LOCUS       sequence-333224-        7374 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculoviral transfer vector to express GST-nsp13 (SARS-CoV-2) in 
            insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  AUTHORS   Zeng J, Weissmann F, Bertolin AP, Posse V, Canal B, Ulferts R, Wu M,
            Harvey R, Hussain S, Milligan JC, Roustan C, Borg A, McCoy L, Drury 
            LS, Kjaer S, McCauley J, Howell M, Beale R, Diffley JFX
  TITLE     Identifying SARS-CoV-2 antiviral compounds by screening for small 
            molecule inhibitors of nsp13 helicase.
  JOURNAL   Biochem J. 2021 Jul 16;478(13):2405-2423. doi: 10.1042/BCJ20210201.
  PUBMED    34198322
REFERENCE   2  (bases 1 to 7374)
  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: "2405-2423"
COMMENT     Sequence Label: pLIB_GST-nsp13 (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..7374
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    complement(414..447)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        495..586
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     511..528
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     CDS             636..1289
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     1247..1269
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             1296..1313
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     polyA_signal    3249..3383
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3286..3305)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3340..3359
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3493..3658
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3842..4296
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3929..3948)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4138..4159
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4322..4426
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4427..4495,4496..5287)
                     /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(4645..4664)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5458..6046
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5947..5966
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6200..6217
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  6351..6575
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6642..7175)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(7364..7369,7370..7374,1..12,13..18))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
ORIGIN
        1 cgaacaggct tatgtcaact gggtttgtgc cttcatccgt ttccacggtg tgcgtcaccc
       61 ggcaaccttg ggcagcagcg aagtcgaggc atttctgtcc tggctggcga acgagcgcaa
      121 ggtttcggtc tccacgcatc gtcaggcatt ggcggccttg ctgttcttct acggcaaggt
      181 gctgtgcacg gatctgccct tgcttcagga gatcggtaga cctcggccgt cgcggcgctt
      241 gccggtggtg ctgaccccgg atgaagtggt tcgcatcctc ggttttctgg aaggcgagca
      301 tcgtttgttc gcccaggact ctagctatag ttctagtggt tggctacagc tttgtttgta
      361 ctatcaacag gttgaactgc tgatcaacag atcctctacg cggccgcggt accataactt
      421 cgtatagcat acattatacg aagttatctg gttttactag tatcgattcg cgacctactc
      481 cggaatatta atagatcatg gagataatta aaatgataac catctcgcaa ataaataagt
      541 attttactgt tttcgtaaca gttttgtaat aaaaaaacct ataaatattc cggattattc
      601 ataccgtccc accatcgggc gcggatccag ccaccatgtc ccctatacta ggttattgga
      661 aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg
      721 aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg
      781 gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta
      841 tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc
      901 gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa
      961 ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa
     1021 tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa
     1081 cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt
     1141 gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa
     1201 ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca
     1261 cgtttggtgg tggcgaccat cctccaaaat cggatctggt tccgcgtgga tccgctgttg
     1321 gtgcttgcgt gctgtgcaac tcccagacct ctttgcgttg cggtgcttgt atccgtcgtc
     1381 cattcctgtg ctgcaagtgc tgctacgacc acgtgatctc cacctctcac aagctggtgc
     1441 tgtctgtgaa cccctacgtg tgcaacgctc ctggttgcga cgtgaccgat gtgacccaac
     1501 tgtacctcgg tggcatgtcc tactactgca agtcccacaa gcctccaatc agcttccctc
     1561 tgtgcgctaa cggacaggtg ttcggcctgt acaagaacac ttgcgtgggt tccgacaacg
     1621 tcaccgactt caacgctatc gctacctgcg actggaccaa cgctggcgac tacatcctgg
     1681 ctaacacttg caccgagcgt ctgaagctgt tcgctgctga gactctgaag gctaccgagg
     1741 aaaccttcaa gctgtcctac ggtatcgcca ccgtgcgtga agtgctgtcc gaccgtgaac
     1801 tgcacctgtc ctgggaagtg ggaaagcctc gtcctcctct gaaccgtaac tacgtgttca
     1861 ccggttaccg tgtgaccaag aactccaagg tgcagatcgg cgagtacacc ttcgagaagg
     1921 gcgattacgg cgacgctgtg gtgtaccgtg gtactaccac ctacaagctg aacgtgggcg
     1981 actacttcgt gctgacttcc cacaccgtga tgcccctgtc tgctcctact ctggtgcccc
     2041 aagagcacta cgtgcgtatc accggactgt accccactct gaatatctcc gacgagttct
     2101 cctccaacgt ggccaactac cagaaagtgg gcatgcagaa gtactccaca ctgcagggtc
     2161 cacctggcac cggaaagtcc cacttcgcta tcggactggc tctgtactac ccctccgctc
     2221 gtatcgtgta caccgcttgc tctcacgctg ctgtggacgc tctgtgcgag aaggctctga
     2281 agtacctgcc tatcgacaag tgctcccgta tcatccccgc tcgtgctcgt gtggaatgct
     2341 tcgacaagtt caaagtgaac agcaccctcg agcagtacgt gttctgcacc gtgaacgctc
     2401 tgcccgagac taccgctgac atcgtggtgt tcgacgagat ctccatggct accaactacg
     2461 acctgtctgt ggtcaacgct cgtctgcgtg ctaaacacta cgtgtacatc ggcgaccccg
