LOCUS Exported 7374 bp ds-DNA circular SYN 19-MAY-2021 DEFINITION Baculoviral transfer vector to express GST-nsp13 (SARS-CoV-2) in insect cells. ACCESSION . VERSION . KEYWORDS pLIB_GST-nsp13 (SARS-CoV-2) 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, Kjae 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 bioRxiv https://www.biorxiv.org/content/10.1101/2021.04.07.438808v1 REFERENCE 2 (bases 1 to 7374) AUTHORS . TITLE Direct Submission JOURNAL Exported May 19, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7374 /organism="synthetic DNA construct" /mol_type="other DNA" protein_bind 414..447 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." 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" /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 4427..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" /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(7364..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 //