LOCUS       sequence-334215-        6974 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculoviral transfer vector to express SARS-CoV-2 nsp10 in insect 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6974)
  AUTHORS   Canal B, McClure AW, Curran JF, Wu M, Ulferts R, Weissmann F, Zeng 
            J, Bertolin AP, Milligan JC, Basu S, Drury LS, Deegan TD, Fujisawa 
            R, Roberts EL, Basier C, Labib K, Beale R, Howell M, Diffley JFX
  TITLE     Identifying SARS-CoV-2 antiviral compounds by screening for small 
            molecule inhibitors of nsp14/nsp10 exoribonuclease.
  JOURNAL   Biochem J. 2021 Jul 16;478(13):2445-2464. doi: 10.1042/BCJ20210198.
  PUBMED    34198326
REFERENCE   2  (bases 1 to 6974)
  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"; journalName: 
            "Biochem J"; date: "2021-07-16- 16"; volume: "478"; issue: "13"; 
            pages: "2445-2464"
COMMENT     Sequence Label: pBIG1b_nsp10-6His-3xFlag (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6974
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(356..361,362..378,379..384))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(780..813)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(859..878)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    903..951
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        1046..1137
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     1062..1079
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      1181..1190
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1616..1633
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1640..1705
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1838..1972
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1875..1894)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1929..1948
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  2287..2452
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     CDS             2976..3767
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3808..4262
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3895..3914)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4104..4125
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4288..4392
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4393..4461,4462..5253)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4611..4630)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5424..6012
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5913..5932
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6166..6183
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6317..6541
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(join(6608..6974,1..167))
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 gtggctacgt ctccgaactc acgaccgaaa agatcaagag cagcccgcat ggatttgact
       61 tggtcagggc cgagcctaca tgtgcgaatg atgcccatac ttgagccacc taactttgtt
      121 ttagggcgac tgccctgctg cgtaacatcg ttgctgctgc gtaacatcgt tgctgctcca
      181 taacatcaaa catcgaccca cggcgtaacg cgcttgctgc ttggatgccc gaggcataga
      241 ctgtacaaaa aaacagtcat aacaagccat gaaaaccgcc actgcgccgt taccaccgct
      301 gcgttcggtc aaggttctgg accagttgcg tgagcgcata cgctacttgc attacagttt
      361 acgaaccgaa caggcttatg tcaactgggt ttgtgccttc atccgtttcc acggtgtgcg
      421 tcacccggca accttgggca gcagcgaagt cgaggcattt ctgtcctggc tggcgaacga
      481 gcgcaaggtt tcggtctcca cgcatcgtca ggcattggcg gccttgctgt tcttctacgg
      541 caaggtgctg tgcacggatc tgcccttgct tcaggagatc ggtagacctc ggccgtcgcg
      601 gcgcttgccg gtggtgctga ccccggatga agtggttcgc atcctcggtt ttctggaagg
      661 cgagcatcgt ttgttcgccc aggactctag ctatagttct agtggttggc tacagctttg
      721 tttgtactat caacaggttg aactgctgat caacagatcc tctacgcggc cgcggtacca
      781 taacttcgta tagcatacat tatacgaagt tatctgggtt taaactatat ctcaatcggg
      841 gtttttatcg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
      901 gacggcaata aaaagacaga ataaaacgca cgggtgttgg gtcgtttgtt cataaacgcg
      961 gggttcggtc ccagggctgg cactctgtca agtgaacgct ctatggtcta aagatttaaa
     1021 tcgacctact ccggaatatt aatagatcat ggagataatt aaaatgataa ccatctcgca
     1081 aataaataag tattttactg ttttcgtaac agttttgtaa taaaaaaacc tataaatatt
     1141 ccggattatt cataccgtcc caccatcggg cgcggatcca gccaccatgg ctggaaacgc
     1201 taccgaggtg cccgctaact ctaccgtgct gtccttctgc gctttcgctg tggacgctgc
     1261 taaggcttac aaggactacc tggcttccgg tggccagcct atcaccaact gcgtgaagat
     1321 gctgtgcacc cacaccggta ctggacaggc tatcactgtg acccctgagg ctaacatgga
     1381 ccaagagtcc ttcggtggtg cttcctgctg cctgtactgc cgttgccaca tcgatcaccc
     1441 caatcctaag ggcttctgcg acctgaaggg caaatacgtg cagatcccca ccacttgcgc
     1501 taacgaccct gtgggtttca ccctgaagaa caccgtgtgc accgtctgcg gcatgtggaa
     1561 aggatacggt tgctcctgcg accagctgcg cgaacccatg ttgcagggtg gctcccacca
     1621 tcaccaccat cacggttctg actacaagga ccacgacggc gattacaagg atcatgacat
     1681 cgactacaaa gacgacgatg acaagtaaaa gcttgtcgag aagtactaga ggatcataat
     1741 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1801 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1861 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1921 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tctgatcact
     1981 gcttgagcct agaagatccg gctgctaaca aagcccgaaa ggaagctgag ttggctgctg
     2041 ccaccgctga gcaataacta tcataaccat ttaaataaat ctatatctca atcggggtta
     2101 cgtcgatacc ccaccgagac cccattggga ccaatacgcc cgcgtttctt ccttttcccc
