LOCUS       sequence-329229-        5798 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of the SARS-CoV-2 protein nsp10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5798)
  AUTHORS   Perveen S, Khalili Yazdi A, Devkota K, Li F, Ghiabi P, Hajian T, 
            Loppnau P, Bolotokova A, Vedadi M
  TITLE     A High-Throughput RNA Displacement Assay for Screening SARS-CoV-2 
            nsp10-nsp16 Complex toward Developing Therapeutics for COVID-19.
  JOURNAL   SLAS Discov. 2021 Jan 10:2472555220985040. doi: 
            10.1177/2472555220985040.
  PUBMED    33423577
REFERENCE   2  (bases 1 to 5798)
  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: "SLAS 
            Discov. 2021 Jan 10:2472555220985040. doi: 
            10.1177/2472555220985040."
COMMENT     Sequence Label: p28BIOHTEV-LIC: nsp10_SARS-CoV-2
FEATURES             Location/Qualifiers
     source          1..5798
                     /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..138
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             160..180
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             616..633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             663..680
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(733..751)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      747..794
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      831..1286
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(918..937)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1128..1149
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1378..2193)
                     /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(1378..2193)
                     /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     2101..2120
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2185..2204
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2315..2903
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2804..2823
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3057..3074
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3089..3228
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3154..3173)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3314..3336
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3330..3521)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4330..5412)
                     /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     5374..5393
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5413..5490)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5696..5715
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg agcggcctga acgatatttt tgaagcgcag
      121 aaaattgaat ggcatgaagg cagcgctgga ggttcaggtg aaaacttgta tttccagggc
      181 gccggtaatg caaccgaagt gcccgctaac agcactgttc tttccttctg cgccttcgcg
      241 gtggatgcgg ctaaagccta taaagactat cttgcttctg ggggtcaacc tattactaat
      301 tgtgtcaaaa tgctttgcac ccacacaggg actggacagg ctattaccgt gacccctgag
      361 gctaatatgg atcaggaatc ctttggaggc gcgtcctgtt gcctgtactg ccgttgtcat
      421 attgaccatc ccaatcctaa agggttttgc gacttaaaag gaaagtatgt tcaaattcca
      481 acgacttgcg caaatgaccc cgtgggattc acattgaaaa acactgtatg cactgtctgc
      541 gggatgtgga aaggatacgg gtgttcatgc gatcaattac gcgagcctat gcttcagtga
      601 tgaggtggaa gtggtcatca ccaccatcat cactgatgac gaagcttgcg gccgcactcg
      661 agcaccacca ccaccaccac tgagatccgg ctgctaacaa agcccgaaag gaagctgagt
      721 tggctgctgc caccgctgag caataactag cataacccct tggggcctct aaacgggtct
      781 tgaggggttt tttgctgaaa ggaggaacta tatccggatt ggcgaatggg acgcgccctg
      841 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
      901 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
      961 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     1021 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     1081 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     1141 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     1201 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt
     1261 taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc
     1321 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgaa ttaattctta
     1381 gaaaaactca tcgagcatca aatgaaactg caatttattc atatcaggat tatcaatacc
     1441 atatttttga aaaagccgtt tctgtaatga aggagaaaac tcaccgaggc agttccatag
     1501 gatggcaaga tcctggtatc ggtctgcgat tccgactcgt ccaacatcaa tacaacctat
     1561 taatttcccc tcgtcaaaaa taaggttatc aagtgagaaa tcaccatgag tgacgactga
     1621 atccggtgag aatggcaaaa gtttatgcat ttctttccag acttgttcaa caggccagcc
     1681 attacgctcg tcatcaaaat cactcgcatc aaccaaaccg ttattcattc gtgattgcgc
     1741 ctgagcgaga cgaaatacgc gatcgctgtt aaaaggacaa ttacaaacag gaatcgaatg
     1801 caaccggcgc aggaacactg ccagcgcatc aacaatattt tcacctgaat caggatattc
