 ctggcttctg ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc
     4501 ctccgtgtaa gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat
     4561 gctcacgata cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa
     4621 acaactggcg gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg
     4681 cttcgttaat acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat
     4741 ccggaacata atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa
     4801 accgaagacc attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca
     4861 cgttcgctcg cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag
     4921 ccgggtcctc aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat
     4981 aatggcctgc ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag
     5041 ggcgtgcaag attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa
     5101 gtggtcctcg ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat
     5161 aaagaagaca gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct
     5221 gactgggttg aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta
     5281 acttacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     5341 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg
     5401 tggtttttct tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct
     5461 gagagagttg cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga
     5521 tggtggttaa cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg
     5581 agatatccgc accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca
     5641 tctgatcgtt ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg
     5701 tttgttgaaa accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt
     5761 gattgcgagt gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta
     5821 atgggcccgc taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca
     5881 gtcgcgtacc gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat
     5941 caagaaataa cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat
     6001 ccagcggata gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg
     6061 ctttacaggc ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt
     6121 gatcggcgcg agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg
     6181 aggtggcaac gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg
     6241 gaatgtaatt cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt
     6301 ggctggcctg gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga
     6361 catcgtataa cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct
     6421 atcatgccat accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct
     6481 cccttatgcg actcctgcat taggaagcag cccagtagt
//