     2521 ctcagttgcc tgctcctcgt actctgctga ccaagggcac tctggaaccc gagtacttca
     2581 actccgtgtg ccgtctgatg aagactatcg gtcccgacat gttcctggga acctgcagac
     2641 gttgccctgc tgagatcgtg gacaccgtgt ctgctctggt gtacgacaac aagctcaagg
     2701 ctcacaagga caagtccgct cagtgcttca agatgttcta caagggtgtc atcacccacg
     2761 acgtgtcctc cgctatcaac cgtcctcaga tcggtgtcgt gcgcgagttc ctgactcgta
     2821 atcctgcttg gcgcaaggct gtgttcatct ccccttacaa ctcccaaaac gctgtggctt
     2881 ccaagatcct gggcctgcct actcagaccg tggactcctc tcagggttcc gagtacgact
     2941 acgtcatctt cacccagact accgagactg ctcactcctg caacgtgaac cgtttcaacg
     3001 tcgccatcac cagggctaaa gtcggtatcc tgtgcatcat gtccgacagg gacctctacg
     3061 acaagctcca gttcacctcc ttggagatcc ctcgtcgtaa cgtggcgact ctgcagtaaa
     3121 agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac
     3181 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     3241 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     3301 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3361 atgtatctta tcatgtctgg atctgatcac tgcttgagcc tagaagatcc ggctgctaac
     3421 aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact atcataaccc
     3481 ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc atttttaatt
     3541 ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc ttccctaaat
     3601 aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc cgcccacagc
     3661 ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc atgtgacaaa
     3721 ccgtcatctt cggctacttt ttctctgtca cagaatgaaa atttttctgt catctcttcg
     3781 ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa tggcgaatgg
     3841 gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     3901 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     3961 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     4021 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     4081 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt cttaatagtg
     4141 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4201 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4261 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     4321 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     4381 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     4441 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     4501 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     4561 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     4621 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     4681 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     4741 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     4801 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     4861 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     4921 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     4981 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5041 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5101 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5161 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     5221 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     5281 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     5341 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5401 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5461 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5521 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5581 cagagcgcag ataccaaata ctattcttct agtgtagccg tagttaggcc accacttcaa
     5641 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5701 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5761 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5821 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5881 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5941 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6001 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     6061 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6121 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     6181 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     6241 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc cgcgtaacct
     6301 ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg tgtgggcgga
     6361 caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt cttaaactag
     6421 acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat actggacttt
     6481 tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc gtattaaaga
     6541 ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa tttaccgaac
     6601 aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc ggtacttggg
     6661 tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag agccactgcg
     6721 ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc gttggcctca
     6781 tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc gccggagact
     6841 gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag aacgtaagcc
     6901 gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa tgtcttacta
     6961 cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt ggctacgtct
     7021 ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg gtcagggccg
     7081 agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt agggcgactg
     7141 ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata acatcaaaca
     7201 tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact gtacaaaaaa
     7261 acagtcataa caagccatga aaaccgccac tgcgccgtta ccaccgctgc gttcggtcaa
     7321 ggttctggac cagttgcgtg agcgcatacg ctacttgcat tacagtttac gaac
//