     1861 ttctaatacc tggaatgctg ttttcccggg gatcgcagtg gtgagtaacc atgcatcatc
     1921 aggagtacgg ataaaatgct tgatggtcgg aagaggcata aattccgtca gccagtttag
     1981 tctgaccatc tcatctgtaa catcattggc aacgctacct ttgccatgtt tcagaaacaa
     2041 ctctggcgca tcgggcttcc catacaatcg atagattgtc gcacctgatt gcccgacatt
     2101 atcgcgagcc catttatacc catataaatc agcatccatg ttggaattta atcgcggcct
     2161 agagcaagac gtttcccgtt gaatatggct cataacaccc cttgtattac tgtttatgta
     2221 agcagacagt tttattgttc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     2281 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     2341 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     2401 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     2461 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     2521 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     2581 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     2641 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     2701 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     2761 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     2821 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     2881 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     2941 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     3001 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     3061 agtcagtgag cgaggaagcg gaagagcgcc tgatgcggta ttttctcctt acgcatctgt
     3121 gcggtatttc acaccgcaat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     3181 aagccagtat acactccgct atcgctacgt gactgggtca tggctgcgcc ccgacacccg
     3241 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     3301 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     3361 gcgaggcagc tgcggtaaag ctcatcagcg tggtcgtgaa gcgattcaca gatgtctgcc
     3421 tgttcatccg cgtccagctc gttgagtttc tccagaagcg ttaatgtctg gcttctgata
     3481 aagcgggcca tgttaagggc ggttttttcc tgtttggtca ctgatgcctc cgtgtaaggg
     3541 ggatttctgt tcatgggggt aatgataccg atgaaacgag agaggatgct cacgatacgg
     3601 gttactgatg atgaacatgc ccggttactg gaacgttgtg agggtaaaca actggcggta
     3661 tggatgcggc gggaccagag aaaaatcact cagggtcaat gccagcgctt cgttaataca
     3721 gatgtaggtg ttccacaggg tagccagcag catcctgcga tgcagatccg gaacataatg
     3781 gtgcagggcg ctgacttccg cgtttccaga ctttacgaaa cacggaaacc gaagaccatt
     3841 catgttgttg ctcaggtcgc agacgttttg cagcagcagt cgcttcacgt tcgctcgcgt
     3901 atcggtgatt cattctgcta accagtaagg caaccccgcc agcctagccg ggtcctcaac
     3961 gacaggagca cgatcatgcg cacccgtggg gccgccatgc cggcgataat ggcctgcttc
     4021 tcgccgaaac gtttggtggc gggaccagtg acgaaggctt gagcgagggc gtgcaagatt
     4081 ccgaataccg caagcgacag gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg
     4141 aaaatgaccc agagcgctgc cggcacctgt cctacgagtt gcatgataaa gaagacagtc
     4201 ataagtgcgg cgacgatagt catgccccgc gcccaccgga aggagctgac tgggttgaag
     4261 gctctcaagg gcatcggtcg agatcccggt gcctaatgag tgagctaact tacattaatt
     4321 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     4381 atcggccaac gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt
     4441 caccagtgag acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag
     4501 caagcggtcc acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttaacgg
     4561 cgggatataa catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc
     4621 aacgcgcagc ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc
     4681 aaccagcatc gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc
     4741 ggacatggca ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag
     4801 atatttatgc cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa
     4861 cagcgcgatt tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc
     4921 ttcatgggag aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc
     4981 cggaacatta gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt
     5041 aatgatcagc ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc
     5101 gacgccgctt cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga
     5161 tttaatcgcc gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc
     5221 aatcagcaac gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag
     5281 ctccgccatc gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt
     5341 caccacgcgg gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt
     5401 tactggtttc acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc
     5461 gcgaaaggtt ttgcgccatt cgatggtgtc cgggatctcg acgctctccc ttatgcgact
     5521 cctgcattag gaagcagccc agtagtaggt tgaggccgtt gagcaccgcc gccgcaagga
     5581 atggtgcatg caaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
     5641 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
     5701 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
     5761 gtccggcgta gaggatcgag atctcgatcc cgcgaaat
//
