LOCUS       sequence-473707-        5721 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial protein expression of NSP10.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5721)
  AUTHORS   Rona G, Zeke A, Miwatani-Minter B, de Vries M, Kaur R, Schinlever A,
            Garcia SF, Goldberg HV, Wang H, Hinds TR, Bailly F, Zheng N, Cotelle
            P, Desmaele D, Landau NR, Dittmann M, Pagano M
  TITLE     The NSP14/NSP10 RNA repair complex as a Pan-coronavirus therapeutic 
            target.
  JOURNAL   Cell Death Differ. 2022 Feb;29(2):285-292. doi: 
            10.1038/s41418-021-00900-1. Epub 2021 Dec 3.
  PUBMED    34862481
REFERENCE   2  (bases 1 to 5721)
  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: 
            "10.1038/s41418-021-00900-1"; journalName: "Cell Death Differ"; 
            date: "2022-02"; volume: "29"; issue: "2"; pages: "285-292"
COMMENT     Sequence Label: pET-30a-NSP10-SARS-CoV
FEATURES             Location/Qualifiers
     source          1..5721
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1576..1767
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1761..1783)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1869..2011
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1924..1943
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2026..2043)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2197..2785)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2277..2296)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(2896..2915)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2907..3722
                     /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             2907..3722
                     /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     complement(2980..2999)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(3815..4270)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3952..3973)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4164..4183
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4307..4354)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4350..4368
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4421..4438)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4466..4483)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4935..4957
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4972..4996
                     /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     complement(4996..5015)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4997..5015)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5103..5122)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5328..5405
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5406..5721,1..767)
                     /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(5425..5444)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg atcattaact
       61 atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat gttccggcgt
      121 tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc catgaagacg
      181 gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc gcgctgttag
      241 cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat aaatatctca
      301 ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc atgtccggtt
      361 ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg ctggttgcca
      421 acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg cgcgttggtg
      481 cggacatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat atcccgccgt
      541 taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac cgcttgctgc
      601 aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca ctggtgaaaa
      661 gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
      721 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
      781 aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt gagagccttc
      841 aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact
      901 gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt cattttcggc
      961 gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt attcggaatc
     1021 ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag
     1081 caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg
     1141 aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc accgatacgc
     1201 gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg
     1261 gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt
     1321 atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc tacatctgta
     1381 ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg catccatacc
     1441 gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc agtaacccgt
     1501 atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag aaatccccct
     1561 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
     1621 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga tgaacaggca
     1681 gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg cctcgcgcgt
     1741 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     1801 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     1861 tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac tggcttaact
     1921 atgcggcatc agagcagatt gtactgagag tgcaccatat atgcggtgtg aaataccgca
     1981 cagatgcgta aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc
     2041 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     2101 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     2161 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     2221 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     2281 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     2341 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     2401 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     2461 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     2521 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     2581 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     2641 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     2701 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     2761 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     2821 tcagtggaac gaaaactcac gttaagggat tttggtcatg aacaataaaa ctgtctgctt
     2881 acataaacag taatacaagg ggtgttatga gccatattca acgggaaacg tcttgctcta
     2941 ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg gctcgcgata
     3001 atgtcgggca atcaggtgcg acaatctatc gattgtatgg gaagcccgat gcgccagagt
     3061 tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag atggtcagac
     3121 taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc cgtactcctg
     3181 atgatgcatg gttactcacc actgcgatcc ccgggaaaac agcattccag gtattagaag
     3241 aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg cgccggttgc
     3301 attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg
     3361 cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac gagcgtaatg
     3421 gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc tcaccggatt
     3481 cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag gggaaattaa
     3541 taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat cttgccatcc
     3601 tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt caaaaatatg
     3661 gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat gagtttttct
     3721 aagaattaat tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     3781 gttccgcgca catttccccg aaaagtgcca cctgaaattg taaacgttaa tattttgtta
     3841 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     3901 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     3961 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4021 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     4081 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     4141 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     4201 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     4261 cagggcgcgt cccattcgcc aatccggata tagttcctcc tttcagcaaa aaacccctca
     4321 agacccgttt agaggcccca aggggttatg ctagttattg ctcagcggtg gcagcagcca
     4381 actcagcttc ctttcgggct ttgttagcag ccggatctca gtggtggtgg tggtggtgct
     4441 cgagtgcggc cgcaagcttt cattagtggt ggtggtggtg atggctgcca ccaccggaac
     4501 cgccaccctg catcagcggc tcacgcagct ggtcgcagct acaaccataa cctttccaca
     4561 taccgcacac ggtgcatacg gtgttgcgca aggtgaaacc aaccgggtcg ttagcgcacg
     4621 tggtcggaat ctgcacgtat ttacccttta agtcgcaaaa gcctttcgga ttcggatggt
     4681 cgatatgaca acggcagtac aggcaacaag atgcgccgcc gaacgactct tgatccatgt
     4741 tggcttccgg ggtaaccgtg atcgcttggc ctgtaccagt atgggtacac agcatcttca
     4801 cgcaattggt aatcggttgg ccgccggaag ccaagtaatc tttatacgcc ttcgctggat
     4861 caactgcaaa cgcgcagaag ctcaggacgg tgctgttagc gggtacttct gtcgcatttc
     4921 ctgccatatg tatatctcct tcttaaagtt aaacaaaatt atttctagag gggaattgtt
     4981 atccgctcac aattccccta tagtgagtcg tattaatttc gcgggatcga gatcgatctc
     5041 gatcctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     5101 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     5161 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     5221 ctccttgcat gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     5281 ctgcttccta atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat
     5341 ggcgcaaaac ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg
     5401 tgaatgtgaa accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga
     5461 ccgtttcccg cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg
     5521 aagcggcgat ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca
     5581 aacagtcgtt gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa
     5641 ttgtcgcggc gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg
     5701 tagaacgaag cggcgtcgaa g
//
