LOCUS       sequence-435533-        8355 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In mammalian cells expresses a Quartet of receptor-binding domains 
            (RBDs) from the sarbecoviruses pang17, RmYN02, Rf1, and WIV1 each 
            separated by GS-linkers with an N-terminal SpyTag fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8355)
  AUTHORS   Hills RA, Tan TK, Cohen AA, Keeffe JR, Keeble AH, Gnanapragasam PNP,
            Storm KN, Rorick AV, West AP Jr, Hill ML, Liu S, Gilbert-Jaramillo 
            J, Afzal M, Napier A, Admans G, James WS, Bjorkman PJ, Townsend AR, 
            Howarth MR
  TITLE     Proactive vaccination using multiviral Quartet Nanocages to elicit 
            broad anti-coronavirus responses.
  JOURNAL   Nat Nanotechnol. 2024 May 6. doi: 10.1038/s41565-024-01655-9.
  PUBMED    38710880
REFERENCE   2  (bases 1 to 8355)
  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"; journalName: "Nat 
            Nanotechnol. 2024 May 6. doi: 10.1038/s41565-024-01655-9."
COMMENT     Sequence Label: SpyTag-Alternate Quartet
FEATURES             Location/Qualifiers
     source          1..8355
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1010..1048
                     /codon_start=1
                     /product="peptide derived from the CnaB2 adhesin domain of 
                     Streptococcus pyogenes FbaB"
                     /label=SpyTag
                     /note="forms a covalent isopeptide bond with the SpyCatcher
                     protein (Zakeri et al., 2012)"
                     /translation="AHIVMVDAYKPTK"
     primer_bind     complement(3946..3963)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3952..4176
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4222..4650
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4309..4328)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4519..4540
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4659..4679)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4664..4993
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4844..4979
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4906..4925
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5060..5854
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5114..5133)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5724..5743
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6028..6149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6065..6084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6119..6138
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6198..6214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6198..6214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6211..6233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6222..6238
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6246..6252,6253..6270,6271..6276))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6291..6312
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6429..6446)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6600..7188)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6680..6699)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7359..8150,8151..8219))
                     /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     7982..8001
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8220..8324)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccaccatg aacacccaga tcctggtgtt
      961 cgccctgatc gccatcattc ccaccaacgc cgacaagatc ggatctggcg cccacatcgt
     1021 gatggtggac gcctacaagc ctaccaaagg cagcggtgga agcggcggat ctggaaccgg
     1081 tcgggtgcag cccaccatca gcatcgtgcg gttccccaat atcaccaatc tgtgcccctt
     1141 cggcgaggtg ttcaacgcca gcaagtttgc ctctgtgtac gcctggaacc ggaagcggat
     1201 cagcaattgc gtggccgact acagcgtgct gtacaacagc accagcttca gcaccttcaa
     1261 gtgctacggc gtgtccccta ccaagctgaa cgacctgtgc ttcacaaacg tgtacgccga
     1321 cagcttcgtg gtcaagggcg acgaagttcg gcagattgcc cctggacaga ctggcgtgat
     1381 cgccgattac aactacaagc tgcccgacga cttcaccggc tgtgtgattg cctggaacag
     1441 cgtgaagcag gatgccctga caggcggcaa ttacggctac ctgtaccggc tgttcagaaa
     1501 gagcaagctg aagcccttcg agcgggacat ctccaccgag atctatcagg ccggcagcac
     1561 cccttgcaat ggccaagttg gcctgaactg ctactaccct ctggaaagat acggctttca
     1621 ccccaccacc ggcgtgaact accagccatt cagagtggtg gtgctgagct tcgagctgct
     1681 gaatggacct gccacagtgt gcggccctaa gctgtctaca accctggtca aggataagtg
     1741 cgtgaacttc ggctcaggtg gatccggagg tacgggaaga atcctgccta gcacagaggt
     1801 cgtgcggttc cccaacatca ccaatttctg ccccttcgac aaggtgttca acgccaccag
     1861 atttcccaac gtgtacgcct ggcagcggac caagatcagc gactgtatcg ccgactacac
     1921 cgtgctgtac aacagcacca gcttcagcac cttcaagtgc tacggcgtgt cccctagcaa
     1981 gctgatcgac ctgtgcttca cctctgtgta cgccgacacc ttcctgatcc ggttctctga
     2041 agtgcggcag atcgcccctg gcgaaacagg cgtgatcgcc gattacaact acaagctgcc
     2101 cgacgacttc accggctgtg tgctggcttg gaatactgcc cagcaggaca tcggcagcta
     2161 cttctacaga tcccaccggg ccgtgaagct gaagcccttc gagagagatc tgagcagcga
     2221 cgagaatggc gtgcggaccc tgagcaccta cgacttcaac cccaatgtgc ccctggacta
     2281 ccaggccaca agagtggtgg tgctgagctt cgagctgctg aatgcccctg ccacagtgtg
     2341 tggccctaag ctgtctaccc agctggtcaa gaacagatgc gtgaacttcg ggtccggtgg
     2401 gtcgggcggt agtgggagag tgtctcctgt gacagaggtc gtgcggttcc ccaacatcac
     2461 caatctgtgc cccttcgaca aggtgttcaa cgccaccaga tttcccagcg tgtacgcctg
     2521 ggagcgcacc aagatctctg attgcgtggc cgactacacc gtgttctaca acagcaccag
     2581 cttcagcacc ttcaactgct acggcgtgtc ccctagcaag ctgatcgacc tgtgcttcac
     2641 ctctgtgtac gccgacacct tcctgatccg gttcagcgaa gtgcgacagg tggcacctgg
     2701 acagacaggc gtgatcgccg attacaacta caagctgccc gacgacttca ccggctgtgt
     2761 gatcgcctgg aataccgcca agcaggacgt gggcagctac ttctacagaa gccacagaag
     2821 ctccaagctg aagcccttcg agagggacct gagcagcgaa gaaaatggcg tgcggaccct
     2881 gagcacctac gacttcaacc agaacgtgcc cctggaatat caggccacca gagtggtggt
