LOCUS       sequence-333208-        6804 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Baculoviral transfer vector to express 3xFlag-His6-nsp13 
            (SARS-CoV-2) in insect cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6804)
  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 6804)
  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_3xFlag-His6-nsp13 (SARS-CoV-2)
FEATURES             Location/Qualifiers
     source          1..6804
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(join(90..95,96..112,113..118))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    complement(514..547)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     promoter        595..686
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     611..628
                     /label=Polyhedrin-F
                     /note="Polyhedrin promoter, forward primer"
     regulatory      730..739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             739..804
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             811..828
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    2779..2913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2816..2835)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2870..2889
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     mobile_element  3023..3188
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3372..3826
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3459..3478)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3668..3689
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3852..3956
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3957..4025,4026..4817)
                     /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(4175..4194)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4988..5576
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5477..5496
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5730..5747
                     /label=L4440
                     /note="L4440 vector, forward primer"
     mobile_element  5881..6105
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6172..6705)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 ccatgaaaac cgccactgcg ccgttaccac cgctgcgttc ggtcaaggtt ctggaccagt
       61 tgcgtgagcg catacgctac ttgcattaca gtttacgaac cgaacaggct tatgtcaact
      121 gggtttgtgc cttcatccgt ttccacggtg tgcgtcaccc ggcaaccttg ggcagcagcg
      181 aagtcgaggc atttctgtcc tggctggcga acgagcgcaa ggtttcggtc tccacgcatc
      241 gtcaggcatt ggcggccttg ctgttcttct acggcaaggt gctgtgcacg gatctgccct
      301 tgcttcagga gatcggtaga cctcggccgt cgcggcgctt gccggtggtg ctgaccccgg
      361 atgaagtggt tcgcatcctc ggttttctgg aaggcgagca tcgtttgttc gcccaggact
      421 ctagctatag ttctagtggt tggctacagc tttgtttgta ctatcaacag gttgaactgc
      481 tgatcaacag atcctctacg cggccgcggt accataactt cgtatagcat acattatacg
      541 aagttatctg gttttactag tatcgattcg cgacctactc cggaatatta atagatcatg
      601 gagataatta aaatgataac catctcgcaa ataaataagt attttactgt tttcgtaaca
      661 gttttgtaat aaaaaaacct ataaatattc cggattattc ataccgtccc accatcgggc
      721 gcggatccag ccaccatgga ctacaaggac cacgacggcg attacaagga tcatgacatc
      781 gactacaaag acgacgatga caagggttcc caccatcacc accatcactc tgctgtgctg
      841 caggctgttg gtgcttgcgt gctgtgcaac tcccagacct ctttgcgttg cggtgcttgt
      901 atccgtcgtc cattcctgtg ctgcaagtgc tgctacgacc acgtgatctc cacctctcac
      961 aagctggtgc tgtctgtgaa cccctacgtg tgcaacgctc ctggttgcga cgtgaccgat
     1021 gtgacccaac tgtacctcgg tggcatgtcc tactactgca agtcccacaa gcctccaatc
     1081 agcttccctc tgtgcgctaa cggacaggtg ttcggcctgt acaagaacac ttgcgtgggt
     1141 tccgacaacg tcaccgactt caacgctatc gctacctgcg actggaccaa cgctggcgac
     1201 tacatcctgg ctaacacttg caccgagcgt ctgaagctgt tcgctgctga gactctgaag
     1261 gctaccgagg aaaccttcaa gctgtcctac ggtatcgcca ccgtgcgtga agtgctgtcc
     1321 gaccgtgaac tgcacctgtc ctgggaagtg ggaaagcctc gtcctcctct gaaccgtaac
     1381 tacgtgttca ccggttaccg tgtgaccaag aactccaagg tgcagatcgg cgagtacacc
     1441 ttcgagaagg gcgattacgg cgacgctgtg gtgtaccgtg gtactaccac ctacaagctg
     1501 aacgtgggcg actacttcgt gctgacttcc cacaccgtga tgcccctgtc tgctcctact
     1561 ctggtgcccc aagagcacta cgtgcgtatc accggactgt accccactct gaatatctcc
     1621 gacgagttct cctccaacgt ggccaactac cagaaagtgg gcatgcagaa gtactccaca
     1681 ctgcagggtc cacctggcac cggaaagtcc cacttcgcta tcggactggc tctgtactac
     1741 ccctccgctc gtatcgtgta caccgcttgc tctcacgctg ctgtggacgc tctgtgcgag
     1801 aaggctctga agtacctgcc tatcgacaag tgctcccgta tcatccccgc tcgtgctcgt
     1861 gtggaatgct tcgacaagtt caaagtgaac agcaccctcg agcagtacgt gttctgcacc
     1921 gtgaacgctc tgcccgagac taccgctgac atcgtggtgt tcgacgagat ctccatggct
     1981 accaactacg acctgtctgt ggtcaacgct cgtctgcgtg ctaaacacta cgtgtacatc
     2041 ggcgaccccg ctcagttgcc tgctcctcgt actctgctga ccaagggcac tctggaaccc
     2101 gagtacttca actccgtgtg ccgtctgatg aagactatcg gtcccgacat gttcctggga
     2161 acctgcagac gttgccctgc tgagatcgtg gacaccgtgt ctgctctggt gtacgacaac
     2221 aagctcaagg ctcacaagga caagtccgct cagtgcttca agatgttcta caagggtgtc
     2281 atcacccacg acgtgtcctc cgctatcaac cgtcctcaga tcggtgtcgt gcgcgagttc
