LOCUS       sequence-334004-        6707 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculoviral transfer vector to express untagged nsp7 (SARS-CoV-2) in
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6707)
  AUTHORS   Bertolin AP, Weissmann F, Zeng J, Posse V, Milligan JC, Canal B, 
            Ulferts R, Wu M, Drury LS, Howell M, Beale R, Diffley JFX
  TITLE     Identifying SARS-CoV-2 antiviral compounds by screening for small 
            molecule inhibitors of nsp12/7/8 RNA-dependent RNA polymerase.
  JOURNAL   Biochem J. 2021 Jul 16;478(13):2425-2443. doi: 10.1042/BCJ20210200.
  PUBMED    34198323
REFERENCE   2  (bases 1 to 6707)
  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: "2425-2443"
COMMENT     Sequence Label: pBIG1b_nsp7 (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6707
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     mobile_element  636..801
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     polyA_signal    1116..1250
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1140..1159)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1194..1213
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1684..1775)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(1742..1759)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    1870..1918
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     1943..1962
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     protein_bind    2008..2041
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(2437..2442,2443..2459,2460..2465)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             2654..3187
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  3254..3478
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(3612..3629)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3783..4371)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3863..3882)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4542..5333,5334..5402))
                     /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     5165..5184
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5403..5507)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5533..5987)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5670..5691)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5881..5900
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(6028..6707,1..112))
                     /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"
ORIGIN
        1 ctgcggagcc gtacaaatgt acggccagca acgtcggttc gagatggcgc tcgatgacgc
       61 caactacctc tgatagttga gtcgatactt cggcgatcac cgcttccctc atgatgttta
      121 actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat aacatcaaac
      181 atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtaccccaa
      241 aaaaacagtc ataacaagcc atgaaaaccg ccactgcgcc gttaccaccg ctgcgttcgg
      301 tcaaggttct ggaccagttg cgtgagcgca tacgctactt accggtttat tgactaccgg
      361 aagcagtgtg accgtgtgct tctcaaatgc ctgaggccag tttgctcagg ctctccccgt
      421 ggaggtaata attgacgata tgatcattta ttctgcctcc cagttcaggc tgcaaataag
      481 cgttgatatt cagtcaatta caaacattaa taacgaagag atgacagaaa aattttcatt
      541 ctgtgacaga gaaaaagtag ccgaagatga cggtttgtca catggagttg gcaggatgtt
      601 tgattaaaaa cataacagga agaaaaatgc cccgctgtgg gcggacaaaa tagttgggaa
      661 ctgggagggg tggaaatgga gtttttaagg attatttagg gaagagtgac aaaatagatg
      721 ggaactgggt gtagcgtcgt aagctaatac gaaaattaaa aatgacaaaa tagtttggaa
      781 ctagatttca cttatctggt tcggatctcc tgtttaaacg tgcaatagta tccagttttt
      841 aattaacccg tagtggctat ggcagggctt gccgccccga cgttggctgc gagccctggg
      901 ccttcacccg aacttggggg ttggggtggg gaaaaggaag aaacgcgggc gtattggtcc
      961 caatggggtc tcggtggggt atcgacgtaa ccccgattga gatatagatt tatttaaatg
     1021 gttatgatag ttattgctca gcggtggcag cagccaactc agcttccttt cgggctttgt
     1081 tagcagccgg atcttctagg ctcaagcagt gatcagatcc agacatgata agatacattg
     1141 atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt
     1201 gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca
     1261 attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt taaagcaagt
     1321 aaaacctcta caaatgtggt atggctgatt atgatcctct agtacttctc gacaagcttt
     1381 tactgcaggg tagcacggtt gtccagcatc tcttcgcaca gcttgttgat gtccacagca
     1441 ccttgcatag acagcaggac ggacagcagg gacaccatct tctcgaaagc ctcggtggtg
     1501 tccttagcca gcaggatgtc gttgtgcagt tggacgcact gagcccacag cttgctggag
     1561 gattccacac gcagctgttg cagcacagac agcagcacca cggaggtgca cttcacgtcg
     1621 gacatcttgg acatggtggc tggatccgcg cccgatggtg ggacggtatg aataatccgg
     1681 aatatttata ggttttttta ttacaaaact gttacgaaaa cagtaaaata cttatttatt
     1741 tgcgagatgg ttatcatttt aattatctcc atgatctatt aatattccgg agtaggtcga
     1801 tttaaatctt tagaccatag agcgttcact tgacagagtg ccagccctgg gaccgaaccc
     1861 cgcgtttatg aacaaacgac ccaacacccg tgcgttttat tctgtctttt tattgccgtc
     1921 atagcgcggg ttccttccgg tattgtctcc ttccgtgttt cagttagcct cgataaaaac
     1981 cccgattgag atatagttta aacccagata acttcgtata atgtatgcta tacgaagtta
     2041 tggtaccgcg gccgcgtaga ggatctgttg atcagcagtt caacctgttg atagtacaaa
     2101 caaagctgta gccaaccact agaactatag ctagagtcct gggcgaacaa acgatgctcg
