LOCUS       sequence-435534-        8397 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 
            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 8397)
  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 8397)
  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-Quartet [SARS1]
FEATURES             Location/Qualifiers
     source          1..8397
                     /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(3988..4005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3994..4218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4264..4692
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4351..4370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4561..4582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4701..4721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4706..5035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4886..5021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4948..4967
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5102..5896
                     /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(5156..5175)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5766..5785
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6070..6191
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6107..6126)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6161..6180
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6240..6256)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6240..6256)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6253..6275)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6264..6280
                     /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(6288..6294,6295..6312,6313..6318))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6333..6354
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6471..6488)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6642..7230)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6722..6741)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7401..8192,8193..8261))
                     /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     8024..8043
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8262..8366)
                     /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 gagcgggcgc gtggtcccgt ctggcgacgt
     3121 agtgagattt cctaatataa ccaatttgtg ccccttcggg gaagtattta acgcaacgaa
     3181 attcccgtcc gtctacgcgt gggaaaggaa gaagataagt aattgtgtag ccgattattc
     3241 tgtcttgtat aattcaacat tcttctcaac cttcaagtgt tacggggtct ccgcaacaaa
     3301 actcaatgat ctctgcttct caaacgtgta tgccgattct tttgtcgtaa agggagatga
     3361 tgtcagacaa atcgcgcccg ggcaaaccgg ggttatcgca gattataact ataaattgcc
     3421 tgacgacttc atgggctgtg tacttgcctg gaacacccga aatatcgatg ccacaagtac
     3481 cgggaactac aattacaagt acagatatct gaggcatggg aaattgcgcc cttttgagcg
     3541 agatatatca aatgttccct ttagccctga tggcaaacca tgtacacccc cagcattgaa
     3601 ctgctattgg cctcttaacg attacggttt ctacacaacc actggcattg gatatcagcc
     3661 ttaccgtgtc gttgtccttt ctttcgaact cttgaacgct ccggctaccg tgtgtggacc
     3721 taaactttcc accgatctga ttaagaacca atgtgtgaac ttttagtaga ccggtaagct
     3781 gagcagcggc tacaaggacg tgatcctgtg gttcagcttc ggcgccagct gcttcatcct
     3841 gctggccatc gtgatgggcc tggtgttcat ctgcgtgaag aacggcaaca tgcggtgcac
     3901 catctgcatc tgatgatatc gaattccacc acactggact agtggatccg agctcggtac
     3961 caagcttaag tttaaaccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     4021 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     4081 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     4141 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     4201 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     4261 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     4321 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     4381 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     4441 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     4501 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     4561 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     4621 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     4681 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     4741 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     4801 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     4861 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     4921 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     4981 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     5041 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     5101 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5161 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5221 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5281 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5341 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5401 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5461 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5521 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5581 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5641 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5701 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5761 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5821 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5881 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5941 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6001 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6061 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6121 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6181 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6241 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6301 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6361 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6421 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6481 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6541 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6601 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6661 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6721 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6781 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6841 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6901 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6961 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     7021 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     7081 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7141 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7201 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7261 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7321 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7381 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7441 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7501 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7561 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7621 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7681 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7741 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7801 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7861 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7921 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7981 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     8041 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     8101 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8161 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8221 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8281 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8341 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtc
//