     2161 accccaaccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcaag
     2221 ccctgccata gccactacgg gttaattaaa aactggatac tattgcacgt ttaaacagga
     2281 gatccgaacc agataagtga aatctagttc caaactattt tgtcattttt aattttcgta
     2341 ttagcttacg acgctacacc cagttcccat ctattttgtc actcttccct aaataatcct
     2401 taaaaactcc atttccaccc ctcccagttc ccaactattt tgtccgccca cagcggggca
     2461 tttttcttcc tgttatgttt ttaatcaaac atcctgccaa ctccatgtga caaaccgtca
     2521 tcttcggcta ctttttctct gtcacagaat gaaaattttt ctgtcatctc ttcgttatta
     2581 atgtttgtaa ttgactgaat atcaacgctt atttgcagcc tgaactggga ggcagaataa
     2641 atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca
     2701 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaag tagcgtatgc
     2761 gctcacgcaa ctggtccaga accttgaccg aacgcagcgg tggtaacggc gcagtggcgg
     2821 ttttcatggc ttgttatgac tgtttttttg gggtacagtc tatgcctcgg gcatccaagc
     2881 agcaagcgcg ttacgccgtg ggtcgatgtt tgatgttatg gagcagcaac gatgttacgc
     2941 agcagggcag tcgccctaaa acaaagttaa acatcatgag ggaagcggtg atcgccgaag
     3001 tatcgactca actatcagag gtagttggcg tcatcgagcg ccatctcgaa ccgacgttgc
     3061 tggccgtaca tttgtacggc tccgcagtgg atggcggcct gaagccacac agtgatattg
     3121 atttgctggt tacggtgacc gtaaggcttg atgaaacaac gcggcgagct ttgatcaacg
     3181 accttttgga aacttcggct tcccctggag agagcgagat tctccgcgct gtagaagtca
     3241 ccattgttgt gcacgacgac atcattccgt ggcgttatcc agctaagcgc gaactgcaat
     3301 ttggagaatg gcagcgcaat gacattcttg caggtatctt cgagccagcc acgatcgaca
     3361 ttgatctggc tatcttgctg acaaaagcaa gagaacatag cgttgccttg gtaggtccag
     3421 cggcggagga actctttgat ccggttcctg aacaggatct atttgaggcg ctaaatgaaa
     3481 ccttaacgct atggaactcg ccgcccgact gggctggcga tgagcgaaat gtagtgctta
     3541 cgttgtcccg catttggtac agcgcagtaa ccggcaaaat cgcgccgaag gatgtcgctg
     3601 ccgactgggc aatggagcgc ctgccggccc agtatcagcc cgtcatactt gaagctagac
     3661 aggcttatct tggacaagaa gaagatcgct tggcctcgcg cgcagatcag ttggaagaat
     3721 ttgtccacta cgtgaaaggc gagatcacca aggtagtcgg caaataatgt ctaacaattc
     3781 gttcaagccg acggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     3841 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3901 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3961 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     4021 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     4081 cgttggagtc cacgttctta atagtggact cttgttccaa actggaacaa cactcaaccc
     4141 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     4201 aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat
     4261 ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4321 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4381 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4441 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4501 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4561 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4621 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4681 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     4741 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     4801 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     4861 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     4921 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     4981 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5041 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5101 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5161 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5221 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5281 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5341 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5401 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5461 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5521 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactat tcttctagtg
     5581 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5641 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5701 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5761 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5821 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5881 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5941 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6001 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6061 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6121 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6181 gaggaagcgg aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     6241 caccgcatag accagccgcg taacctggca aaatcggtta cggttgagta ataaatggat
     6301 gccctgcgta agcgggtgtg ggcggacaat aaagtcttaa actgaacaaa atagatctaa
     6361 actatgacaa taaagtctta aactagacag aatagttgta aactgaaatc agtccagtta
     6421 tgctgtgaaa aagcatactg gacttttgtt atggctaaag caaactcttc attttctgaa
     6481 gtgcaaattg cccgtcgtat taaagagggg cgtggccaag ggcatggtaa agactatatt
     6541 cgcggcgttg tgacaattta ccgaacaact ccgcggccgg gaagccgatc tcggcttgaa
     6601 cgaattgtta ggtggcggta cttgggtcga tatcaaagtg catcacttct tcccgtatgc
     6661 ccaactttgt atagagagcc actgcgggat cgtcaccgta atctgcttgc acgtagatca
     6721 cataagcacc aagcgcgttg gcctcatgct tgaggagatt gatgagcgcg gtggcaatgc
     6781 cctgcctccg gtgctcgccg gagactgcga gatcatagat atagatctca ctacgcggct
     6841 gctcaaactt gggcagaacg taagccgcga gagcgccaac aaccgcttct tggtcgaagg
     6901 cagcaagcgc gatgaatgtc ttactacgga gcaagttccc gaggtaatcg gagtccggct
     6961 gatgttggga gtag
//