     2341 ctgactcgta atcctgcttg gcgcaaggct gtgttcatct ccccttacaa ctcccaaaac
     2401 gctgtggctt ccaagatcct gggcctgcct actcagaccg tggactcctc tcagggttcc
     2461 gagtacgact acgtcatctt cacccagact accgagactg ctcactcctg caacgtgaac
     2521 cgtttcaacg tcgccatcac cagggctaaa gtcggtatcc tgtgcatcat gtccgacagg
     2581 gacctctacg acaagctcca gttcacctcc ttggagatcc ctcgtcgtaa cgtggcgact
     2641 ctgcagtaaa agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta
     2701 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     2761 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2821 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2881 aaactcatca atgtatctta tcatgtctgg atctgatcac tgcttgagcc tagaagatcc
     2941 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
     3001 atcataaccc ctaggagatc cgaaccagat aagtgaaatc tagttccaaa ctattttgtc
     3061 atttttaatt ttcgtattag cttacgacgc tacacccagt tcccatctat tttgtcactc
     3121 ttccctaaat aatccttaaa aactccattt ccacccctcc cagttcccaa ctattttgtc
     3181 cgcccacagc ggggcatttt tcttcctgtt atgtttttaa tcaaacatcc tgccaactcc
     3241 atgtgacaaa ccgtcatctt cggctacttt ttctctgtca cagaatgaaa atttttctgt
     3301 catctcttcg ttattaatgt ttgtaattga ctgaatatca acgcttattt gcagcctgaa
     3361 tggcgaatgg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3421 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3481 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3541 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3601 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3661 cttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct
     3721 tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa
     3781 caaaaattta acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc
     3841 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     3901 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     3961 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     4021 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     4081 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     4141 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     4201 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     4261 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     4321 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     4381 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     4441 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     4501 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     4561 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     4621 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     4681 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     4741 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     4801 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     4861 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4921 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4981 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     5041 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     5101 ctggcttcag cagagcgcag ataccaaata ctattcttct agtgtagccg tagttaggcc
     5161 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     5221 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     5281 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     5341 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     5401 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     5461 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     5521 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     5581 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     5641 ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata
     5701 ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc
     5761 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagaccagc
     5821 cgcgtaacct ggcaaaatcg gttacggttg agtaataaat ggatgccctg cgtaagcggg
     5881 tgtgggcgga caataaagtc ttaaactgaa caaaatagat ctaaactatg acaataaagt
     5941 cttaaactag acagaatagt tgtaaactga aatcagtcca gttatgctgt gaaaaagcat
     6001 actggacttt tgttatggct aaagcaaact cttcattttc tgaagtgcaa attgcccgtc
     6061 gtattaaaga ggggcgtggc caagggcatg gtaaagacta tattcgcggc gttgtgacaa
     6121 tttaccgaac aactccgcgg ccgggaagcc gatctcggct tgaacgaatt gttaggtggc
     6181 ggtacttggg tcgatatcaa agtgcatcac ttcttcccgt atgcccaact ttgtatagag
     6241 agccactgcg ggatcgtcac cgtaatctgc ttgcacgtag atcacataag caccaagcgc
     6301 gttggcctca tgcttgagga gattgatgag cgcggtggca atgccctgcc tccggtgctc
     6361 gccggagact gcgagatcat agatatagat ctcactacgc ggctgctcaa acttgggcag
     6421 aacgtaagcc gcgagagcgc caacaaccgc ttcttggtcg aaggcagcaa gcgcgatgaa
     6481 tgtcttacta cggagcaagt tcccgaggta atcggagtcc ggctgatgtt gggagtaggt
     6541 ggctacgtct ccgaactcac gaccgaaaag atcaagagca gcccgcatgg atttgacttg
     6601 gtcagggccg agcctacatg tgcgaatgat gcccatactt gagccaccta actttgtttt
     6661 agggcgactg ccctgctgcg taacatcgtt gctgctgcgt aacatcgttg ctgctccata
     6721 acatcaaaca tcgacccacg gcgtaacgcg cttgctgctt ggatgcccga ggcatagact
     6781 gtacaaaaaa acagtcataa caag
//