     2941 gctgagcttc gagctgctga atgcccctgc cacagtgtgt ggccctaagc tgtctaccag
     3001 cctggtcaag aaccagtgcg tgaacttcgg tagcgggggg tcggggggca ccggcagagt
     3061 ggcccctagc aaagaagtcg tgcggttccc caatatcacc aatctgtgcc ccttcggcga
     3121 ggtgttcaac gccaccacat ttccctctgt gtacgcctgg gagagaaagc ggatcagcaa
     3181 ctgcgtggcc gactacagcg tgctgtacaa cagcacctcc tttagcacct tcaagtgcta
     3241 cggcgtgtcc gccaccaagc tgaacgatct gtgcttctcc aacgtgtacg ccgacagctt
     3301 cgtggtcaag ggcgacgatg tgcggcagat tgctcctgga cagacaggcg tgatcgccga
     3361 ttacaactac aagctgcctg acgacttcac cggctgtgtg ctggcctgga acaccagaaa
     3421 catcgacgcc acacagaccg gcaactacaa ttacaagtac agaagcctgc ggcacggcaa
     3481 gctgaggcct ttcgagaggg acatcagcaa tgtgcctttc agccccgacg gcaagccttg
     3541 tacacctcct gctttcaact gctactggcc cctgaacgac tacggcttct acatcacaaa
     3601 cggcatcggc tatcagccct acagagtggt ggtgctgtcc ttcgagctgc tgaatgcccc
     3661 tgccacagtg tgtggcccta agctgtctac cgacctgatc aagaaccagt gcgtgaactt
     3721 ctagtagacc ggtaagctga gcagcggcta caaggacgtg atcctgtggt tcagcttcgg
     3781 cgccagctgc ttcatcctgc tggccatcgt gatgggcctg gtgttcatct gcgtgaagaa
     3841 cggcaacatg cggtgcacca tctgcatctg atgatatcga attccaccac actggactag
     3901 tggatccgag ctcggtacca agcttaagtt taaaccgctg atcagcctcg actgtgcctt
     3961 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     4021 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     4081 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     4141 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     4201 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     4261 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4321 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc
     4381 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4441 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4501 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4561 cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg
     4621 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4681 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4741 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     4801 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     4861 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     4921 cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     4981 aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac
     5041 aggatgagga tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc
     5101 ttgggtggag aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc
     5161 cgccgtgttc cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc
     5221 cggtgccctg aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg
     5281 cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt
     5341 gggcgaagtg ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc
     5401 catcatggct gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga
     5461 ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga
     5521 tcaggatgat ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct
     5581 caaggcgcgc atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc
     5641 gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt
     5701 ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg
     5761 cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat
     5821 cgccttctat cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc
     5881 gaccaagcga cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa
     5941 aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat
     6001 ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa
     6061 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     6121 ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag
     6181 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6241 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6301 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6361 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6421 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6481 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6541 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6601 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6661 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6721 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     6781 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     6841 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     6901 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     6961 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7021 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7081 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7141 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7201 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7261 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7321 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7381 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7441 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7501 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7561 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7621 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7681 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7741 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     7801 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     7861 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     7921 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     7981 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8041 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8101 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8161 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     8221 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     8281 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     8341 gtgccacctg acgtc
//