     2161 ccttccagaa aaccgaggat gcgaaccact tcatccgggg tcagcaccac cggcaagcgc
     2221 cgcgacggcc gaggtctacc gatctcctga agcaagggca gatccgtgca cagcaccttg
     2281 ccgtagaaga acagcaaggc cgccaatgcc tgacgatgcg tggagaccga aaccttgcgc
     2341 tcgttcgcca gccaggacag aaatgcctcg acttcgctgc tgcccaaggt tgccgggtga
     2401 cgcacaccgt ggaaacggat gaaggcacaa acccagttga cataagcctg ttcggttcgt
     2461 aaactgtaat gcaagtagcg tatgcgctca cgcaactggt ccagaacctt gaccgaacgc
     2521 agcggtggta acggcgcagt ggcggttttc atggcttgtt atgactgttt ttttgtacag
     2581 tctatgcctc gggcatccaa gcagcaagcg cgttacgccg tgggtcgatg tttgatgtta
     2641 tggagcagca acgatgttac gcagcagcaa cgatgttacg cagcagggca gtcgccctaa
     2701 aacaaagtta ggtggctcaa gtatgggcat cattcgcaca tgtaggctcg gccctgacca
     2761 agtcaaatcc atgcgggctg ctcttgatct tttcggtcgt gagttcggag acgtagccac
     2821 ctactcccaa catcagccgg actccgatta cctcgggaac ttgctccgta gtaagacatt
     2881 catcgcgctt gctgccttcg accaagaagc ggttgttggc gctctcgcgg cttacgttct
     2941 gcccaagttt gagcagccgc gtagtgagat ctatatctat gatctcgcag tctccggcga
     3001 gcaccggagg cagggcattg ccaccgcgct catcaatctc ctcaagcatg aggccaacgc
     3061 gcttggtgct tatgtgatct acgtgcaagc agattacggt gacgatcccg cagtggctct
     3121 ctatacaaag ttgggcatac gggaagaagt gatgcacttt gatatcgacc caagtaccgc
     3181 cacctaacaa ttcgttcaag ccgagatcgg cttcccggcc gcggagttgt tcggtaaatt
     3241 gtcacaacgc cgcgaatata gtctttacca tgcccttggc cacgcccctc tttaatacga
     3301 cgggcaattt gcacttcaga aaatgaagag tttgctttag ccataacaaa agtccagtat
     3361 gctttttcac agcataactg gactgatttc agtttacaac tattctgtct agtttaagac
     3421 tttattgtca tagtttagat ctattttgtt cagtttaaga ctttattgtc cgcccacacc
     3481 cgcttacgca gggcatccat ttattactca accgtaaccg attttgccag gttacgcggc
     3541 tggtctatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc
     3601 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     3661 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     3721 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     3781 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     3841 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     3901 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     3961 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4021 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4081 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4141 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4201 cctaactacg gctacactag aagaatagta tttggtatct gcgctctgct gaagccagtt
     4261 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4321 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     4381 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     4441 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     4501 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     4561 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     4621 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     4681 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     4741 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     4801 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     4861 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     4921 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     4981 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5041 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5101 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5161 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     5221 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     5281 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     5341 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     5401 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     5461 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     5521 aaagtgccac ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt
     5581 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
     5641 aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaagaa
     5701 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     5761 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     5821 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     5881 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     5941 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccaat
     6001 ccgtcggctt gaacgaattg ttagacatta tttgccgact accttggtga tctcgccttt
     6061 cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat cttcttcttg
     6121 tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg gcaggcgctc
     6181 cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg cgctgtacca
     6241 aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg gcggcgagtt
     6301 ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa ccggatcaaa
     6361 gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt ttgtcagcaa
     6421 gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa tgtcattgcg
     6481 ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa tgatgtcgtc
     6541 gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag gggaagccga
     6601 agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc ttacggtcac
     6661 cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatcca
//
