LOCUS       sequence-435535-        8364 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In mammalian cells expresses a Quartet of receptor-binding domains 
            (RBDs) from the sarbecoviruses SHC014, Rs4081, RaTG13, and 
            SARS-CoV-2 XBB.1.5 separated by GS-linkers with an N-terminal 
            SpyTag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8364)
  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 8364)
  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-Kraken Quartet
FEATURES             Location/Qualifiers
     source          1..8364
                     /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(3955..3972)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3961..4185
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4231..4659
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4318..4337)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4528..4549
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4668..4688)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4673..5002
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4853..4988
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4915..4934
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5069..5863
                     /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(5123..5142)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5733..5752
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6037..6158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6074..6093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6128..6147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6207..6223)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6207..6223)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6220..6242)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6231..6247
                     /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(6255..6261,6262..6279,6280..6285))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6300..6321
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6438..6455)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6609..7197)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6689..6708)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7368..8159,8160..8228))
                     /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     7991..8010
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8229..8333)
                     /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 tagagtggcc cctagcaaag aagtcgtgcg gttccccaat atcaccaatc tgtgcccctt
     1141 cggcgaggtg ttcaacgcca ccacatttcc ctctgtgtac gcctgggaga gaaagcggat
     1201 cagcaactgc gtggccgact acagcgtgct gtacaacagc acctccttta gcaccttcaa
     1261 gtgctacggc gtgtccgcca ccaagctgaa cgatctgtgc ttctccaacg tgtacgccga
     1321 cagcttcgtg gtcaagggcg acgatgtgcg gcagattgct cctggacaga caggcgtgat
     1381 cgccgattac aactacaagc tgcctgacga cttcctgggc tgtgtgctgg cctggaacac
     1441 caatagcaag gacagctcca ccagcggcaa ctacaactac ctgtacagat gggtccgacg
     1501 cagcaagctg aatccctacg agagggacct gagcaacgac atctatagcc ctggcggcca
     1561 gagctgttct gccgtgggac ccaattgcta caaccctctg aggccctacg gcttctttac
     1621 cacagctggc gtgggccacc agccttacag agtggtggtg ctgtcattcg agctgctgaa
     1681 tgcccctgcc acagtgtgtg gccctaagct gtctaccgac ctgatcaaga accaggggag
     1741 tggtgggtcc gggggttcgg gacgggtgtc ccctacacac gaggtcgtgc ggttccccaa
     1801 tatcaccaac agatgcccct tcgacaaggt gttcaacgcc agcagattcc ccaacgtgta
     1861 cgcctgggag cgcaccaaga tcagcgattg cgtggccgat tacaccgtgc tgtacaactc
     1921 caccagcttt agcaccttta agtgctacgg cgtgtcccca agcaagctga tcgacctgtg
     1981 cttcacctct gtgtacgccg acaccttcct gatccggtct agcgaagtgc gacaagtggc
     2041 ccctggcgaa acaggcgtga tcgccgacta caactacaag ctgcccgacg acttcaccgg
     2101 ctgtgtgatc gcctggaata ccgccaagca ggaccagggc cagtactact acagaagcag
     2161 cagaaagacc aagctcaagc ccttcgagcg ggacctgacc tccgatgaga atggcgtcag
     2221 aaccctgtcc acctacgatt tctaccccaa tgtgcccatc gagtaccagg ccaccagagt
     2281 ggtggtgctg tctttcgaac tgctgaacgc ccctgccaca gtgtgtggcc ctaaactgtc
     2341 tacagccctg gtcaagaacc agtgcgtgaa cttcggcagc ggcggaacgg gaggtagtgg
     2401 ccgggtgcag cccaccgata gcatcgtgcg gttccccaat atcaccaatc tgtgcccctt
     2461 cggcgaggtg ttcaatgcca ccacattcgc ctctgtgtac gcctggaacc ggaagcggat
     2521 cagcaattgc gtggccgact actccgtgct gtacaacagc accagcttca gcaccttcaa
     2581 gtgctacggc gtgtccccta ccaagctgaa cgacctgtgc ttcacaaacg tgtacgccga
     2641 cagcttcgtg atcaccgggg atgaagtgcg gcagattgcc cctggacaga caggcaagat
     2701 cgccgactac aactacaagc tgcccgacga cttcaccggc tgtgtgattg cctggaactc
     2761 caagcacatc gacgccaaag aaggcgggaa cttcaactac ctgtaccggc tgttccggaa
     2821 ggccaacctg aagcctttcg agcgggatat ctccaccgag atctaccagg ccggcagcaa
     2881 gccttgtaat ggccagacag gcctgaactg ctactaccca ctgtacagat acggcttcta
     2941 ccccacagac ggcgtgggcc accagcctta tagagtggtg gtgctgagct tcgagctgct
     3001 gaatgcccct gccacagtgt gcggccctaa gaaaagcacc aatctcgtga agaacaaatg
     3061 cgtgaacttc ggtacgggtg gcagtggagg gagcgggagg gtgcagccca ccgagagcat
     3121 cgtgaggttc cccaatataa cgaatctgtg ccccttccac gaggtcttta acgccaccac
     3181 attcgcgagt gtttacgcgt ggaacagaaa gcggatatcc aactgtgttg ccgattatag
     3241 cgtcatctat aattttgccc cgttctttgc ctttaagtgt tatggggtga gtccgacgaa
     3301 attgaacgat ctttgcttca cgaacgttta tgcggactcc ttcgtaatac gaggcaatga
     3361 ggtaagtcag atagctcccg gacagactgg gaatattgcg gactacaatt acaaacttcc
     3421 agacgatttt accggatgcg taattgcctg gaacagtaat aaactggata gtaaacctag
     3481 tggtaattac aattatctct acagattgtt taggaaaagc aagcttaaac cgttcgaaag
     3541 agacatatcc acggaaatat atcaggccgg gaacaaacca tgcaatgggg ttgccggccc
     3601 aaactgttat agcccactgc aatcatacgg ctttcgccct acatatggcg tcgggcacca
     3661 gccgtacaga gtggtagttc tcagcttcga gctccttcac gcaccggcta cggtgtgcgg
     3721 accgaaaaag tagtagaccg gtaagctgag cagcggctac aaggacgtga tcctgtggtt
     3781 cagcttcggc gccagctgct tcatcctgct ggccatcgtg atgggcctgg tgttcatctg
     3841 cgtgaagaac ggcaacatgc ggtgcaccat ctgcatctga tgatatcgaa ttccaccaca
     3901 ctggactagt ggatccgagc tcggtaccaa gcttaagttt aaaccgctga tcagcctcga
     3961 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4021 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4081 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     4141 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     4201 gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg
     4261 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     4321 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     4381 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     4441 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     4501 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     4561 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     4621 taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca
     4681 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4741 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4801 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4861 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4921 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     4981 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     5041 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     5101 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     5161 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     5221 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     5281 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     5341 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     5401 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     5461 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     5521 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     5581 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     5641 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     5701 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     5761 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     5821 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     5881 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     5941 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     6001 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     6061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     6121 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     6181 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     6241 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     6301 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     6361 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     6421 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     6481 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     6541 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     6601 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     6661 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     6721 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     6781 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     6841 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     6901 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     6961 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     7021 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     7081 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     7141 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     7201 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     7261 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     7321 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     7381 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     7441 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     7501 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     7561 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     7621 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     7681 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     7741 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     7801 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     7861 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     7921 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     7981 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     8041 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     8101 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     8161 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     8221 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     8281 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     8341 ccccgaaaag tgccacctga cgtc
//
