LOCUS       sequence-334012-        7319 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculoviral transfer vector to express untagged nsp7-Linker-nsp8 
            fusion protein (SARS-CoV-2) in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7319)
  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 7319)
  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-Linker-nsp8 (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..7319
                     /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(2296..2387)
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(2354..2371)
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     polyA_signal    2482..2530
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2555..2574
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     protein_bind    2620..2653
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        join(3049..3054,3055..3071,3072..3077)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             3266..3799
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     mobile_element  3866..4090
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     primer_bind     complement(4224..4241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4395..4983)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4475..4494)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5154..5945,5946..6014))
                     /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     5777..5796
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6015..6119)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(6145..6599)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6282..6303)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6493..6512
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     CDS             complement(join(6640..7319,1..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 tactgcagct tcacagcgga gttagcacgc agagcagtca cgatcagggg ccaagccagg
     1441 ttgggagagt tgtccatgga gatctcggac agctgcacga tcttggagtc agcgtccacc
     1501 acttgctgga tctcccacag agcggaagcg taggtgaagg tggtgccgtc gcaagtgttc
     1561 ttgtaggtgt tgtagtcggg gatgaccacc atcagcttag cagcggtggt cagagggatg
     1621 atgttcaggg gcacgcaacc gtcacgagcg ttgttgatga tgttgttcag agcgtcgttg
     1681 tccagcttgc ggagcatggt gaacagcatg gtctgcatag cggaggtcac tttagcacgc
     1741 ttgtcctcgg aacgagcctg cttgtacatc tgggtcatag cctggtcagc catcttctcg
     1801 agcttgcgct gcatagcagc gtcacggtcg aactcggact tggccacgtt cagggacttt
     1861 ttcagcttct tcagcaccac ctcagagtcg ccgttagcca cagcctgctc gtaagcctct
     1921 tgagcggtag cgaaagcagc gtaggagggc agagaggaga actcggaagc gatagcggag
     1981 ccaccagatc caccctgcag ggtagcacgg ttgtccagca tctcttcgca cagcttgttg
     2041 atgtccacag caccttgcat agacagcagg acggacagca gggacaccat cttctcgaaa
     2101 gcctcggtgg tgtccttagc cagcaggatg tcgttgtgca gttggacgca ctgagcccac
     2161 agcttgctgg aggattccac acgcagctgt tgcagcacag acagcagcac cacggaggtg
     2221 cacttcacgt cggacatctt ggacatggtg gctggatccg cgcccgatgg tgggacggta
     2281 tgaataatcc ggaatattta taggtttttt tattacaaaa ctgttacgaa aacagtaaaa
     2341 tacttattta tttgcgagat ggttatcatt ttaattatct ccatgatcta ttaatattcc
     2401 ggagtaggtc gatttaaatc tttagaccat agagcgttca cttgacagag tgccagccct
     2461 gggaccgaac cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt attctgtctt
     2521 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     2581 ctcgataaaa accccgattg agatatagtt taaacccaga taacttcgta taatgtatgc
     2641 tatacgaagt tatggtaccg cggccgcgta gaggatctgt tgatcagcag ttcaacctgt
     2701 tgatagtaca aacaaagctg tagccaacca ctagaactat agctagagtc ctgggcgaac
     2761 aaacgatgct cgccttccag aaaaccgagg atgcgaacca cttcatccgg ggtcagcacc
     2821 accggcaagc gccgcgacgg ccgaggtcta ccgatctcct gaagcaaggg cagatccgtg
     2881 cacagcacct tgccgtagaa gaacagcaag gccgccaatg cctgacgatg cgtggagacc
     2941 gaaaccttgc gctcgttcgc cagccaggac agaaatgcct cgacttcgct gctgcccaag
     3001 gttgccgggt gacgcacacc gtggaaacgg atgaaggcac aaacccagtt gacataagcc
     3061 tgttcggttc gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc
     3121 ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt
     3181 ttttttgtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga
     3241 tgtttgatgt tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg
     3301 cagtcgccct aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct
     3361 cggccctgac caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg
     3421 agacgtagcc acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg
     3481 tagtaagaca ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc
     3541 ggcttacgtt ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc
     3601 agtctccggc gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca
     3661 tgaggccaac gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc
     3721 cgcagtggct ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga
     3781 cccaagtacc gccacctaac aattcgttca agccgagatc ggcttcccgg ccgcggagtt
     3841 gttcggtaaa ttgtcacaac gccgcgaata tagtctttac catgcccttg gccacgcccc
     3901 tctttaatac gacgggcaat ttgcacttca gaaaatgaag agtttgcttt agccataaca
     3961 aaagtccagt atgctttttc acagcataac tggactgatt tcagtttaca actattctgt
     4021 ctagtttaag actttattgt catagtttag atctattttg ttcagtttaa gactttattg
     4081 tccgcccaca cccgcttacg cagggcatcc atttattact caaccgtaac cgattttgcc
     4141 aggttacgcg gctggtctat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     4201 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     4261 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     4321 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     4381 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     4441 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     4501 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     4561 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     4621 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     4681 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     4741 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     4801 ttgaagtggt ggcctaacta cggctacact agaagaatag tatttggtat ctgcgctctg
     4861 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     4921 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     4981 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     5041 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     5101 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     5161 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     5221 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     5281 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     5341 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     5401 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     5461 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     5521 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     5581 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     5641 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     5701 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     5761 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     5821 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     5881 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     5941 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     6001 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     6061 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     6121 acatttcccc gaaaagtgcc acctaaattg taagcgttaa tattttgtta aaattcgcgt
     6181 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     6241 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     6301 cactattaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg
     6361 cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact
     6421 aaatcggaac cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt
     6481 ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc
     6541 ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc
     6601 ccattcgcca atccgtcggc ttgaacgaat tgttagacat tatttgccga ctaccttggt
     6661 gatctcgcct ttcacgtagt ggacaaattc ttccaactga tctgcgcgcg aggccaagcg
     6721 atcttcttct tgtccaagat aagcctgtct agcttcaagt atgacgggct gatactgggc
     6781 cggcaggcgc tccattgccc agtcggcagc gacatccttc ggcgcgattt tgccggttac
     6841 tgcgctgtac caaatgcggg acaacgtaag cactacattt cgctcatcgc cagcccagtc
     6901 gggcggcgag ttccatagcg ttaaggtttc atttagcgcc tcaaatagat cctgttcagg
     6961 aaccggatca aagagttcct ccgccgctgg acctaccaag gcaacgctat gttctcttgc
     7021 ttttgtcagc aagatagcca gatcaatgtc gatcgtggct ggctcgaaga tacctgcaag
     7081 aatgtcattg cgctgccatt ctccaaattg cagttcgcgc ttagctggat aacgccacgg
     7141 aatgatgtcg tcgtgcacaa caatggtgac ttctacagcg cggagaatct cgctctctcc
     7201 aggggaagcc gaagtttcca aaaggtcgtt gatcaaagct cgccgcgttg tttcatcaag
     7261 ccttacggtc accgtaacca gcaaatcaat atcactgtgt ggcttcaggc cgccatcca
//
