LOCUS       sequence-308774-        3666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SARS-CoV-2 nsp8.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3666)
  AUTHORS   Hillen HS, Kokic G, Farnung L, Dienemann C, Tegunov D, Cramer P
  TITLE     Structure of replicating SARS-CoV-2 polymerase.
  JOURNAL   Nature. 2020 May 21. pii: 10.1038/s41586-020-2368-8. doi: 
            10.1038/s41586-020-2368-8.
  PUBMED    32438371
REFERENCE   2  (bases 1 to 3666)
  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: "Nature. 
            2020 May 21. pii: 10.1038/s41586-020-2368-8. doi: 
            10.1038/s41586-020-2368-8."
COMMENT     Sequence Label: pET-His-SCov2-nsp8
FEATURES             Location/Qualifiers
     source          1..3666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             36..58
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             77..94
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             95..115
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     terminator      761..807
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      1974..2013
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     rep_origin      2044..2632
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2533..2552
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2786..2803
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2818..2957
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2883..2902)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3043..3065
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3059..3250)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      3508..3554
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3614..3641
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
ORIGIN
        1 taatacgact cactataggc ccctctagaa ataattttgt ttaactttaa gaaggagata
       61 taccatgggt tcttctcacc atcaccatca ccatgaaaac ctgtacttcc aatccaatgc
      121 agcaattgca agcgaattta gcagcctgcc gagctatgca gcatttgcaa ccgcacaaga
      181 ggcatatgaa caggcagttg ccaatggtga tagcgaagtt gttctgaaaa aactgaaaaa
      241 gtcactgaac gtggccaaat ccgaatttga tcgtgatgca gcaatgcagc gtaaactgga
      301 aaaaatggca gatcaggcaa tgacccagat gtataaacag gcacgtagcg aagataaacg
      361 tgcaaaagtt accagcgcaa tgcagaccat gctgtttacc atgctgcgca aactggataa
      421 tgatgccctg aacaacatta tcaataatgc ccgtgatggt tgtgttccgc tgaacattat
      481 tccgctgacc accgcagcaa aactgatggt ggttattccg gattacaaca cctataaaaa
      541 cacctgtgat ggcaccacct ttacctatgc aagcgcactg tgggaaattc agcaggttgt
      601 tgatgcagat agcaaaattg ttcagctgag cgaaattagc atggataata gcccgaatct
      661 ggcatggcct ctgattgtta ccgcactgcg tgcaaatagc gcagttaaac tgcagtaata
      721 acattggaag tggataactg gctgctgcca ccgctgagca tagcataacc ccttggggcc
      781 tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatcaag tcccgaagta
      841 gctgtcagac tgctggcgac cgccgcaact agtggcgtca tgcgtgccgt atggtctttc
      901 gcctgtagac gccctacaag ccgtactaaa gtggaaagac ccgcaaaagg tatcgcgatc
      961 gccctgcagg gaactatatc cgactagtag cccgcctaat gagcgggctt ttttttaatt
     1021 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     1081 cctgataaat gcttcgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     1141 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctcgtgaaag
     1201 taaaagacgc agaggaccaa ttgggggcac gagtgggata catagaactg gacttgaata
     1261 gcggtaaaat ccttgagagt tttcgccctg aagagcgttt tccaatgatg agcactttca
     1321 aagttctgct atgtggagca gtattatccc gtgtagatgc ggggcaagag caactcggac
     1381 gacgaataca ctattcgcag aatgacttgg ttgaatactc cccagtgaca gaaaagcacc
     1441 ttacggacgg aatgacggta agagaattat gtagtgccgc cataacgatg agtgataaca
     1501 ctgcggcgaa cttacttctg acaaccatcg gtggaccgaa ggaattaacc gcttttttgc
     1561 acaatatggg agaccatgta actcgccttg accgttggga accagaactg aatgaagcca
     1621 taccaaacga cgagcgagac accacaatgc ctgcggcaat ggcaacaaca ttacgcaaac
     1681 tattaactgg cgaactactt actctggctt cacggcaaca attaatagac tggcttgaag
     1741 cggataaagt tgcaggacca ctactgcgtt cggcacttcc tgctggctgg tttattgctg
     1801 ataaatctgg ggcaggagag cgtggttcac ggggtatcat tgccgcactt ggaccagatg
     1861 gtaagccttc ccgtatcgta gttatctaca cgacgggtag tcaggcaact atggacgaac
     1921 gaaatagaca gattgctgaa ataggggctt cactgattaa gcattggtaa accgatacaa
     1981 ttaaaggctc cttttggagc cttttttttt ggacggaccg gtagaaaaga tcaaaggatc
     2041 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2101 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2161 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2221 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2281 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2341 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2401 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2461 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2521 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2581 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2641 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     2701 tgcgttatcc cctgattctg tggataaccg tattaccgcc cttgagtgag ctgataccgc
     2761 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     2821 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcaatg gtgcactctc
     2881 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     2941 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3001 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3061 agaggttttc accgtcatca ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt
     3121 ggtcgtgaag cgattcacag atgtctgcct gttcatccgc gtccagctcg ttgagtttct
     3181 ccagaagcgt taatgtctgg cttctgataa agcgggccat gttaagggcg gttttttcct
     3241 gtttggtcac tgatgcctcc gtgtaagggg gatttctgtt catggggtaa tgataccgat
     3301 gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga
     3361 acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca
     3421 gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ggcaccatgc atcgatcccg
     3481 cgaaatgcgg ccgcttaatt aaggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     3541 gcctttcgtt ttatctgttg tttgtcggtg agtccttcca gtctgggcga aagcgcggtc
     3601 gatcaaatta agcagaaggc catcctgacg gatggccttt ttgcgtttct atcgatcccg
     3661 cgaaat
//
